Development of a multispectral tissue characterization system for optimization of an implantable perfusion status monitor for transplanted liver.

Optimizing wavelength selection for monitoring perfusion during liver transplant requires an in-depth characterization of liver optical properties. With these, the impact of liver absorption and scattering properties can be investigated to select optimal wavelengths for perfusion monitoring. To accomplish this, we are developing a single integrating-sphere-based technique using a unique spatially resolved diffuse reflectance system for multispectral optical properties determination for thick samples. We report early results using a monochromatic source to measure the optical properties of well characterized tissue phantoms made from polystyrene spheres and Trypan blue. The presented results demonstrate the feasibility of using this unique system to measure optical properties of tissue phantoms. We are currently in the process of implementing an automated Levenberg-Marquardt diffuse-reflectance-profile fitting algorithm to enable near realtime robust computation of sample optical properties. Future work will focus on the incorporation of multispectral capability to provide needed data to facilitate development of more realistic liver tissue phantoms.

Pseudodielectric functions of uniaxial materials in certain symmetry directions.

The pseudodielectric function is often used to represent ellipsometric data and corresponds to the actual dielectric functions of materials when there is no surface overlayer and the material is isotropic. If a uniaxial material is oriented such that the optic axis is in the plane of incidence or is perpendicular to the plane of incidence, then the cross-polarization terms are zero and appropriate pseudodielectric functions can be determined from the ellipsometry data. We calculate the pseudodielectric functions for uniaxial crystals in three primary symmetry directions: (1) the optic axis is perpendicular to the plane of incidence, (2) the optic axis is in the plane of the sample surface and parallel to the plane of incidence, and (3) the optic axis is in the plane of the sample surface and perpendicular to the plane of incidence. These results are expanded in terms of the difference in the ordinary and extraordinary dielectric functions and compared with the approximation of Aspnes [J. Opt. Soc. Am. 70, 1275 (1980)]. Comparisons are made with experimental results on oriented crystals of rutile (TiO2), and a simple procedure is presented to determine the complex dielectric function from standard ellipsometry techniques.

Development of an implantable oximetry-based organ perfusion sensor.

A sensor system enabling real-time monitoring of organ perfusion following transplantation is presented. This system uses a three wavelength oximetry-based approach. The instrument is intended for implantation at the organ site during transplantation to provide real-time reporting of the perfusion status of the tissue for 7-10 days following the procedure. Data is transmitted from the sensor to a localized receiver using direct sequence spread spectrum techniques at 916 MHz. In this paper, the sensing method and associated electronics implementation are presented. The present status of system miniaturization is summarized along with plans for future miniaturization efforts. Preliminary sensor data is presented demonstrating the efficacy of the technique.

Enterobacterial repetitive intergenic consensus sequence-based PCR (ERIC-PCR); its ability to differentiate Streptococcus pyogenes strains and applicability to the study of outbreaks of streptococcal infection.

We evaluated the ability of enterobacterial repetitive intergenic consensus sequence-based PCR (ERIC-PCR) to differentiate 95 Streptococcus pyogenes strains with M or T serotypes isolated from sporadic streptococcal infections as compared with M or T serotypings and pulsed-field gel electrophoresis (PFGE). Although the ERIC-PCR had less discriminatory power, defined as the ability to divide the strains with the same serotypes into the different sub-types, than PFGE, it consistently classified the strains into 16 patterns with a high correlation with M or T serotyping. The PCR method further discriminated 4 M or T serotypes into sub-types. The application of ERIC-PCR to 5 outbreaks of streptococcal infection produced the results that agreed closely with those of T serotyping and PFGE. ERIC-PCR has sufficient discriminatory power and is a quick and relatively easy technique, making it useful for routine epidemiological investigations.

Measurement of the glucose transport time delay between the blood and aqueous humor of the eye for the eventual development of a noninvasive glucose sensor.

In the recent past, several noninvasive optically based methods have been proposed for physiologic glucose sensing. One proposed optical sensing site has been the eye, which, due to its unique optical properties, can be considered as a transparent optical window into the body. In particular, the aqueous humor within the anterior chamber of the eye has been shown to contain glucose levels correlated to those of blood. Concern, however, has been expressed that using the aqueous humor solution as a measure of blood glucose may be problematic due to the potential transport time delay between the blood and the aqueous humor glucose concentrations. This investigation was performed to measure the transport time delay in a rabbit model. The time delay between the blood and aqueous humor glucose concentrations was measured invasively in five New Zealand White rabbits over a series of weeks. An anesthesia protocol containing the drug Xylazine was used to elevate the blood glucose levels to a level commonly seen in diabetic patients. The difference between the glucose peak location times occurring in the blood and aqueous humor glucose response was measured and defined as the transport time delay. The average transport time lag was measured to be under 5 min. This measured time delay indicates that, indeed, the eye could potentially be used as a sensing site for indirect blood glucose measurements and may eventually aid the development of a noninvasive glucose sensor due to its unique optical properties compared to other biological tissues.

Comparison of noradrenergic and serotonergic antidepressants in reducing immobility time in the tail suspension test.

We examined the effects of two noradrenergic tricyclic antidepressants and two selective serotonin re-uptake inhibitors in the tail suspension test, with a suspension period of 30 min instead of the usual 10 min. Within the first 10 min, desipramine, nortriptyline and fluvoxamine significantly reduced the duration of immobility. Whereas desipramine and nortriptyline were also efficacious in the rest of the test period, fluvoxamine was not. Fluoxetine showed no significant effect throughout the study period. These results suggest that a prolonged tail suspension test results in functional changes in the noradrenergic and serotonergic systems and alters the sensitivity to antidepressants.

[Effects of fluvoxamine on both the desired anxiolytic effect and the adverse motor incoordination and amnesia induced by benzodiazepines].

Fluvoxamine, a selective serotonin reuptake inhibitor, is frequently used along with benzodiazepine anxiolytics in clinics. In this study, the effects of fluvoxamine on the anxiolytic effects as well as adverse effects of benzodiazepines were examined in the light/dark box, rota-rod and passive avoidance tests using mice. Diazepam, ethyl loflazepate and its active metabolite, CM7116, were used as benzodiazepine anxiolytics. The anxiolytic effects of diazepam, ethyl loflazepate and CM7116 were potentiated by intraperitoneal treatment with fluvoxamine at 10 mg/kg, whereas only those of ethyl loflazepate were potentiated by fluvoxamine at 45 mg/kg. The motor incoordination and amnesia induced by ethyl loflazepate and CM7116 were not affected by fluvoxamine, although these adverse effects of diazepam were potentiated by fluvoxamine at 45 mg/kg. Fluvoxamine itself showed no effects in any of the tests. These results suggest that low-dose fluvoxamine potentiates the anxiolytic effects of benzodiazepines, while high-dose fluvoxamine augments the adverse effects depending on the benzodiazepine used. Consequently, when fluvoxamine is administered along with benzodiazepines, the doses of both fluvoxamine and benzodiazepines should be carefully chosen to achieve anxiolytic effects without any adverse results.

Effects of rolipram, a selective inhibitor of phosphodiesterase 4, on hyperlocomotion induced by several abused drugs in mice.

The effects of rolipram, a selective inhibitor of phosphodiesterase 4, on the hyperlocomotion induced by several abused drugs (methamphetamine, morphine and phencyclidine) and a dopamine D1-receptor agonist (SKF81297; (+/-)-6-chloro-7,8-dihydroxy-1-phenyl-2,3 ,4,5-tetrahydro-1H-3-benzazepin hydrobromide) in mice were investigated. Methamphetamine (0.5-2.0 mg/kg), morphine (5.0-20 mg/kg), phencyclidine (1.25-5.0 mg/kg) and SKF81297 (2.5-10 mg/kg) each induced dose-dependent hyperlocomotion. A low dose (1.0 mg/kg) or moderate dose (3.2 mg/kg) of rolipram suppressed methamphetamine (2.0 mg/kg)- and morphine (20 mg/kg)-induced hyperlocomotion, but not phencyclidine (5.0 mg/kg)-induced hyperlocomotion. These results suggest that cAMP in the brain is involved in methamphetamine- and morphine-induced hyperlocomotion, while the underlying mechanism(s) of phencyclidine-induced hyperlocomotion may be different from those of methamphetamine- and morphine-induced hyperlocomotion. It is well known that methamphetamine- and morphine-induced hyperlocomotion are mediated by the dopaminergic system and that interaction between postsynaptic D1- and D2-receptors may play an important role in the expression of various dopamine-mediated behaviors. In the present study, SKF81297 (10 mg/kg)-induced hyperlocomotion was significantly but not completely suppressed by the highest dose of rolipram (10 mg/kg). Therefore it is unlikely that postsynaptic D1-receptor-mediated functions are involved in the suppressive effects of rolipram on methamphetamine- and morphine-induced hyperlocomotion. These results suggest that rolipram may inhibit methamphetamine- and morphine-induced hyperlocomotion via increase cAMP levels at D2-receptors.

Neuronal organization of the optic tectum in the river lamprey, Lampetra japonica: a Golgi study.

The neuronal and laminar organization of the optic tectum (OT) in the river lamprey was studied using the rapid Golgi method. Based primarily on the distribution pattern of the dendrites, the OT neurons were divided into vertical, horizontal and stellate neurons. The river lamprey OT shows a laminar structure consisting of eight concentric strata. The stratum ependymale consists of several rows of ependymal cells. The stratum cellulare periventriculare contains one to two rows of vertical neurons. The stratum fibrosum periventriculare is thin and contains a few vertical neurons. The stratum cellulare et fibrosum internum consists of several alternating cellular and fibrous layers: a large variety of vertical and horizontal neurons are distributed in this stratum. The stratum fibrosum centrale consists of compact horizontal fiber bundles, among which a few horizontal neurons are disseminated. In the stratum cellulare et fibrosum externum, numerous fibers run horizontally in a loosely organized plexus; various types of vertical, horizontal and stellate neurons are distributed among these fibers. The stratum opticum is the main terminal area of the optic nerve, and contains stellate and horizontal neurons. The stratum marginale is a thin layer and consists of sparse populations of vertical and horizontal neurons. Besides the above outer to inner laminar structure, the OT is divided into medial (m-OT) and lateral parts (1-OT), based primarily on the distribution pattern of the dendrites. The dendrites of neurons in the m-OT are distributed almost exclusively within the OT. On the other hand, the dendrites of some neurons in the 1-OT extended into the confines of the torus semicircularis (TS), and conversely, the dendrites of some neurons in the TS are distributed in the 1-OT. These findings are discussed in relation to the neuronal and laminar organization of the OT in other lamprey species and to recent hodological studies.

Inhibitory effects of rolipram on partially purified phosphodiesterase 4 from rat brains.

Several previous studies have demonstrated that the phosphodiesterase 4 selective inhibitor rolipram affects cellular function at a much lower concentration than the reported Ki value for phosphodiesterase 4 inhibition. In this study, we examined the inhibitory effect of rolipram on rat brain phosphodiesterase 4 to determine the heterogeneity of the enzyme activity. Partial purification of various phosphodiesterases from the rat brain was performed by anion-exchange chromatography. The eluant was pooled into four fractions, two of which manifested cAMP-selective phosphodiesterase activity that was blocked by 10 microM of rolipram, indicating the presence of phosphodiesterase 4 in these fractions. The IC50 of rolipram (racemate) of these two fractions was 492 and 79 nM, respectively. The R-(-)-enantiomer of rolipram inhibited the cAMP-phosphodiesterase activity in the latter fraction 10 times more than did S-(+)-rolipram, and the inhibition of the former fraction was less stereospecific. Dixon plot analysis revealed that the rolipram enantiomers inhibited the cAMP-phosphodiesterase in the latter fraction in a multiphasic manner, with two Ki values, one at the micromolar level and the other at the sub-micromolar level, respectively, for both of the enantiomers. These results suggest that there is a heterogeneity for phosphodiesterase 4 in the rat brain, and some of the phosphodiesterase forms are sensitive to rolipram.

Neuronal organization of the spinal cord in the red stingray (Dasyatis akajei: chondrichthyes).

The neuronal organization of the spinal cord in red stingray was studied using the rapid Golgi method. The gray matter of the spinal cord was divided into seven laminae: RS-I, RS-II, RS-III, RS-IV, RS-V, RS-VI and RS-VII. RS-I is cell dense lamina which occupies the major part of the dorsal horn and corresponds to laminae I and II of the spinal cord of mammals, birds and reptiles. The neurons of the lamina I are interspersed with those of lamina II, without forming a discrete lamina. RS-II is located at the base of the dorsal horn and is considered to correspond to the nucleus proprius. RS-III and IV form the intermediate zone and are highly reticulated. A few neurons of various shapes and sizes are distributed among the numerous fibers. The nuclei such as the intermediolateral, intermediomedial or Clarke's nucleus cannot be identified in the intermediate zone. RS-V and VI constitute the ventral horn. RS-V occupies the major part of the ventral horn and contains motoneurons which are distributed diffusely, without forming any distinct cell groups. RS-VI is located in the ventromedial part of the ventral horn, contains commissural neurons and correspond to lamina VIII. RS-VII is a small area surrounding the central canal and corresponds to lamina X. Thus, while the major features of the spinal cord of the red stingray can be correlated with those of the spinal cord of mammals, birds and reptiles, the neuronal organization of the spinal cord of the red stingray remains in an undifferentiated state.

Distribution of the parathyroid hormone-related peptide and its receptor in the saccus vasculosus and choroid plexus in the red stingray (Dasyatis akajei: Elasmobranch).

1. The exact role of the parathyroid hormone-related peptide (PTHrP) is not fully understood. We used immunohistochemistry to localize the PTHrP and its receptor in the brain of the red stingray, particularly in the saccus vasculosus (SV) and choroid plexus. 2. Immunoreactive PTHrP and its receptor were detected in the epithelial cells of the SV and the choroid plexus. In addition, the neuronal perikarya in the nucleus of the SV located in the hypothalamus is positive for the PTHrP. 3. No PTHrP-containing neurons were detected in the choroid plexus. Extracts of SV and choroid plexus showed positive reactions against the PTHrP and its receptor antibody in Western blot analysis. 4. High levels of immunoreactive PTHrP were detected in the plasma equivalent to those present in human humoral malignant hypercalcemia. In contrast, the immunoreactive PTHrP concentration in the cerebrospinal fluid was below detectable levels. 5. Our results suggest that the regulation of the PTHrP in the SV differs from that in the choroid plexus in the red stingray.

Neuronal organization of the olfactory bulb in the hagfish, Eptatretus burgeri: a Golgi study.

The neuronal organization of the olfactory bulb (OB) in the hagfish (Eptatretus burgeri) was studied using the rapid Golgi method. Cytologically, two groups of cells, the mitral and stellate cells, were discernible in the OB. Cytoarchitecturally, the OB showed a distinct laminar structure. From the periphery inward, the following four strata were distinguished: the stratum nervosum, stratum glomerulosum, stratum mitrale and stratum stellatum. Olfactory fibers from the olfactory epithelium reach the rostral aspect of the OB and form the stratum nervosum. The olfactory fibers run deeply in the OB, enter the stratum glomerulosum and terminate in the olfactory glomeruli which are arranged in three to four rows. Numerous mitral and a few stellate cells are distributed in periglomerular areas. The dendrites of the mitral cells terminate in one to two glomeruli in tufted terminals, while those of the stellate cells are distributed in periglomerular areas. The stratum mitrale also consists of mitral and stellate cells. The mitral cells in this stratum extend long dendrites to 4-5 widely separated glomeruli and generate axons traveling caudally. The dendrites of the stellate cells are long and are distributed in the stratum glomerulosum and stratum stellatum, as well as within the stratum mitrale. The stratum stellatum occupies a narrow caudal area and consists mainly of stellate cells extending long dendrites to the stratum stellatum and stratum mitrale.

Phosphodiesterase type 4 that regulates cAMP level in cortical neurons shows high sensitivity to rolipram.

To characterize the role of phosphodiesterase type 4 (a cAMP-specific and rolipram-sensitive phosphodiesterase) among phosphodiesterases in the regulation of the intracellular cAMP level in cortical neurons, we investigated the effects of phosphodiesterase inhibitors on the intracellular cAMP levels in primary cultured rat cortical neurons. Selective inhibitors of phosphodiesterase type 4 and type 2 significantly enhanced beta-adrenoceptor-mediated cAMP increase. Selective inhibitors of phosphodiesterase type 1, type 3 and type 5/6 had no effect on the cAMP level. Rolipram enhanced the beta-adrenoceptor-mediated cAMP increase in cortical neurons, astrocytes and vascular smooth muscle cells at different minimum effective concentrations (10, 100 and 1000 nM, respectively). These findings indicate that phosphodiesterase type 4, showing a high-sensitivity to rolipram, plays a major role in regulating cAMP in the cortical neurons, and that rolipram at low doses enhances the intracellular cAMP increase in the cortical neurons selectively.

Rolipram, a phosphodiesterase-4-selective inhibitor, promotes the survival of cultured rat dopaminergic neurons.

We evaluated the effects of rolipram, a selective inhibitor of phosphodiesterase (PDE) 4, on the survival of dopaminergic neurons in 13-day culture. Rolipram did not affect the survival of dopaminergic neurons in the absence of forskolin, but significantly enhanced the survival of dopaminergic neurons in the presence of 10(-5) M forskolin in a concentration-dependent manner (10(-8) - 10(-5) M). Rolipram also enhanced the neurotrophic effect of forskolin on total neurons including dopaminergic and non-dopaminergic neurons at a high concentration (10(-5) M), but did not affect the survival of cells containing glutamate or gamma-aminobutylic acid. A non-selective PDE inhibitor, 1-isobutyl-3-methylxanthine, caused a marked increase of dopaminergic neurons, whereas selective inhibitors of PDE2 and PDE3 showed far weaker effects. A PDE1 inhibitor, on the other hand, caused non-specific cell death in the presence or absence of forskolin. These findings suggest that rolipram has a potential to enhance the survival of dopaminergic neurons selectively by way of PDE4 inhibition.

Rolipram, a selective inhibitor of phosphodiesterase type 4, pronouncedly enhanced the forskolin-induced promotion of dopamine biosynthesis in primary cultured rat mesencephalic neurons.

A selective inhibitor of cyclic nucleotide phosphodiesterase (PDE) 4, rolipram, markedly enhanced the forskolin-induced increase of intracellular dopamine and dihydroxyphenylacetate (DOPAC, a metabolite of dopamine) levels in primary cultured rat mesencephalic neurons and the forskolin-induced increase of dopamine and DOPAC in extracellular medium. Selective inhibitors of PDE2, PDE3, PDE5 and PDE6 did not have such a promoting effect, and the PDE1 inhibitor vinpocetine and W-7 caused dopamine depletion in the neurons. These findings suggested that PDE4 plays a major role in regulating the intracellular cAMP level to control the dopamine biosynthesis in mesencephalic neurons, whereas PDE1 regulates dopamine release instead.

Percutaneous transluminal coronary angioplasty for morphologic left anterior descending artery lesion in a patient with dextrocardia. A case report and literature review.

A case report of percutaneous transluminal coronary angioplasty (PTCA) to treat coronary atherosclerotic lesions in a patient with dextrocardia associated with situs inversus totalis is presented. The patient was a sixty-two-year-old man who was admitted with a diagnosis of congestive heart failure. Cardiac catheterization was performed. Left ventriculography showed mild hypokinesis in segments 2 and 3 with ejection fraction of 63%. Coronary arteriography revealed 74% stenosis in segment 7 of the left anterior descending (LAD) artery. PTCA for this lesion was performed. Successful dilation was achieved with the residual stenosis in the LAD reduced from 74% to 34%. Performance of PTCA in patients with dextrocardia is extremely rare, and only 8 cases have been reported to date. However, by visualizing the procedure as a mirror image and choosing a guide catheter that permits good engagement, it appears possible to perform it like ordinary PTCA.

Unusual right coronary artery runoff with right and left main coronary artery aneurysm. A case report.

A rare case of unusual right coronary artery runoff with right and left main coronary artery aneurysm is presented. The discussion is mainly from the etiologic point of view.

Comparison of agar-gel precipitin responses among strains of fowl adenovirus using antigens prepared from chorioallantoic membranes and chicken kidney cell cultures.

Agar-gel precipitin responses obtained for serologically different strains of fowl adenovirus (FAV) in tests using antigens prepared from FAV-infected chorioallantoic membranes (CAM antigen) and chicken kidney cell cultures (CKC antigen) were compared. Findings showed that both types of antigens exhibited less sensitivity to heterologous than to homologous antisera and that quantitative differences in sensitivity were present between serotypes. CAM antigens were more sensitive than CKC antigens to heterologous antisera. Polyvalent CAM antigens containing 2 or 3 antigens increased sensitivity in testing of field serum samples, resulting in a higher rate of detection.