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Crystal structure of the Atx1 metallochaperone protein at 1.02 A resolution.

BACKGROUND: Metallochaperone proteins function in the trafficking and delivery of essential, yet potentially toxic, metal ions to distinct locations and particular proteins in eukaryotic cells. The Atx1 protein shuttles copper to the transport ATPase Ccc2 in yeast cells. Molecular mechanisms for copper delivery by Atx1 and similar human chaperones have been proposed, but detailed structural characterization is necessary to elucidate how Atx1 binds metal ions and how it might interact with Ccc2 to facilitate metal ion transfer. RESULTS: The 1.02 A resolution X-ray structure of the Hg(II) form of Atx1 (HgAtx1) reveals the overall secondary structure, the location of the metal-binding site, the detailed coordination geometry for Hg(II), and specific amino acid residues that may be important in interactions with Ccc2. Metal ion transfer experiments establish that HgAtx1 is a functional model for the Cu(I) form of Atx1 (CuAtx1). The metal-binding loop is flexible, changing conformation to form a disulfide bond in the oxidized apo form, the structure of which has been solved to 1.20 A resolution. CONCLUSIONS: The Atx1 structure represents the first structure of a metallochaperone protein, and is one of the largest unknown structures solved by direct methods. The structural features of the metal-binding site support the proposed Atx1 mechanism in which facile metal ion transfer occurs between metal-binding sites of the diffusible copper-donor and membrane-tethered copper-acceptor proteins. The Atx1 structural motif represents a prototypical metal ion trafficking unit that is likely to be employed in a variety of organisms for different metal ions.

Acid Anhydride Hydrolases↗

Prolonged anti-inflammatory action of DL-lactide/glycolide copolymer nanospheres containing betamethasone sodium phosphate for an intra-articular delivery system in antigen-induced arthritic rabbit.

PURPOSE: The objective of the present study was to develop prolonged anti-inflammatory action of DL-lactide/glycolide copolymer (PLGA) nanosphere incorporating a water-soluble corticosteroid (betamethasone sodium phosphate; BSP). Another aim was to demonstrate the biocompatibility and biologic efficacy of these BSP-loaded nanospheres directly administered into ovalbumin-induced chronic synovitis in the rabbit. METHODS: BSP-loaded nanospheres were prepared by an emulsion solvent diffusion method in oil (caprylate and caprate triglyceride). The drug releasing properties of the nanospheres were measured in vitro in phosphate buffer saline (PBS: pH7.4), and in vivo in rat air-pouch (pseudo synovial fluid). The BSP-loaded nanosphere suspensions were administered into the joint cavity in a model of antigen-induced arthritic rabbit and evaluated by measuring the joint swelling, and the biocompatibility was appraised by histologic microscopy. RESULTS: The BSP-nanospheres were a unimodally-dispersed particulate system with a mean diameter ranging from 300 to 490 nm, and BSP was efficiently entrapped in the lipophilic copolymer (PLGA), although its hydrophilic properties. The drug release-rate from the nanospheres in PBS was controlled by the molecular weight and the lactic/glycolic acid (LA/GA) ratio of the polymers. The in vitro releasing study demonstrated that sustained drug release occurred for over three weeks. In the antigen-induced arthritic rabbit, the joint swelling decreased significantly by administering BSP-loaded nanospheres during a 21-day period after intra-articular challenge. With regards to the prolonged anti-inflammatory efficacy, serum antibody to ovalbumin showed a sustained reduction during the period, and the steroidal effect appeared by the degradation of the polymer in the synovium. The BSP-nanosphere administered was phagocytosed by the synovial activated-cells and the cartilage degradation was almost prevented. CONCLUSIONS: Direct intra-articular injection of a PLGA nanosphere system with a water-soluble steroid provided a prolonged pharmacological efficacy in the joints of arthritic rabbits. The local anesthetic in the knee-joints was evaluated to be safe and without biologic damage.

Animals↗

In vivo noninvasive determination of abnormal water diffusion in the rat brain studied in an animal model for multiple sclerosis by diffusion-weighted NMR imaging.

In vivo NMR images of the rat brain were obtained using a NMR microscope (7 T) from SMIS (England). Four animals were imaged every 3-4 days during a pathological cycle (starting after induction and up to 37 days) of experimental allergic encephalomyelitis (EAE), an animal model for multiple sclerosis. The EAE rats were weighted and clinically scored daily. We aimed at measuring the apparent diffusion coefficient (ADC) or the mean diffusivity (D) with a high accuracy, and within a reasonable experimental time frame, because of the clinical situation of the animals. Therefore, we fitted the ADC value from five diffusion-weighted images--with an experimental time of 17 min/image--and chose to apply diffusion-sensitizing gradients in a direction intersecting all fiber directions of the external capsule. With this, we also obtained high b-values. For the control rats, we obtained a statistical mean value of ADC = (388 +/- 16) 10(-12) m2/s for gray matter and a statistical mean value of (D) of (750 +/- 30) 10(-12) m2/s for white matter, measured in the external capsule. For the EAE rats, no alterations in ADC values of gray matter with increasing clinical scores were observed. Concerning white matter, as determined in the external capsule, there were no significant differences in (D) values between controls and EAE rats before clinical signs occurred. However, when clinical signs were observed, we could demonstrate a significant positive correlation between the clinical score and the (D) values in the external capsule. As the clinical signs became more severe, we measured a rise in water diffusion (increase in (D)) in the external capsule, which was accompanied by the occurrence of interstitial edema as revealed by a complementary histological study.

Animals↗

MRI-based finite-element analysis of left ventricular aneurysm.

Tagged MRI and finite-element (FE) analysis are valuable tools in analyzing cardiac mechanics. To determine systolic material parameters in three-dimensional stress-strain relationships, we used tagged MRI to validate FE models of left ventricular (LV) aneurysm. Five sheep underwent anteroapical myocardial infarction (25% of LV mass) and 22 wk later underwent tagged MRI. Asymmetric FE models of the LV were formed to in vivo geometry from MRI and included aneurysm material properties measured with biaxial stretching, LV pressure measurements, and myofiber helix angles measured with diffusion tensor MRI. Systolic material parameters were determined that enabled FE models to reproduce midwall, systolic myocardial strains from tagged MRI (630 +/- 187 strain comparisons/animal). When contractile stress equal to 40% of the myofiber stress was added transverse to the muscle fiber, myocardial strain agreement improved by 27% between FE model predictions and experimental measurements (RMS error decreased from 0.074 +/- 0.016 to 0.054 +/- 0.011, P < 0.05). In infarct border zone (BZ), end-systolic midwall stress was elevated in both fiber (24.2 +/- 2.7 to 29.9 +/- 2.4 kPa, P < 0.01) and cross-fiber (5.5 +/- 0.7 to 11.7 +/- 1.3 kPa, P = 0.02) directions relative to noninfarct regions. Contrary to previous hypotheses but consistent with biaxial stretching experiments, active cross-fiber stress development is an integral part of LV systole; FE analysis with only uniaxial contracting stress is insufficient. Stress calculations from these validated models show 24% increase in fiber stress and 115% increase in cross-fiber stress at the BZ relative to remote regions, which may contribute to LV remodeling.

Animals↗

The chemotactic response of tumor cells. A model for cancer metastasis.

Injection of a C5-derived chemotactic factor for tumor cells into the peritoneal cavities of Sprague-Dawley rats induced diffuse mesenteric metastasis following the intravenous injection of Walker carcinosarcoma cells. Intraperitoneal injections of culture medium, histamine, or of trypsin-treated albumin resulted in many fewer metastases. Intraperitoneal injections of the chemotactic factor, unlike histamine, did not alter mesenteric vasopermeability as measured by the exudation of Evans blue into the mesentery. In vitro, tumor cells responded to the chemotactic factor by demonstrating directed migration in the Boyden chamber, by volume changes, measurable in the Coulter counter, and by demonstrating an increased adherence to nylon fibers. These phenomena are similar to the behavior of neutrophils in the presence of their chemotactic factors. All the responses in vitro were markedly depressed by the addition of 2-deoxyglucose, while the cell swelling response was slightly enhanced by cytochalasin B (again similar to the responses of leukocytes). The data suggest that movement of tumor cells from the circulation may be under chemotactic influence in the manner similar to the responsiveness of neutrophils to leukotactic stimuli in vivo.

Animals↗

Leaf display and photosynthesis of tree seedlings in a cool-temperate deciduous broadleaf forest understorey.

To examine a possible convergence in leaf photosynthetic characteristics and leaf display responses to light environment in seedlings of three canopy and two shrub tree species in understorey of cool-temperate deciduous broadleaf forest, relationships between light environment, leaf orientation and leaf light-photosynthetic response were measured. Light capture of the seedlings (17-24 individuals with 2-12 leaves for each species) was assessed with a three dimensional geometric modeling program Y-plant. Leaf photosynthetic characteristics of the five species were found to have acclimated to the understorey light environment, i.e., low light compensation point and high apparent quantum yield. In addition, light-saturated photosynthetic rates were higher in seedlings inhabiting microsites with higher light availability. Efficiencies of light capture and carbon gain of the leaf display were evaluated by simulating the directionalities of light capture and daily photosynthesis for each seedling using hemispherical canopy photography. The results showed that most of the seedlings orientated their leaves in a way to increase the daily photosynthesis during the direct light periods (sunflecks) rather than maximize daily photosynthesis by diffuse light. Simulations also showed that daily photosynthesis would increase only 10% of that on actual leaf display when the leaves orientated to maximize the diffuse light interception. Simulations in which leaf orientations were varied showed that when the leaf display fully maximized direct light interception, the time that leaves were exposed to excessive photon flux density of >800 mumol photons m(-2) s(-1) were doubled. The understorey seedlings studied responded to the given light environments in a way to maximize the efficiency of acquisition and use of light during their short (approximately 3 month) seasonal growth period.

Computer Simulation↗

The state of head injury biomechanics: past, present, and future: part 1.

This article is the first of two parts of a comprehensive survey of the biomechanics of head injury since its inception in 1939 in the United States, the separation being made for temporal and spatial reasons. The second portion of this material will be published at a later time in this journal. The discussion will be almost exclusively limited to nonpenetrating events. The topics presented in the following sections include an introduction that discusses the magnitude of the problem, the basic tools of biomechanics, and significant major reference sources covering this subject. This is succeeded by a brief description of the components of the head, classification of head injuries, early experimental investigations and human tolerance considerations, measurement techniques of kinetic parameters, and head motion and head injury investigations prior to 1966. A Head Injury Conference sponsored by the National Institutes of Neurological Diseases and Stroke in 1966 changed the landscape of investigations in this area. While informal collaboration between neurosurgeons and engineers had existed prior to this time, the conference established a permanent mechanism of synergism between these disciplines, produced the first zero-order realistic model of biomechanical head injury investigation, and established a 4-year program of federally funded research into the mechanical properties of the tissues of the cranium. While a recession precluded a continuation of the national sponsorship of such work, this 4-year period of intensive research resulted in a nationwide individual effort to develop further knowledge in this area. The current presentation, then, covers the mechanical and structural properties of solid and fluid tissues of the head, emphasizing progress during the past 3 decades; fetal cranial properties; analytical and numerical head injury models; experimental cranial loads applied to human volunteers and cadaver heads, dynamic loading of surrogate heads; and, finally, head injury mechanisms. The future publication will encompass experimental, analytical, and some numerical and regulatory information and that will be divided into the following sections: 1. head injury experimentation involving translatory and rotational motion: equipment, subjects and mechanical and physiological consequences 2. diffuse axonal injury: production and traumatic effects; mechanical properties at the axonal and neuronal level 3. vehicular crash investigation and simulation: reconstruction methodologies, staging, surrogate validation, and occupant protection, including vehicular design 4. injury thresholds and tolerances, including skull and vessel failure and brain and brainstem damage, including consideration of loading directions 5. criteria for head injury: governmental and industry regulations, including effects of combined motion- and tissue-level loading 6. further discussions of cranial component properties and injury mechanisms 7. sports head injury considerations: boxing, baseball, softball, football, ice hockey, and skiing activities; protective head devices for these activities 8. vehicular protective devices: design, efficacy, standards, and limitations; models for helmets and experimental validation. This presentation is based on my nearly 4 decades of head injury research, continuous collaboration and discussions with prominent members of the neurosurgical and orthopedic community, and an exhaustive, 2-year search of the literature. While every effort has been made to include all relevant information, it is inevitable that some important research has not come to my attention, and I apologize for any such omissions. It is hoped that this survey will serve as a resource for researchers and practitioners in the area of traumatic head injury and provide a roadmap for further investigations that are urgently needed. For example, this could include a determination of the rate of absorption of blood emitted from broken vessels, and, hopefully, some correlation between mechanical failure and physiological dysfunction of the various relevant tissues of the head. Although a good beginning has been initiated, additional information at the neuronal and axonal level concerning the effect of loading on function as well as age-related changes in geometry and tissue properties is also needed.

Biomechanical Phenomena↗

[Changes of extracellular potassium concentration in the cortex and brain stem during the acute phase of experimental closed head injury (author's transl)].

The high concentration of extracellular potassium ((K+)o) impedes neuronal activity by depolarizing the membrane potential and further causing depolarization block or conduction block, and also causes swelling of astrocytes, which may result in narrowing of extracellular space and affect the diffusion of metabolites. Thus, when high concentration of extracellular potassium prolongs, pathological changes of neurons may ensue. In such state as cerebral ischemia, contusion or spreading depression, marked elevation of extracellular potassium was reported by many authors. In order to reveal the changes of extracellular potassium during the acute phase of closed head injury, following experimental studies were performed. In the first experiment, closed head injury model of the mouse was used. Directly after the impact with 600 g.cm, spontaneous movement of the animal disappeared for about 5-6 minutes often accompanied by immediate epilepsy. In this state, cortical (K+)0 increased from the control level of 4.1 +/- 1.8 mM to 20-30 mM. The elevated (K+)0 recovered gradually within 10-30 minutes. Directly after the same impact, apnea, bradycardia and low voltage EEG also appeared. In the second experiment, closed head injury model of the rat was used. With the impact of 9000 g.cm, the same concussion-like symptoms appeared as those of mice. Directly after the impact, (K+)0 in the cortex and brain stem elevated but the change patterns of cortical (K+)0 and brain stem (K+)0 were different. The different patterns in changes of (K+)0 were classified into four types. Type 1. Both cortical and brain stem (K+)0 elevated significantly immediately after the trauma, and recovered to control level within 30 minutes. Type 2. Both (K+)0 elevated significantly as in type 1 but only brain stem (K+)0 recovered within 30 minutes. Type 3. With the exacerbation of vital signs, both (K+)0 elevated over 50 mM, resulting in the death of the animal. Type 4. Although cortical (K+)0 elevated significantly after the impact, the increase of brain stem (K+)0 was below 10 mM. In type 1,2 and 3, blood pressure always elevated and brady- or apnea appeared immediately after the impact. In type 4, blood pressure after the impact showed to become slightly lower transiently as compared to control blood pressure. In control study, spreading depression was elicited by application of KC1-solution on the cortex. Spreading depression caused the elevation of cortical (K+)0 only, and almost no changes of brain stem (K+)0 were observed. The present study suggested that during the acute phase of closed head injury, diffuse neuronal disturbances involving the brain stem existed. And the elevation of brain stem (K+)0 is an important finding of this study because it may affect the level of consciousness directly by the mechanisms as described above.

Animals↗

IL-6 transgenic mouse model for extraosseous plasmacytoma.

Plasma cell neoplasms in humans comprise plasma cell myeloma, otherwise known as multiple myeloma, Ig deposition and heavy chain diseases, and plasmacytoma (PCT). A subset of PCT, designated extramedullary PCT, is distinguished from multiple myeloma and solitary PCT of bone by its distribution among various tissue sites but not the bone marrow. Extramedullary (extraosseus) PCT are rare spontaneous neoplasms of mice but are readily induced in a susceptible strain, BALB/c, by treatment with pristane. The tumors develop in peritoneal granulomas and are characterized by Myc-activating T(12;15) chromosomal translocations and, most frequently, by secretion of IgA. A uniting feature of human and mouse plasma cell neoplasms is the critical role played by IL-6, a B cell growth, differentiation, and survival factor. To directly test the contribution of IL-6 to PCT development, we generated BALB/c mice carrying a widely expressed IL-6 transgene. All mice exhibited lymphoproliferation and plasmacytosis. By 18 months of age, over half developed readily transplantable PCT in lymph nodes, Peyer's patches, and sometimes spleen. These neoplasms also had T(12;15) translocations, but remarkably, none expressed IgA. Unexpectedly, approximately 30% of the mice developed follicular and diffuse large cell B cell lymphomas that often coexisted with PCT. These findings provide a unique model of extramedullary PCT for studies on pathogenesis and treatment and suggest a previously unappreciated role for IL-6 in the genesis of germinal center-derived lymphomas.

Animals↗

H fluxes in excised samanea motor tissue : I. Promotion by light.

Previous investigators revealed that white light-promoted leaflet opening in Samanea saman (Jacq) Merrill depends upon K(+) uptake by extensor cells and efflux from flexor cells of the pulvinus, while dark-promoted closure depends upon K(+) fluxes in the opposite directions. We now monitored H(+) fluxes during pulvinar movement to test a model proposing coupled H(+)/K(+) fluxes. H(+) fluxes were monitored by measuring changes in the pH of a weakly buffered solution (initial pH = 5.5) bathing excised strips of extensor or flexor tissue. White light at hour 3 of the usual dark period promoted pulvinar opening, H(+) efflux from extensor cells and uptake by flexor cells, while darkness at hours 2 to 4 of the usual light period promoted pulvinar closure, H(+) uptake by extensor cells and efflux from flexor cells. The following conditions altered H(+) fluxes during dark-promoted closure. (a) Light reversed the directions of the fluxes in both extensor and flexor cells. (b) Anoxia increased the rate of H(+) uptake by extensor cells and promoted H(+) uptake (rather than efflux) by flexor cells, consistent with an outwardly directed H(+) pump. KCN showed similar effects initially, but they were transient. (c) Increase in external pH from 5.5 to 6.7 promoted H(+) efflux (rather than uptake) by extensor cells and increased the rate of H(+) efflux from flexor cells, presumably by decreasing the rate of inward diffusion. (d) Change in external K(+) did not alter H(+) fluxes by extensor cells, but removal of external K(+) decreased the rate of H(+) efflux from flexor cells by 70%. These observations support a model for coupled H(+)/K(+) fluxes in pulvinar cells during light-and dark-promoted leaflet movements.

Journal Article↗

Orthogonal tensor invariants and the analysis of diffusion tensor magnetic resonance images.

This paper outlines the mathematical development and application of two analytically orthogonal tensor invariants sets. Diffusion tensors can be mathematically decomposed into shape and orientation information, determined by the eigenvalues and eigenvectors, respectively. The developments herein orthogonally decompose the tensor shape using a set of three orthogonal invariants that characterize the magnitude of isotropy, the magnitude of anisotropy, and the mode of anisotropy. The mode of anisotropy is useful for resolving whether a region of anisotropy is linear anisotropic, orthotropic, or planar anisotropic. Both tensor trace and fractional anisotropy are members of an orthogonal invariant set, but they do not belong to the same set. It is proven that tensor trace and fractional anisotropy are not mutually orthogonal measures of the diffusive process. The results are applied to the analysis and visualization of diffusion tensor magnetic resonance images of the brain in a healthy volunteer. The theoretical developments provide a method for generating scalar maps of the diffusion tensor data, including novel fractional anisotropy maps that are color encoded for the mode of anisotropy and directionally encoded colormaps of only linearly anisotropic structures, rather than of high fractional anisotropy structures.

Anisotropy↗

Experiments on movement of DNA regions in Escherichia coli evaluated by computer simulation.

During the cell cycle of Escherichia coli DNA is replicated and segregated over two prospective daughter cells. Nucleoids as a whole separate gradually in line with cell elongation, but sub-nucleoid DNA regions may behave differently, separating non-gradually. We tested the ability of three models to predict the outcome of a fluorescent in situ hybridisation (FISH) experiment. We did this by comparing computer-simulated data with experimental data. The first model predicts gradual separation in line with cell elongation. The second model predicts that origins stick together for some time after duplication before one copy jumps to the other side of the cell (non-gradual separation). The simulated data of these models are very similar, indicating that FISH is not a suitable method to distinguish between these two models. The third model predicts that origins may be anywhere within the nucleoid(s). We found that simulated data using the third model resemble the experimental data most. However, DNA regions are not randomly localised in the cell, although their localisation is fuzzy. We propose that movement of DNA regions is the result of a combination of factors. Nucleoid segregation (or the forces behind it) dictates the overall direction of movement. Other factors, of which we show that diffusion could be an important one, move DNA in other directions giving rise to non-gradual movement in individual cells and contributing to variation in intracellular position per cell length in a population of cells.

Cell Cycle↗

In situ study of the growth kinetics of individual island electrodeposition of copper.

The growth kinetics for individual islands during electrodeposition of copper have been studied using in situ transmission electron microscopy. We show that for sufficiently large overpotentials, the growth kinetics approach the rate laws expected for diffusion-limited growth of hemispherical islands, characterized by two distinct regimes. At short times, the island growth exponent is 0.5 as expected for diffusion-limited growth of uncoupled hemispherical islands, while at longer times, the growth exponent approaches 1/6 as expected for planar diffusion to the growing islands. These results provide the first direct measurements of the growth of individual islands during electrochemical deposition. However, quantitative comparison with rate laws shows that the island radii are smaller than predicted and the island densities are much larger than predicted, and we suggest that this is related to adatom formation and surface diffusion, processes which are not included in conventional growth models.

Journal Article↗

Tubulointerstitial immune complex nephritis in rats immunized with Tamm-Horsfall protein.

Rats immunized with Tamm-Horsfall protein (TH) developed autoantibodies to TH and tubulointerstitial nephritis with granular deposits of IgG, C3, and TH at the base of tubular cells of the thick ascending limb of Henle's loop and selective infiltration of leukocytes around this distal nephron segment. Selective IgG deposition at the base of cells in the tubular segment containing TH and the direct relationships of the amount of immune deposits and the severity of histologic lesions to serum levels of anti-TH antibodies suggest a pathogenetic role for the autoantibodies to TH. The immune deposits in this model of tubulointerstitial nephritis appear to be the consequence of in situ formation of immune complexes. This mechanism involves the diffusion of circulating antibodies to TH across tubular basement membranes and their combination with TH molecules associated with tubular cell surface membranes.

Animals↗

Electric-field-induced redistribution of polar nano-regions in a relaxor ferroelectric.

Relaxor ferroelectrics, with their strong dependence of polarization on the applied electric field, are of considerable technological importance. On a microscopic scale, however, there exists competition as well as coexistence between short-range and long-range polar order. The conventional picture is that the polar nano-regions (PNRs) that appear at high temperatures beyond the Curie transition, form nuclei for the field-induced long-range order at low temperatures. Here, we report high-energy X-ray diffuse-scattering measurements on the relaxor Pb(Zn(1/3)Nb(2/3))O(3) (PZN) to study the short-range polar order under an electric field applied along the [111] direction. In contrast to conventional expectations, the overall diffuse-scattering intensity is not suppressed. On the other hand, the field induces a marked change on the shape of the three-dimensional diffuse-scattering intensity pattern, corresponding to a redistribution of PNRs in real space. We show that these surprising results are consistent with a model in which the PNRs with [110]-type polarizations, orthogonal to that of the surrounding environment, are embedded and persist in the [111]-polarized ferroelectric order of the bulk.

Journal Article↗

Rational targeting for prion therapeutics.

Prions--pathogens that are lethal to humans and other animals--are thought to be conformational isomers of the cellular prion protein. Their unique biology, and the potential for a wider pathobiological significance of prion-like mechanisms, has motivated much research into understanding prion neurodegeneration. Moreover, concerns that extensive dietary exposure to bovine spongiform encephalopathy (BSE) prions might have infected many individuals--who might eventually develop its human counterpart, variant Creutzfeldt-Jakob disease (vCJD)--has focused much interest on therapeutics. The challenge of interrupting this aggressive, diffuse and uniformly fatal neurodegenerative process is daunting. However, the recent finding that the onset of clinical disease in established neuroinvasive prion infection in a mouse model can be halted and early pathology reversed is a source for considerable optimism. A therapeutic focus on the cellular prion protein, rather than prions themselves, which might not be directly neurotoxic, is suggested.

Animals↗

Mass transfers in a fluidized bed bioreactor using alginate beads for a future bioartificial liver.

Fluidized bed bioreactor with alginate beads may be an alternative to hollow fiber cartridge to host hepatocytes for bioartificial liver purposes. After the bioreactor design and the characterization of fluid mechanics, the present study was aimed at analyzing bi-directional mass transfers of calibrated species between external fluid and empty beads. Static (batch) and dynamic (fluidized bed bioreactor) experimental conditions were analyzed. A simple modelling approach permitted the definition of mass transfer coefficients. The motion of beads within the bioreactor clearly enhanced mass transfer kinetics, but did not alter the amount exchanged. The shear enhanced diffusion coefficient for VitB12 was 20 times higher in the fluidized bed bioreactor than under batch conditions, proving the efficiency of such a device.

Alginates↗

[Development of drug delivery system based on a new administration route for targeting to the specific region in the liver].

Development of drug delivery systems to achieve site-specific delivery or prolonged retention in the circulation has attracted attention, because new types of drugs are expected to be created with advances in life science and biotechnology such as the Human Genome Project. We have tried to develop a new administration route for drug targeting to the liver, since drug administration by the intravenous and oral routes makes it difficult to achieve a local site of action in the liver. Although direct application to the liver surface should result in local drug distribution, drug absorption from the liver surface has not been reported in the literature. Therefore we analyzed the absorption mechanism of several organic anions and dextrans with different molecular weights as model drugs, after application to the rat liver surface in vivo, employing a cylindrical diffusion cell. Every compound appeared gradually in the plasma, followed by excretion into the bile and/or urine, indicating the possibility of drug absorption from the liver surface. A specific transport system might not be involved in the absorption process from the liver surface, because the effect of dose and transport inhibitors on the absorption was not recognized. In addition, molecular weight was found to be a determining factor in absorption from the liver surface. The targeting efficacy was considerably enhanced by application to the liver surface, as compared with intravenous administration. Moreover, we have identified important physicochemical and pharmaceutical factors determining the absorption rate of a drug from the liver surface for clinical use. Consequently, drug application to the liver surface could improve availability in the desired site of a new drug such as bioactive compounds and genomic medicines, by combination with appropriate chemical and pharmaceutical formulation modifications.

Animals↗