Biomedical

Second-harmonic generation (SHG) and two-photon excitation fluorescence (TPEF) are relatively new and promising tools for the detailed imaging of biological samples and processes at the microscopic level. By exploiting these nonlinear phenomena phototoxicity and photobleaching effects on the specimens are reduced dramatically. The main target of this work was the development of a compact inexpensive and reliable experimental apparatus for nonlinear microscopy measurements. Femtosecond laser pulses were utilized for excitation. We achieved high-resolution imaging and mapping of Caenorhabditis elegans (C. elegans) neurons and muscular structures of the pharynx, at the microscopic level by performing SHG and TPEF measurements. By detecting nonlinear phenomena such as SHG and TPEF it is feasible to extract valuable information concerning the structure and the function of nematode neurons.

In this work we describe the identification of the biologically active triterpenes and sterols present in the hexane extracts of six species of Miconia using gas chromatography. The main compounds present in these extracts are β-amyrin, α-amyrin, and β-sitosterol. The technique employed herein is shown to be a valuable and rapid tool for determining biologically active triterpenes and sterols present in non-polar extracts. EXPERIMENTAL Plant materials. Miconia species (M. albicans, M. pepericarpa, M. sellowiana, M. falax, M. rubiginosa and M. ligustroides) were collected along the Franca-Claraval highway in São Paulo, Brazil, and they were identified by

Abstract: Many dental clinical implant studies have fo- cused on the success of endosseous implants with a variety of surface characteristics. Most of the surface alterations have been aimed at achieving greater bone-to-implant con- tact as determined histometrically at the light microscopic ...

We determine temperature effect on the absorption and reduced scattering coefficients (a and s Ј) of human forearm skin. Optical and thermal simulation data suggest that a and s Ј are determined within a temperature-controlled depth of Ϸ2 mm. Cutaneous s Ј change linearly with temperature. Change in a was complex and irreversible above body normal temperatures. Light penetration depth (␦) in skin increased on cooling, with considerable person-toperson variations. We attribute the effect of temperature on s Ј to change in refractive index mismatch, and its effect on a to perfusion changes. The reversible temperature effect on s Ј was maintained during more than 90 min. contact between skin and the measuring probe, where temperature was modulated between 38 and 22°C for multiple cycles While temperature modulated s Ј instantaneously and reversibly, a exhibited slower response time and consistent drift. There was a statistically significant upward drift in a and a mostly downward drift in s Ј over the contact period. The drift in temperature-induced fractional change in s Ј was less statistically significant than the drift in s Ј. ⌬ s Ј values determined under temperature modulation conditions may have less nonspecific drift than s Ј which may have significance for noninvasive determination of analytes in human tissue.

The clinical performance of light polymerised dental composites is greatly in#uenced by the quality of the light-curing unit (LCU) used. Commonly used halogen LCUs have some speci"c drawbacks such as decreasing of the light output with time. This may result in low degree of monomer conversion of the composites with negative clinical implications. Previous studies have shown that blue-light-emitting diode (LED) LCUs have the potential to polymerise dental composites without having the drawbacks of halogen LCUs. Despite the relatively low irradiance of current LED LCUs, their e$ciency is close to that of conventional halogen LCUs with more than twice the irradiance. This phenomenon has not been explained fully yet. Hence, more tests of the LED LCU's e!ectiveness and of the mechanical properties of oral biomaterials processed with LED LCUs need to be carried out. This study investigates the #exural properties of three di!erent composites with three di!erent shades, which were polymerised with either a commercial halogen LCU or an LED LCU, respectively. In most cases no signi"cant di!erences in #exural strength and modulus between composites polymerised with a halogen LCU or an LED LCU, respectively, were found. A simple model for the curing e!ectiveness based on the convolution absorption spectrum of the camphorquinone photoinitiator present in composites and the emission spectra of the LCUs is presented.

Oxygenation measurements are widely used in patient care. However, most clinically available instruments currently consist of contact probes that only provide global monitoring of the patient (e.g., pulse oximetry probes) or local monitoring of small areas (e.g., spectroscopy-based probes). Visualization of oxygenation over large areas of tissue, without a priori knowledge of the location of defects, has the potential to improve patient management in many surgical and critical care applications. In this study, we present a clinically compatible multispectral spatial frequency domain imaging (SFDI) system optimized for surgical oxygenation imaging. This system was used to image tissue oxygenation over a large area (16×12 cm) and was validated during preclinical studies by comparing results obtained with an FDA-approved clinical oxygenation probe. Skin flap, bowel, and liver vascular occlusion experiments were performed on Yorkshire pigs and demonstrated that over the course of the experiment, relative changes in oxygen saturation measured using SFDI had an accuracy within 10% of those made using the FDA-approved device. Finally, the new SFDI system was translated to the clinic in a first-in-human pilot study that imaged skin flap oxygenation during reconstructive breast surgery. Overall, this study lays the foundation for clinical translation of endogenous contrast imaging using SFDI.

Transmission of compressed medical images is becoming a vital tool in telemedicine. Thus new methods are needed for efficient image compression. This study discovers the best design parameters for a data compression scheme applied to digital magnetic resonance (MR) images. The proposed technique aims at reducing the transmission cost while preserving the diagnostic information. By selecting the wavelet packet's filters, decomposition level, and subbands that are better adapted to the frequency characteristics of the image, one may achieve better image representation in the sense of lower entropy or minimal distortion. Experimental results show that the selection of the best parameters has a dramatic effect on the data compression rate of MR images. In all cases, decomposition at three or four levels with the Coiflet 5 wavelet (Coif 5) results in better compression performance than the other wavelets. Image resolution is found to have a remarkable effect on the compression rate.

Starting from the natural amino-acid L-tyrosine, a diphenolic monomeric molecule was developed using carbodiimide mediated solidphase synthesis techniques. This monomeric molecule was polymerized by reacting it in equimolar proportions with suitable dihalophosphates to yield novel biodegradable polyphosphates containing peptide linkages and phosphoester linkages alternating in the polymer backbone. The biodegradability of such a polymer is expected to arise from the hydrolytic degradability of the phosphoester linkages and the enzymatic degradability of the peptide linkages in the polymer backbone. Design of such a polymer is expected to make a significant contribution to biomaterials research, regarding drug delivery device and tissue engineering scaffold applications. The monomer was obtained by a novel solid phase carbodiimide-mediated amide coupling process. The subsequent polymers were obtained by solutionphase dehydrochlorination polycondensation reactions in the presence of a suitable acid acceptor. The synthesized polymers were characterized by 13 C NMR, 31 P NMR and FTIR for their chemical structure, by GPC for their molecular weight distribution, and by DSC and TGA for their thermal transition characteristics.

A theoretical study of possible neuromuscular incapacitation based on the application of high-intensity, ultrashort electric pulses is presented. The analysis is applied to a rat, but the approach is general and can be extended to any whole-animal and applies for any arbitrary pulse waveform. It is hypothesized that repeatable and reversible action potential blocks in nerves can be attained based on the electroporation mechanism. Our numerical studies are based on the Hodgkin-Huxley distributed circuit representation of nerves, and incorporate a nodal analysis for the time-dependent and volumetric perturbing potentials and internal electric fields in whole animals. The predictions are compared to actual 600-ns experimental reports on rats and shown to be in very good agreement. Effective strength-duration plots for neuromuscular incapacitation are also generated.

Optics has played a key role in the rapidly developing field of molecular imaging. The spectroscopic nature and high-resolution imaging capabilities of light provide a means for probing biological morphology and function at the cellular and molecular levels. While

This study was designed to evaluate the feasibility of using cylindrical ultrasound transducers mounted on a catheter for the ablation of cardiac tissues. In addition, the effects of ultrasound frequency and power was evaluated both using computer simulations and in vitro experiments. Frequencies of 4.5, 6, and 10 MHz were selected based on the simulation studies and manufacturing feasibility. These transducers were mounted on the tip of 7-French catheters and applied in vitro to fresh ventricular canine endocardium, submerged in flowing degassed saline at 37 degree C. When the power was regulated to maintain transducer interface temperature at 90-100 degree C, the 10-, 6-, and 4.5-MHz transducers generated a lesion depth of 5.9 +/- 0.2 mm, 4.6 +/- 1.0 mm, and 5.3 +/- 0.9 mm, respectively. The 10-MHz transducer was chosen for the in vivo tests since the maximum lesion depth was achieved with the lowest power. Two dogs were anesthetized and sonications were performed in both the left and right ventricles. The 10-MHz cylindrical transducers caused an average lesion depth of 6.4 +/- 2.5 mm. In conclusion, the results show that cylindrical ultrasound transducers can be used for cardiac tissue ablation and that they may be able to produce deeper tissue necrosis than other methods currently in use.

Decision making often occurs in the face of uncertainty about whether one's choices will lead to benefit or harm. The somatic-marker hypothesis is a neurobiological theory of how decisions are made in the face of uncertain outcome. This theory holds that such decisions are aided by emotions, in the form of bodily states, that are elicited during the deliberation of future consequences and that mark different options for behavior as being advantageous or disadvantageous. This process involves an interplay between neural systems that elicit emotional/ bodily states and neural systems that map these emotional/ bodily states.

Electrical bio-impedance can be used to assess skin cancers and other cutaneous lesions. The aim of this study was to distinguish skin cancer from benign nevi using multifrequency impedance spectra. Electrical impedance spectra of about 100 skin cancers and 511 benign nevi were measured. Impedance of reference skin was measured ipsi-laterally to the lesions. The impedance relation between lesion and reference skin was used to distinguish the cancers from the nevi. It was found that it is possible to separate malignant melanoma from benign nevi with 75% specificity at 100% sensitivity, and to distinguish nonmelanoma skin cancer from benign nevi with 87% specificity at 100% sensitivity. The power of skin cancer detection using electrical impedance is as good as, or better than, conventional visual screening made by general practitioners.

A hydrogel was prepared from pullulan and evaluated as a novel biomaterial for vascular engineering. Using a crosslinking process with sodium trimetaphosphate in aqueous solution, homogeneous, transparent, and easyto-handle pullulan gels were obtained with water-content higher than 90%. A circular punch was used to cut 6-mm diameter and 2-mm thickness discs for cell culture. Environmental scanning electron microscopy analysis of hydrated gels revealed a smooth surface, on which rabbit vascular smooth muscle cells were successfully seeded. The absence of cytotoxicity was evidenced by a live/dead assay. Fluorescence-labeled cells were observed adhering and progressively spreading out on the surface of the material. Cellular proliferation was followed for up to 1 week using an MTT assay. In addition, a complete in vitro degradation of the gels was achieved in 3 h upon incubation in a pullulanase solution (44 U/mL). In conclusion, we have shown the feasibility of preparing a biocompatible pullulan-based hydrogel that could support vascular cell culture. Based on these promising results, future studies will focus on the seeding of vascular cells on tubular-shaped hydrogels and the in vivo implantation of these new biomaterials.

Radiographic technology has advanced considerably during the last years with the advent of reconstruction techniques allowing visualization of slices through the body. In spite of the advantage of computed tomography compared to conventional radiographic methods, there are still some shortcomings with the method If a different section of the body is desired, another recording has to be made, the width of the dice reconstructed is fixed, and a full 1800 view angle is required.

Our aim is to apply image analysis on photosensitizer fluorescence and compare the relationship between histopathology and endoscopic fluorescence imaging. The correlation between hypericin fluorescence and histopathology of diseased tissue was explored in a clinical study involving 58 fluorescence cystoscopic images from 23 patients. Based on quantification of fluorescence colorimetric parameters extracted from the image analysis, diagnostic functions were developed to pathologically classify the bladder cancer. Our preliminary results show that the differences in fluorescence intensity ratios among the three different grades of bladder cancer are statistically significant. The results also show a decrease in macroscopic fluorescence intensity that correlated with higher cancer grades. By combining both the red-to-green and red-to-blue fluorescence intensity ratios into a 2-D scatter plot and defining diagnostic linear discrimination functions on the data points, this technique is able to yield an average sensitivity and specificity of around 68.6% and 86.1%, respectively, for pathological cancer grading of the three different grades of bladder cancer in our study. We conclude that our proposed approach in applying colorimetric intensity ratio analysis on hypericin fluorescence shows potential to optically grade bladder cancer in situ.

We describe a protocol to rapidly and reliably visualize blood vessels in experimental animals. Blood vessels are directly labeled by cardiac perfusion using a specially formulated aqueous solution containing 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI), a lipophilic carbocyanine dye, which incorporates into endothelial cell membranes upon contact. By lateral diffusion, DiI also stains membrane structures, including angiogenic sprouts and pseudopodial processes that are not in direct contact. Tissues can be immediately examined by conventional and confocal fluorescence microscopy. High-quality serial optical sections using confocal microscopy are obtainable from thick tissue sections, especially at low magnification, for three-dimensional reconstruction. It takes less than 1 h to stain the vasculature in a whole animal. Compared with alternative techniques to visualize blood vessels, including space-occupying materials such as India ink or fluorescent dye-conjugated dextran, the corrosion casting technique, endothelial cell-specific markers and lectins, the present method simplifies the visualization of blood vessels and data analysis.

In spite of the present advances in the scaffolds fabrication and bioreactor systems, the ability to create functional thick tissue masses in vitro is still a great tissue engineering challenge. To overcome this problem, the fabrication of a capillary bed, for nutrient supply to and waste product removal from the tissue engineering construct as it grows, is essential. However, the technical construction of a capillary-like architecture is complex and challenging. This study reports, for the very first time, a simple method to design and fabricate a porous hollow membrane sheet (PHMsh) to provide both a capillary bed and a scaffold to support tissue growth. The PHMsh composed of a flexible porous sheet involving parallel porous channels and can be used as flat-, rolled-, or sandwiched-shape scaffold. The PHMsh was fabricated from poly(ϵ-caprolactone) polymer solution using solvent casting methods (i.e., immersion precipitation and air casting). Optical microscopy and scanning electron microscopy were used for the morphological analyses. The PHMsh was surface treated using n-hepthylamine plasma polymer (HApp) and X-ray photoelectron spectroscopy confirmed successful surface coating. Human umbilical vein endothelial cells (HUVECs) and fibroblast cells were used to evaluate the capability of PHMsh toward cell adhesion. The HApp coating enhanced both HUVEC and fibroblast cells adhesion. The obtained preliminary results demonstrated the successful fabrication of the PHMsh, with potential application for tissue engineering scaffolds, particularly in large tissue mass generation under perfusion systems in vitro, which is our future research direction. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res, 2010

It remains unclear how metal released from implants affects cells of the immune system and, in particular, cells of the adaptive immune system, that is, T-helper lymphocytes. In this study, we investigated the effects of aluminum, chromium, cobalt, copper, iron, molybdenum, nickel, niobium, vanadium, and zirconium ions at concentrations from 0.05 to 5.0 mM on human CD4+ T lymphocytes. The DNA damage, apoptosis, necrosis, and proliferation responses of a human T-helper lymphocyte (Jurkat) cell line were evaluated to test our hypothesis that some metals will preferentially induce genotoxicity (DNA damage). Our results demonstrated that metal ions did not preferentially induce Jurkat T-lymphocyte DNA damage prior to other forms of toxicity, that is, apoptosis and/or direct necrosis. Nickel and vanadium induced the most DNA damage and were the most apoptotic metals tested, inducing >50% caspase-9 positive T cells at 0.05 mM and 0.1 mM concentrations, respectively. Cobalt and niobium were the most toxic metals, inducing <50% viability at ∼0.5 mM concentrations. Nickel and vanadium were the only metals to induce DNA damage at nearly the same concentrations that induced >50% apoptosis (i.e., <0.05 mM). All the metals tested induced T-cell apoptosis at a lower dose than that required to affect DNA damage or toxicity, implying that soluble metals released from implants may not be preferentially genotoxic to lymphocytes. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2008

This article provides a theoretical and methodological framework for matching, and ultimately the comprehensibility and usability of CPGs. Cognitive methods of analysis can help guideline designers and system the use of cognitive analysis to support the representation of biomedical builders throughout the development process, from the conceptual knowledge and the design of clinical systems, using clinical-practice design of a computer-based system to its implementation phases. By guidelines (CPGs) as an example. We propose that propositional and studying how guideline creators and developers represent guidelines, semantic analyses, when used as part of the system-development proboth mentally and in text, and how end-users understand and make cess, can improve the validity, usability, and comprehension of the decisions with such guidelines, we can inform the development of resulting biomedical applications. The framework we propose is based technologies that seek to improve the match between the representations on a large body of research on the study of how people mentally of experts and practitioners. We urge informaticians to recognize the represent information and subsequently use it for problem solving. potential relevance of cognitive analysis methods and to begin more This research encompasses many areas of psychology, but the more extensive experimentation with the their use in biomedical informatics important ones are the study of memory and the study of comprehenresearch. ᭧ 2001 Academic Press sion. Of particular relevance is research devoted to investigating the Key Words: cognitive analysis; propositional analysis; semantic netcomprehension and memory of language, expressed verbally or in text.

Temporomandibular disorders (TMD) are clinical manifestations of disorders causing pain and dysfunction in the Temporomandibular Joints (TMJs) and masticatory muscles. TMJ abnormalities cannot be reliably assessed by a clinical examination.Radiographic examination forms an integral component for the routine assessment in patients with temporomandibular joint disorders. Latest imaging modality is making life easy for those clinicians who must evaluate the hard and the soft tissue of the temporomandibular joint complex in patients with TMD's. Conventional radiographic TMJ projections like transpharyngeal, transcranial, panoramic radiograph, conventional tomographic sections of TMJ may be adequate in number of clinical situations but there are bony alterations that occur in these disorders like erosions, osteophytes, pneumatisation of articular eminence that are difficult to be detected in the conventional radiographs due to overlapping of anatomical structures. So the advanced imaging techniques such as MRI, Ultrasonography, CBCT are required for the understanding of diseases of Temporomandibular Joint. This paper aims to cover all the latest imaging modalities that can be helpful in the diagnosis of Temporomandibular Joint Disorders.

Based on permittivity changes, a new method to measure hematocrit (HCT) in extracorporeal blood systems is presented. Human blood samples were tested at different HCT levels pairing the values of permittivity change, obtained by means of a commercial impedance analyzer, with traditional centrifugation measurements. Data were correlated using both linear and nonlinear regression. When using the lineal model, the comparison

Measures of postural steadiness are used to characterize the dynamics of the postural control system associated with maintaining balance during quiet standing. The objective of this study was to evaluate the relative sensitivity of center-ofpressure (COP)-based measures to changes in postural steadiness related to age. A variety of time and frequency domain measures of postural steadiness were compared between a group of twenty healthy young adults (21-35 years) and a group of twenty healthy elderly adults (66-70 years) under both eyes-open and eyes-closed conditions. The measures that identified differences between the eyes-open and eyes-closed conditions in the young adult group were different than those that identified differences between the eye conditions in the elderly adult group. Mean velocity of the COP was the only measure that identified age-related changes in both eye conditions, and differences between eye conditions in both age groups. The results of this study will be useful to researchers and clinicians using COP-based measures to evaluate postural steadiness.

The capacity of bone for post-yield energy dissipation decreases with age. To gain information on the cause of such changes, we examined the mechanical behavior of human cadaveric bone as a function of progressive deformation. In this study, tensile specimens from tibiae of 9 middle aged and 8 elderly donors were loaded till failure in an incremental and cyclic (load-dwell-unload-dwellreload) scheme. The elastic modulus, maximum stress, permanent strain, stress relaxation, viscoelastic time constant, plastic strain energy, elastic release strain energy, and hysteresis energy were determined at incremental strains of each loading cycle. Experimental results showed that elderly bone failed at much lower strains compared to middle aged bone, but little age-related differences were observed in the mechanical behavior of bone until the premature failure of elderly bone. Energy dissipation and permanent strain appeared to linearly increase with increasing strain, while non-linear changes occurred in the modulus loss and stress relaxation/time constant with increasing strain. Such changes suggest that two distinct stages may exist in the progressive deformation of bone. In Stage I, rapid damage accumulation and increased involvement of collagen in load bearing appeared to dominate the mechanical behavior of bone with limited energy dissipation (<20% of total energy dissipated), whereas Stage II is dominated by continuous plastic deformation, accompanied by major energy dissipation through all three pathways till failure. This study suggests that damaging mechanisms in bone vary with deformation and age affects the post-yield mechanisms causing a significant decline in the capacity of aged bone to dissipate energy.

We have developed a new offline chromatographic approach for the selective enrichment of phosphorylated peptides that is directly compatible with subsequent analysis by online nano electrospray ionization tandem mass spectrometry. In this technique, a titanium dioxide (TiO 2 )-packed pipette tip is used as a phosphopeptide trap that acts as an offline first-dimension separation step in a twodimensional chromatography system. This is followed by online nano reversed-phase high-performance liquid chromatography. Here, we present suitable methods for enrichment, optimized separately for each step: sample loading, washing and elution from the TiO 2 -filled tips. To increase the trapping selectivity of the TiO 2 column, we used the sodium salt of 1-octanesulfonic acid combined with 2,5-dihydroxybenzoic acid as ion-pairing agents and displacers for acidic peptides. These agents also improve the binding of phosphorylated peptides and block the binding of non-phosphorylated ones. This enrichment procedure takes 30 min, followed by a 100-min HPLC program, including washing and an elution gradient.

A simple, high-throughput and specific high-performance liquid chromatography tandem mass spectrometry method has been developed and validated according to the FDA guidelines for simultaneous quantification of olmesartan and pioglitazone in rat plasma. The bioanalytical method consists of liquid–liquid extraction and quantitation by triple quadrupole mass spectrometry using electrospray ionization technique, operating in multiple reaction monitoring and positive ion modes. The compounds were eluted isocratically on a C18 column with a mobile phase consisting of a mixture of methanol and water (containing 0.5% formic acid) in a ratio of 9:1. The response to olmesartan and pioglitazone was linear over the range 0.01–10 µg/mL. The validation results demonstrated that the method had satisfactory precision and accuracy across the calibration range. Intra- and inter-day precisions ranged from 0.66 to 3.32 and from 0.94 to 2.93% (%CV), respectively. The accuracy determined at three quality control levels was within 91.27–107.28%. There was no evidence of instability of the analytes in rat plasma following the stability studies. The method proved highly reproducible and sensitive and was successfully applied in a pharmacokinetic study after single dose oral administration of olmesartan and pioglitazone to the rat. Copyright © 2010 John Wiley & Sons, Ltd.

Rigorous analysis of synaptic transmission in the central nervous system requires access to presynaptic terminals. However, cortical terminals have been largely inaccessible to presynaptic patch-clamp recording, due to their small size. Using improved patch-clamp techniques in brain slices, we recorded from mossy fiber terminals in the CA3 region of the hippocampus, which have a diameter of 2-5 lm. The major steps of improvement were the enhanced visibility provided by high-numerical aperture objectives and infrared illumination, the development of vibratomes with minimal vertical blade vibrations and the use of sucrose-based solutions for storage and cutting. Based on these improvements, we describe a protocol that allows us to routinely record from hippocampal mossy fiber boutons. Presynaptic recordings can be obtained in slices from both rats and mice. Presynaptic recordings can be also obtained in slices from transgenic mice in which terminals are labeled with enhanced green fluorescent protein.

A multi-channel wireless EEG (electroencephalogram) acquisition and recording system is developed in this work. The system includes an EEG sensing and transmission unit and a digital processing circuit. The former is composed of pre-amplifiers, filters, and gain amplifiers. The kernel of the later digital processing circuit is a micro-controller unit (MCU, TI-MSP430), which is utilized to convert the EEG signals into digital signals and fulfill the digital filtering. By means of Bluetooth communication module, the digitized signals are sent to the back-end such as PC or PDA. Thus, the patient&#39;s EEG signal can be observed and stored without any long cables such that the analogue distortion caused by long distance transmission can be reduced significantly. Furthermore, an integrated classification method, consisting of non-linear energy operator (NLEO), autoregressive (AR) model, and bisecting k-means algorithm, is also proposed to perform EEG off-line clustering at the back-end. ...

A common clinical practice during single-point Doppler analysis is to measure the centerline maximum velocity and to recover the time-averaged flow rate by exploiting an assumption on the shape of velocity profile (a priori formula), either a parabolic or a flat one. In a previous study, we proposed a new formula valid for the peak instant linking the maximum velocity and the flow rate by including a well-established dimensionless fluid-dynamics parameter (the Womersley number), in order to account for the hemodynamics conditions (Womersley number-based formula). Several in silico tests confirmed the reliability of the new formula. Nevertheless, an in vivo confirmation is missing limiting the clinical applicability of the formula. An experimental in vivo protocol using cine phase-contrast MRI (2-D PCMRI) technique has been designed and applied to ten healthy young volunteers in three different arterial districts: the abdominal aorta, the common carotid artery, and the brachial artery. Each PCMRI dataset has been used twice: 1) to compute the value of the blood flow rate used as a gold standard and 2) to estimate the flow rate by measuring directly the maximum velocity and the diameter (i.e., emulating the intravascular Doppler data acquisition) and by applying to these data the a priori and the Womersley number-based formulae. All the in vivo results have confirmed that the Womersley numberbased formula provides better estimates of the flow rate at the peak instant with respect to the a priori formula. More precisely, mean performances of the Womersley number-based formula are about three times better than the a priori results in the abdominal aorta, five times better in the common carotid artery, and two times better in the brachial artery.

Brachypodium distachyon is a novel model system for structural and functional genomics studies of temperate grasses because of its biological and genetic attributes. Recently, the genome sequence of the community standard line Bd21 has been released and the availability of an efficient transformation system is critical for the discovery and validation of the function of Brachypodium genes. Here, we provide an improved procedure for the facile and efficient Agrobacterium-mediated transformation of line Bd21. The protocol relies on the transformation of compact embryogenic calli derived from immature embryos using visual and chemical screening of transformed tissues and plants. The combination of green fluorescent protein expression and hygromycin resistance enables early identification of transformation events and drastically reduces the quantity of tissue to be handled throughout the selection process. Approximately eight independent fully developed transgenic Bd21 plants can be produced from each immature embryo, enabling the generation of thousands of T-DNA lines. The process-from wild-type seeds to transgenic T 1 seeds-takes B8 months to complete.

Using biodegradable scaffolds as an alternative to engineer new tissues has become an attractive candidate in various transplantation protocols. In particular, small intestinal submucosa (SIS), a dense connective matrix harvested from the small intestine, has gained attention due to a number of favorable properties. However, use of SIS is constrained by obtaining reliable, reproducible products in large-scale preparations that affect the regenerative process. To better understand the heterogeneous nature of SIS, this study focused on evaluating the location-dependent alterations in the physical characteristics of the matrices harvested from distal and proximal ends and processed inhouse (referred as hand-made). Additionally, results were compared with a commercially available machine-made Cook SIS. Tensile properties during monotonic loading and cyclical loading were compared in wet conditions. Further-more, permeability of these membranes to urea was analyzed using a custom-built chamber, and the microarchitecture was analyzed via scanning electron microscopy. These results showed that distal samples were more elastic and less permeable to urea relative to other samples. However, permeability in each sample was direction-dependent, that is, mucosal to serosal direction was less permeable compared to sorasal to mucosal direction in all the samples. Cook SIS was more susceptible to cyclical loading and had a shorter range of load carrying capacity. In summary, results show that physical characteristics of SIS are locationdependent.

The development of facile and rapid quantification of biologically active biomolecules such as isotretitoin in therapeutic drugs contained in many generic formulations is necessary for determining their efficiency and their quality to improve the human health care. Isotretritoin finds its applications in the maintenance of epithelial tissues. Different processes to date such as normal phase HPLC, or gas chromatrography among others are able to separate and quantify isotetroin. However, the extraction is quite complex and in the case of HPLC, the analysis requires long retention times. In such context, an isocratic reversedphase high-performance liquid chromatography (HP-LC) technique coupled with an UV-vis detector is described here for easy separation and quantification of 13-cis-retinoic (isotretinoin) from soft gelatin capsule formulations. The isotretinoin was extracted from three different commercial drug samples with tetrahydrofuran (THF) solvent by a procedure that can be completed in less than 10 minutes. Subsequent separation and quantification were accomplished in less than 5 minutes under isocratic reversed-phase conditions on a Lichrospher RP18 column and a mobile phase consisting of 0.01% TFA/acetonitrile (15/85, v/v) at a flow rate of 1.0 mL/min. Isotretoin was detected for the three samples via its UV-vis absorbance at 342 nm. The method was validated and the results showed good linearity, precision and accuracy for sensitive and selective quantitative determination of isotretinoin in the different pharmaceutical formulations. We found that the average isotretinoin content in two of the three commercial products fell outside the 90-110% United States Pharmacopeia specifications. Consequently, the facile extraction and the precise method for the biomole-cule quantification open up tremendous possibilities in improving the quality control of drugs which can exist as different generic brands.

Disturbances of the cerebrospinal fluid (CSF) flow in the brain can lead to hydrocephalus, a condition affecting thousands of people annually in the US. Considerable controversy exists about fluid and pressure dynamics, and about how the brain responds to changes in flow patterns and compression in hydrocephalus. This paper presents a new model based on the first principles of fluid mechanics. This model of fluid-structure interactions predicts flows and pressures throughout the brain's ventricular pathways consistent with both animal intracranial pressure (ICP) measurements and human CINE phase-contrast magnetic resonance imaging data. The computations provide approximations of the tissue deformations of the brain parenchyma. The model also quantifies the pulsatile CSF motion including flow reversal in the aqueduct as well as the changes in ICPs due to brain tissue compression. It does not require the existence of large transmural pressure differences as the force for ventricular expansion. Finally, the new model gives an explanation of communicating hydrocephalus and the phenomenon of asymmetric hydrocephalus.

As is known for centuries, humans exhibit an electrical profile. This profile is altered through various psychological and physiological processes, which can be measured through biosignals; e.g., electromyography (EMG) and electrodermal activity (EDA). These biosignals can reveal our emotions and, as such, can serve as an advanced man-machine interface (MMI) for empathic consumer products. However, such a MMI requires the correct classification of biosignals to emotion classes. This chapter starts with an introduction on biosignals for emotion detection. Next, a state-of-the-art review is presented on automatic emotion classification. Moreover, guidelines are presented for affective MMI. Subsequently, a research is presented that explores the use of EDA and three facial EMG signals to determine neutral, positive, negative, and mixed emotions, using recordings of 21 people. A range of techniques is tested, which resulted in a generic framework for automated emotion classification with up to 61.31% correct classification of the four emotion classes, without the need of personal profiles. Among various other directives for future research, the results emphasize the need for parallel processing of multiple biosignals.

We demonstrate the use of surface Zeta potential measurements as a new tool to investigate the interactions of iron oxide nanoparticles and cowpea mosaic virus (CPMV) nanoparticles with human normal breast epithelial cells (MCF10A) and cancer breast epithelial cells (MCF7) respectively. A substantial understanding in the interaction of nanoparticles with normal and cancer cells in vitro will enable the capabilities of improving diagnostic and treatment methods in cancer research, such as imaging and targeted drug delivery. A theoretical Zeta potential model is first established to show the effects of binding process and internalization process during the nanoparticle uptake by cells and the possible trends of Zeta potential change is predicted for different cell endocytosis capacities. The corresponding changes of total surface charge of cells in the form of Zeta potential measurements were then reported after incubated respectively with iron oxide nanoparticles and CPMV nanoparticles. As observed, after MCF7 and MCF10A cells were incubated respectively with two types of nanoparticles, the significant differences in their surface charge change indicate the potential role of Zeta potential as a valuable biological signature in studying the cellular Biomed Microdevices (interaction of nanoparticles, as well as specific cell functionality.

We propose and characterize a CMOS LSI-based neural stimulator for retinal prosthesis technology. The stimulator is based upon a multichip architecture in which small-sized CMOS stimulators named "unit chips" are organized on a flexible substrate. We designed a unit chip with an on-chip stimulator and light-sensing circuitry. We verified that all the functions implemented on the unit chip worked correctly and that an organized unit chip can be used as a retinal stimulator with multisite image-based patterned stimulation. We also demonstrated lightcontrolled retinal stimulation for the first time in an in vivo animal experiment on a rabbit's retina.

Several major costs associated with the production of biopharmaceuticals or vaccines in fermentation-based systems could be minimized by using plant chloroplasts as bioreactors, which facilitates rapid scale-up. Oral delivery of chloroplast-derived therapeutic proteins through plant cells eliminates expensive purification steps, low temperature storage, transportation and sterile injections for their delivery. Chloroplast transformation technology (CTT) has also been successfully used to engineer valuable agronomic traits and for the production of industrial enzymes and biomaterials. Here, we provide a detailed protocol for the construction of chloroplast expression and integration vectors, selection and regeneration of transformants, evaluation of transgene integration and inheritance, confirmation of transgene expression and extraction, and quantitation and purification of foreign proteins. Integration of appropriate transgenes into chloroplast genomes and the resulting high levels of functional protein expression can be achieved in B6 months in lettuce and tobacco. CTT is eco-friendly because transgenes are maternally inherited in most crop plants.