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In this study, a very dilute solution (NH(4)OH:H(2)O(2):H(2)O 1:8:64 mixture) was employed to reduce the thickness of commercially available SOI wafers down to 3 nm. The etch rate is precisely controlled at 0.11 Å s(-1) based on the self-limited etching speed of the solution. The thickness uniformity of the thin film, evaluated by spectroscopic ellipsometry and by high-resolution x-ray reflectivity, remains constant through the thinning process. Moreover, the film roughness, analyzed by atomic force microscopy, slightly improves during the thinning process. The residual stress in the thin film is much smaller than that obtained by sacrificial oxidation. Mobility, measured by means of a bridge-type Hall bar on 15 nm film, is not significantly reduced compared to the value of bulk silicon. Finally, the thinned SOI wafers were used to fabricate Schottky-barrier metal-oxide-semiconductor field-effect transistors with a gate length down to 30 nm, featuring state-of-the-art current drive performance.

The elastic properties of nanotubes fabricated by layer-by-layer (LbL) assembly of polyelectrolytes in the nanopores of polycarbonate track-etched membranes have been investigated by resonant contact Atomic Force Microscopy (AFM), for nanotube diameters in the range of 100 to 200 nm. The elastic modulus of the nanotubes was computed from the resonance frequencies of a cantilever resting on freely suspended LbL nanotubes. An average value of 115 MPa was found in air for Young's modulus of these nanostructures, well below the values reported for dry, flat multilayers, but in the range of values reported for water-swollen flat multilayers. These low values are most probably due to the lower degree of ionic cross-linking of LbL nanotubes and their consequently higher water content in air, resulting from the peculiar mode of growth of nanoconfined polyelectrolyte multilayers.

We report on the grafting of coumarin chromophores on flat silicon surfaces and in regions of nanometric dimensions drawn on silicon surfaces. The coumarin derivative was grafted by using the quaternization of a tertiary amine group of the chromophore with a ((chloromethyl)phenylethyl)-dimethylchlorosilane (CMPDCS) grafted on silicon. Complete characterization of the grafted layer was performed as a function of reaction time by X-ray photoelectron spectroscopy, X-ray reflectometry, atomic force microscopy, fluorescence spectroscopy and laser-scanning confocal microscopy. The results indicate that about one chromophore molecule is grafted every second CMPDCS molecule, resulting in a surface density of coumarin of slightly more than one coumarin per nm2. A broadening of the distribution of the fluorescence lifetimes was observed, suggesting that the grafted molecules experience a larger distribution of environments in the grafted layer than in solution. Since this reaction is fully compatible with silicon processing technology, the grafting could also be performed in nano-regions of size as small as 250 nm defined by combining electron-beam lithography with silanization. In such nano-sized regions the distribution of fluorescence lifetimes was narrower, suggesting a possible influence of the confinement on the organization of the molecules.

We demonstrate that arrays of nanowires of conjugated polymers can be easily produced by a simple embossing protocol, compatible with very large scale integration technology. The embossing process is shown to have the supplementary virtue to increase the internal degree of order of the nanowires, significantly enhancing their performance. This is applied to the fabrication of nanowire-based devices consisting of a liquid crystalline light-emitting polymer, of a liquid crystalline semiconducting polymer, and of an amorphous conducting polymer, illustrating the versatility and wide applicability of the method.

The ability of poly(ethylene oxide)-silane (PEO-silane) monolayers grafted onto silicon surfaces to resist the growth of polyelectrolyte multilayers under various pH conditions is assessed for different pairs of polyelectrolytes of varying molar mass. For acidic conditions (pH 3), the PEO-silane monolayers exhibit good polyelectrolyte repellency provided the polyelectrolytes bear no moieties that are able to form hydrogen bonds with the ether groups of the PEO chains. At basic pH, PEO-silane monolayers undergo substantial hydrolysis leading to the formation of negatively charged defects in the monolayers, which then play the role of adsorption sites for the polycation. Once the polycation is adsorbed, multilayer growth ensues. Because this is defect-driven growth, the multilayer is not continuous and is made of blobs or an open network of adsorbed strands. For such conditions, the molar mass of the polyelectrolyte plays a key role, with polyelectrolyte chains of larger molar mass adsorbing on a larger number of defects, resulting in stronger anchoring of the polyelectrolyte complex on the surfaces and faster subsequent growth of the multilayer. For polyelectrolytes of sufficiently low molar mass at pH 9, the growth of the multilayer can nevertheless be prevented for as much as five cycles of deposition.

By confining discotic phthalocyanines in a network of crisscrossed nanogrooves, we obtain a uniaxial alignment of the columnar mesophase. The alignment process is based on the anisotropy of interface tension between the mesophase and the nanogrooves' walls. Preferential mesophase alignment results from this nonhomogeneity combined with the anisotropy of the network cell dimensions. A simple model is proposed to explain the experimental observations.

With the purpose of developing biosensors, the reliable proof of the biological activity of two new sensor systems was obtained by atomic force microscopy (AFM) in both the imaging and the single-molecule force spectroscopy modes. Antigens or antibodies of pharmacological interest were grafted onto self-assembled monolayers of thiols on gold, and AFM imaging demonstrated that the grafting process produced homogeneous submonolayers of isolated proteins. The analysis of the morphology of the surfaces at the different functionalization steps allowed evaluating the protein grafting density and showed that the recognition of complementary species present in the surrounding solution occurred. Single-molecule force spectroscopy experiments between the sensing surfaces and AFM probes, onto which the complementary species were grafted, enabled a direct and rapid test of the biological activity of the sensors by investigating the interaction occurring at the level of one single ligand-receptor bond. Ellipsometry and surface plasmon resonance allowed further characterization of the sensor surfaces and confirmed that the biological recognition took place.

Spherical micellar aggregates have been obtained in chloroform by mixing poly(styrene)-block-poly(4-vinylpyridine) (PS-b-P4VP) diblock copolymers with perfluorinated surfactants (FS) bearing a carboxylic acid head. These micellar aggregates are resulting from the self-assembly of the insoluble P4VP/fluorinated complexes into a core surrounded by the soluble PS coronal chains. Their characteristic features have been studied as a function of various parameters including the composition of the PS-b-P4VP copolymer, the tail length of the fluorinated surfactant, the 4VP/FS molar ratio, the number of carboxylic acid group (1 or 2) on the surfactant, the presence of the PS block and of the fluorine atoms on the surfactant. Dilution of these initial micellar aggregates triggers a morphological reorganization resulting in the formation of more stable vesicles. The extent of this morphological transition is related to the solubility of the P4VP/fluorinated complexes during the dilution process. This transition is complete for short P4VP/FS complexes, incomplete for long P4VP/FS complexes, and not observed whenever an alpha,omega-difunctional FS is used in P4VP/FS complexes, leading to a cross-linked core. Finally, the spheres-to-vesicles transition has been advantageously used in order to encapsulate molecules, as demonstrated by confocal fluorescence microscopy.

The formation of micelles on surfaces by spin-coating dilute solutions of diblock, triblock, and tetrablock copoly(2-oxazoline)s in a non-selective solvent is demonstrated. The micelles are not preexistent in the initial solution but are formed during the evaporation of the solvent by precipitation of the least-soluble block. The morphology and size of the micelles vary according to the fraction of this block but are not dependent on the block order in the copolymer.

Recently, we reported on "self-rupturing" microcapsules which consist of a biodegradable dextran-based microgel surrounded by a polyelectrolyte membrane. Degradation of the microgel increases the swelling pressure in the microcapsules which, when sufficiently high, ruptures the surrounding polyelectrolyte membrane. The membrane surrounding the microgels is deposited using the layer-by-layer (LbL) technique, which is based on the alternate adsorption of oppositely charged polyelectrolytes onto a charged substrate. In this paper, we characterize with confocal microscopy, electrophoretic mobility, scanning electron microscopy and atomic force microscopy in detail the deposition and the properties of the LbL coatings on the dextran microgels. We show that by fine-tuning the properties of both the microgel core and the LbL membrane the swelling pressure which is evoked by the degradation of the microgel is indeed able to rupture the surrounding LbL membrane. Further, we show that the application of an LbL coating on the surface of the microgels dramatically lowers the burst release from the microcapsules and results in massive release at the time the microcapsules rupture.

Phenylboronic acids are known to form covalent complexes with polyol compounds such as glucose. A novel polyelectrolyte, containing phenylboronic acid as a glucose-sensitive moiety, has been synthesized and used for the fabrication of glucose-sensitive hollow polyelectrolyte capsules using the layer-by-layer technique. The response to glucose was observed as a rather fast dissolution of the capsules when brought into contact with a glucose-containing medium. These polyelectrolyte capsules are the first polyelectrolyte capsules able to respond to a stimulus that can be provided by the human body (i.e., an increase in glucose concentration). Therefore, the concept we present has promising applications in the biomedical field for the controlled delivery of insulin.

Microwave-assisted polymerization has been utilized to synthesize amphiphilic poly(2-ethyl-2-oxazoline-block-2-"soy alkyl"-2-oxazoline) diblock copolymers (PEtOx-PSoyOx). The amphiphilic block copolymers have been used to prepare aqueous spherical micelles consisting of a PEtOx corona and a PSoyOx core, which have been further cross-linked by UV irradiation. The morphology of these cross-linked micelles has been shown to reversibly change from spheres to short rods referred to as rice grains whenever the micelles were transferred from water into acetone, a nonselective solvent for the constituent blocks. This morphological transition has been attributed to the swelling of the slightly cross-linked PSoyOx core.

We report on the fabrication of chemically nanopatterned gold surfaces by combining electron-beam lithography with gas and liquid phase thiolization. The line-edge roughness of the patterns is ∼4 nm, corresponding to a limiting feature size in the range of 15 nm. Indications for a lower packing density of the self-assembled monolayers grown in the nanofeatures are given, and evidences for the bleeding of thiols along the grain boundaries of the gold substrate are displayed. A comparison is provided between nanopatterned thiol and silane monolayers on gold and on silicon wafers, respectively. The line-edge roughnesses are shown to be close to each other for these two systems, indicating that the limiting step is currently the lithography step, suggesting possible improvement of the resolution. The advantages and drawbacks of thiol versus silane monolayers are finally discussed with respect to the formation of chemically nanopatterned surfaces.

The control and modification of surface state is a major challenge in bioanalytical sciences, and in particular in electrokinetic separation methods, due to the importance of electroosmosis. This topic has gained recently a renewed interest, associated with the development of "lab-on-chips" systems that extend the range of materials in which separation channels are fabricated. The surface science community has developed through the years a large toolbox of characterization tools and surface modification protocols, which is not yet fully exploited in the bioanalytical world. In this paper, we try and present an overview of these tools, in order to stimulate new ideas for improved and more controlled surface treatment strategies for separations in capillaries and microchannels. We briefly describe some physical and chemical aspects of electroosmosis (global and spatially resolved), streaming current, and streaming potential. We also review the main strategies for surface coating, and compare the advantages of physisorption, well-organized thin self-assembled monolayers, or conversely thick polymer "brushes". Examples of existing applications to electrophoresis in microchannel are also given.

We compare two routes for creating protein adsorption-resistant self-assembled monolayers (SAMs) by chemical modification of silicon surfaces with poly(ethylene oxide) (PEO) oligomeric derivatives. The first route involves the assembly of 2-methyl[(polyethyleneoxy)propyl]trichlorosilane (Cl3SiMPEO) films onto oxidized silicon surfaces (OH-SiO(x)) either by a liquid-phase process at room temperature or by a gas-phase process at 423 K, producing Si-O-Si bonds between the substrate and the organic layer. The second pathway makes use of the assembly of poly(ethylene glycol methyl ether) (MPEG) films onto hydrogen-passivated silicon surfaces (H-Si) using a liquid-phase process at 353 or 423 K, leading to the formation of Si-O-C bonds between the substrate and the organic layer. Structural investigation by X-ray reflectometry (XRR) reveals that the thickness and surface densities of the grafted PEO monolayers strongly depend on experimental conditions such as temperature and grafting time. Atomic force microscopy (AFM) shows that very smooth and homogeneous monolayers can be obtained with average roughnesses close to those measured on the corresponding bare substrates. Finally, the antifouling properties of the modified silicon surfaces were evaluated by X-ray photoelectron spectroscopy (XPS), using a membrane protein (P.69 antigen) as model protein. Both types of PEO monolayers exhibit excellent protein repellency, as soon as the grafting density is equal to or higher than 1.7 chains/nm2.

Self-exploding microparticles show potential for advanced delivery of certain therapeutics. This study evaluates (1) whether degrading biodegradable dextran hydroxyethyl methacrylate (dex-HEMA) microgels can be coated by a lipid membrane and (2) whether the surrounding membrane can be ruptured by the increasing swelling pressure of the degrading microgel. We found that adsorption of charged liposomes to oppositely charged dex-HEMA microgels provides efficient coating of the microgels, whereby microparticles with a "core-shell" structure were clearly obtained. Especially, we could confirm experimentally that the swelling pressure increase of degrading dex-HEMA microgels can destroy the lipid membrane surrounding the microgels.

Collagen, a protein widely used to control cell-material interactions, is known to self-assemble in solution. Supramolecular structures also form on material surfaces following collagen adsorption. Herein, we report the use of anisotropic, flat, surface chemical nanopatterns, which consist of alkyl-terminated tracks drawn in an oligo(ethylene glycol)-terminated matrix, to direct collagen adsorption. As revealed by atomic force microscopy, the spontaneous collagen adsorption performed on such patterned substrates results in the accumulation of collagen on the hydrophobic tracks. Moreover, the width of the tracks (30-90 nm), which is much smaller than the length of the collagen molecule (approximately 300 nm), is the origin of preferential alignment of the molecules and of their assembly into continuous bundles of adsorbed collagen. This chemical guidance effect due to self-confinement of proteins upon adsorption may bring novel and valuable applications, specifically in biomaterials science and cell growth control.

Polymer crystallization is notoriously difficult to control. Here, we demonstrate that the orientation of polymer crystals can be fully controlled at the nanoscale by using nanoimprint lithography (NIL) with molds bearing nanotrenches to shape thin films of poly(vinylidene fluoride). This unprecedented control is due to the thermomechanical history experienced by the polymer during embossing, to the shift of the nucleation mechanism from heterogeneous to homogeneous in confined regions of the mold, and to the constraining of the fast growth axis along the direction of the trenches. NIL thus appears as an ideal tool to realize smart polymer surfaces where crystal ordering can be tuned locally.

The effect of roughness on the dewetting behavior of polyethylene thin films on silicon dioxide substrates is presented. Smooth and rough silicon dioxide substrates of 0.3 and 3.2-3.9 nm root-mean-square roughness were prepared by thermal oxidation of silicon wafers and plasma-enhanced chemical vapor deposition on silicon wafers, respectively. Polymer thin films of approximately 80 nm thickness were deposited by spin-coating on these substrates. Subsequent dewetting and crystallization of the polyethylene were observed by hot-stage optical microscopy in reflection mode. During heating, the polymer films melt and dewet on both substrates. Further observations after cooling indicate that, whereas complete dewetting occurs on the smooth substrate surface, partial dewetting occurs for the polymer film on the rough surface. The average thickness of the residual film on the rough surface was determined by ellipsometry to be a few nanometers, and the spatial distribution of the polymer in the cavities of the rough surface could be obtained by X-ray reflectometry. The residual film originates from the impregnation of the porous surface by the polymer fluid, leading to the observed partial dewetting behavior. This new type of partial dewetting should have important practical consequences, as most real surfaces exhibit significant roughness.

We report on the exchange between a hydrophilic thiol (11-mercapto-1-undecanol) in a liquid or gas phase and a hydrophobic thiol (dodecanethiol) of similar length self-assembled on a polycrystalline gold surface for a wide range of temperatures and times. The molecular composition of the mixed monolayers is determined by the static water contact angle and X-ray photoelectron spectroscopy measurements. Atomic force microscopy in lateral force mode is used to characterize the molecular domains at the nanometer level. The exchange first occurs rapidly at the gold grain boundaries, with an activation energy of about 66 +/- 4 kJ/mol. Then, boundaries of ordered thiol domains are progressively replaced, and the exchange is slowed because only regions of increasing perfection are left untouched. Higher temperatures lead to faster kinetics of replacement and the removal of larger amounts of the original thiol. No significant difference could be detected between exchange occurring in an ethanol solution or in the gas phase, and the initial rate of exchange was found to be similar for the displacement of dodecanethiol by 11-mercapto-1-undecanol molecules and for the converse displacement.

Metastable spherical micelles have been obtained by mixing in chloroform poly(styrene)-block-poly(4-vinylpyridine) diblock copolymers with perfluorinated surfactants bearing a carboxylic acid head. Dilution of these initial micelles triggers a morphological reorganization resulting in the formation of more stable vesicles. This transition can be advantageously used to encapsulate molecules of interest.

We investigate the adsorption of a globular protein (P.69 pertactin, also known as antigen 69k) on protein-repellent hydrophilic substrates bearing regularly spaced hydrophobic nanostripes, for stripe widths comprised between 20 and 160 nm. Protein adsorption is shown to be remarkably well-controlled by the templating substrates, with a near-to-perfect reproduction of stripes by the protein monolayer down to 20 nm width, except for a 5-10 nm broadening. However, whereas the ellipsoidal protein forms a dense monolayer with random orientation of its long axis for large stripe widths, it adsorbs in a predominantly side-on (flat-on) orientation for stripe widths below 50 nm, due to the easier reorientation (interfacial relaxation) of the proteins adsorbed at the edges of the stripes, which experience a decreased lateral interaction. These results show that protein confinement in regions of a size similar to their dimensions can be used to tune their orientation, which may be of interest for applications in high-density sensor devices.

We present a new way to protect polyelectrolyte multilayers from water, consisting in the adsorption and subsequent fusing of charged wax particles atop a multilayer. The formation of the wax layer is demonstrated by different techniques such as ellipsometry, contact angle measurements, and atomic force microscopy. The diffusion of water in protected and unprotected multilayers is studied by in situ neutron reflectometry. Whereas a top layer of wax crystals already allows substantial reduction of the diffusion, the fusion of this top layer leads to the dominating exclusion of water from the multilayers when dipped in water. This method opens up new interesting avenues for polyelectrolyte multilayers in practical applications where permeability of water, ions, or hydrophilic drugs is an issue.

The development of robust methodologies to control the solid-state structure of polymeric materials by appropriate design of the macromolecular architecture has a crucial impact on the mechanical properties of these materials. Here, we demonstrate the feasibility of controlling chain folding of polymers by steric interactions only, in contrast to previous attempts aimed at engineering polymer crystallization through hydrogen bonding. In a linear synthetic macromolecule similar to polyethylene, we encoded structural instructions that are translated during a crystallization process to generate a unique, semi-crystalline morphology with structure-controlled crystal thickness of approximately 5 nm that remains constant over a wide temperature range. The molecular code consists of a linear backbone alternating crystallizable, long alkyl sequences of monodisperse sizes separated by short spacers containing side-chains and acting as stops and fold-controlling units. This simple strategy could be used to produce advanced polymeric materials with fine control of the crystalline and amorphous regions.

Energetic considerations indicate that long-range Van der Waals forces stabilize thin polystyrene (PS) films against height fluctuations on silicon substrates. Nevertheless, we report here on the amplification of capillary waves of specific wavelengths for 15 nm thick PS films on silicon, ultimately leading to dewetting in a "spinodal-like" process. However, the temporal dependence of the wavelength of the growing instability does not agree with the "classical" spinodal dewetting mechanism. Therefore, this phenomenon is ascribed to the existence of "structural" forces resulting either from the restructuring of the films or from density variations within the films during annealing, in accordance with recent theoretical treatments. The process is shown not to be limited to polystyrene films, which indicates the generality of our findings.

We study the growth and internal structure of polyelectrolyte multilayers obtained by combining three polyanions with nine polycations of the ionene family, of systematically varied chemical architecture. We find that, contrary to a generally held belief, ordered organic multilayers are by no way exceptional, provided one of the polyelectrolytes bears groups which induce structure in water, such as long hydrophobic segments or mesogenic groups. However, this condition is not sufficient, as order will or will not emerge in the multilayer depending on the specific pairing of the polyelectrolytes. The results support the notion that layering in the multilayer results from some degree of prestructuring of a water-swollen layer adsorbed during each step of deposition. These findings pave the way to new possible uses of polyelectrolyte multilayers, for example, for applications requiring preferential alignment or strong confinement of specific functional groups.

The consequences of profound early deafness on oral language in children are drastic. The modern cochlear implant (CI) has been shown to enhance speech production skills in prelingually deaf children. Many factors may contribute to a poor or an excellent outcome, making it difficult to compare groups of children wearing or not wearing cochlear implants. The present study compared receptive language levels in matched pairs of children from CI group and non-CI groups. The pre-op receptive language development curve suggest a possible growth over time with the maturation and the speech therapy. Comparison showed that the slope for post-op CI children to be greater than for non-CI children, and that this difference is statistically significant, and that the slope for CI children to be greater post- than pre-operatively. The main conclusion is that receptive language scores grow significantly higher over time after surgery in CI than in pair-matched non-CI children, despite better initial pure tone audiometric thresholds of the latter.

The consequences of profound deafness on oral language development in children are drastic and well-known. Modern multichannel cochlear implant (CI) has been proven to enhance speech production skills in prelingually deaf children. Speech production skills, however, are known not to be a reliable reflection of oral language competence as a whole. Language is an acquired common code in a specific group, enabling exchange of ideas, feelings and knowledge. In humans, speech is one of the channels conveying language. Assessing language development in CI children is more difficult than simply assessing speech production skills. Many factors may contribute to a poor or an excellent outcome, making it difficult to compare groups of children wearing or not wearing CI. The present study compared receptive language levels in paired matched children from CI and non-CI groups. The main conclusion of this study is that language comprehension scores grow significantly higher over time post-surgery in CI than in paired-matched non-CI children, despite better initial pure tone audiometric thresholds of the latter.

Ketamine hydrochloride, used during glucose or lactose tolerance tests to aid restraint of rhesus and stumptail macaques unconditioned to handling, prevented the establishment of definitive baseline plasma glucose tolerance curves from which meaningful interpretations could be derived. Repetition of the tests after the animals were restraint-chair conditioned provided clear baseline control data. It is suggested that ketamine might invalidate those carbohydrate tolerance tests which are based on the absorption of glucose from the gastrointestinal tract.

The etiology of a transmissable colonic mucosal hyperplasia of mice was investigated. Hyperplasia was produced in mice inoculated with unfiltered colonic suspension from affected mice, but infectivity was lost after passage through a 0.45 mum filter. The etiologic agent was subsequently identified as a variant of Citrobacter freundii. The organism induced colonic mucosal hyperplasia when inoculated into germfree mice, and it was recovered in pure culture from the affected animals.

A health assessment procurement program should be well matched with an ongoing in-house health monitoring and evaluation program. Programs for health assessment include ordering, receiving and quarantining of animals, or the placement of vendor animals directly into colony rooms. Health assessment methods for procured laboratory rodents consist of statistical sampling, clinical evaluation, laboratory evaluation, reporting results to veterinarians, interpretation of data, communication to investigators and vendors and action.

The morphogenesis of 1, 2-dimethylhydrazine (DMH)-induced lesions in the colon of outbred NIH Swiss mice was determined for up to 5 months of treatment. The effect of hyperplasia on DMH carcinogenesis was also evaluated by introducing a transient hyperplastic stimulus to the colon during the chronic weekly treatment regimen of DMH. The hyperplastic stimulus was a naturally occurring disease of mice, transmissible murine colonic hyperplasia, which is caused by a variant of Citrobacter freundii. In control mice, those not receiving the bacterium, weekly injections of the carcinogen induced neoplastic changes first detectable at two months of treatment in all segments of the colon and in both sexes. The changes increased in frequency and severity with time. Diffuse mucosal hyperpladia and chronic inflammatory and degenerative changes were also associated with DMH after prolonged treatment. The hyperplastic stimulus of C. freundii reduced the latent period for appearance of early DMH tumors, but it had no influence on already established DMH tumors.

Conditions for oral transmission of atypical ileal hyperplasia (AIH) in weanling hamsters were established and 22 passages were made. AIH was transmitted by feeding whole cell-free supernatants of ileal homogenates. The etiologic agent(s) was retained by 0.22 mum pore-size filters and was inactivated by chloroform treatment or by heating at 56 degrees C for 30 min. Enteric bacteria from affected animals also induced AIH, but with a lower morbidity and mortality than following inoculation with ileal extracts. Experimentally induced lesions progressed from marked segmental hyperplasia of ileal mucosa to granulomatous inflammation in underlying connective tissue and muscle tunics. Hyperplastic mucosal epithelium penetrated the muscularis mucosa, but metastases were not detected. Serum antibody from exposed animals reacted specifically, by indirect immunofluorescence, with an intracytoplasmic mucosal cell antigen(s) of autologous and homogolous ileal lesions, but antibody did not react with normal ileal mucosa or with unaffected portions of intestine from animals bearing ileal lesions.

The toxic effects of cytembena in beagle dogs and rhesus monkeys were investigated with the drug given as single or daily iv injections in doses ranging from 12.5 to 200 mg/kg/day to dogs and 6.25 to 50 mg/kg/day to monkeys. Renal tubular damage was a major drug- and dose-related finding in both species and was clinically indicated by an accompanying uremia, elevated serum creatinine, and proteinuria. In the kidney, the primary lesion was cellular necrosis and desquamation of the distal tubular epithelium in animals given the lowest toxic doses. More severe but similar histologic changes produced by this drug were further characterized by single dose studies in mice which showed renal mitochondrial swelling and disruption plus generalized cell swelling as progressive, subcellular developments which were well established 24 hours after treatment. Cellular regeneration in the renal tubular epithelium was found in dogs and monkeys retained 6 weeks for observation after treatment, although functional recovery was inconsistent. A toxic effect to lymphoid tissue was an additional finding which is described.

A lethal disease characterized by hemorrhage and necrosis of the brain, testes, and epididymides developed in young adult rats housed in specific pathogen free quarters. Morphological, virological, and serological investigations of the outbreak indicated that the probable causative agent was rat virus (Kilham), a common parvovirus of rats that usually induces persistent, asymptomatic infection in adult rats.

Sialodacryoadenitis virus (SDAV), a coronavirus of rats, evoked both serum neutralization and complement fixation antibody responses when inoculated intranasally in mice. Weanling gnotobiotic CD-1 mice inoculated intranasally with 10(3.0) mean tissue culture infective doses of SDAV remained asymptomatic. Virus was recovered from the nasopharynx, trachea, and lung from day 2 to day 7. Viral antigen was readily detected by indirect immunofluorescence in the lung but rarely in the nasopharynx. Infected mice developed interstitial pneumonia. Susceptible mice contact exposed to experimentally infected mice developed antibody to SDAV. Epizootiological studies indicated that retired breeder mice can have complement-fixing antibody to SDAV and mouse hepatitis virus (MHV) in the absence of MHV infection. These studies show that SDAV is infectious for mice and can be a pathogen for the respiratory system. Thus, SDAV infection of mice may be responsible for spurious seroconversions to MHV.

Pathology, microbiology, and selected serum chemistries were evaluated in 144 male Fischer rats from 4 to 33 mo of age. The rats were reared and maintained under barrier conditions, which successfully excluded the introduction of major infectious disease agents throughout the entire study, including Mycoplasma pulmonis. A wide variety of pathology was found and tabulated, and many lesions were found to increase in severity and incidence with age. There was a high correlation of renal disease severity with increasing age, while alpha-1 globulin and cholesterol increased.

Transmissible murine colonic hyperplasia, cuased by a variant of Citrobacter freundii (4280). was shown to be modified by diet and by host strain and species. Four different diets fed to mice inoculated with C frundii 4280 were found to have a significant but varying influence on the severity of hyperplasia. Diet also influenced the colonic crypt height of uninoculated, control mice. F344 rats, Syrian hamsters, and NIH Swiss [N:(S)], C57BL/6J, C3H/HeJ, and DBA/2J mice were inoculated with C freundii 4280. Marked strain differences were noted in the mice in mortality and severity of the colonic hyperplasia. The NIH Swiss mice had the greatest and the C57BL/6J mice had the least mucosal hyperplasia. The rats and hamsters did not develop disease or maintain infection after inoculation with the organism. Twenty isolates of Citrobacter from a range of biologic sources were inoculated into susceptible mice, but only mice inoculated with C freundii 4280 developed the disease.

The criteria for the selection of stocks and strains of laboratory animals were discussed. These include metabolic and environmental factors as well as the presence or introduction of infectious agents. The investigator is now concerned with his experimental animals from conception to death, not just during an experiment. The basis of a health monitoring program to ascertain the health status of incoming animals, to evaluate the status of the environment, and to document the ongoing status of an animal colony was outlined. The facilities of the Yale School of Medicine animal research space were detailed.

Morphological changes in retinas of aging Fischer 344 rats were characterized. The numbers of photoreceptor cells gradually decreased as rats aged. The outer nuclear layer was 12 cells thick at 3 months, but was reduced to less than 8 cells by 18 months. The decrease of photoreceptor cells was more pronounced in rats housed under a light intensity of 32-ft-c than in rats housed under a light intensity of 1 ft-c. Inner and outer segments of surviving photoreceptor cells were morphologically normal. A new form of retinal degeneration was discovered in aged Fischer rats characterized by selective degeneration of peripheral retina. Degeneration was characterized by severe loss of photoreceptor cells in the far peripheral retina. Microcystoids were found in about 25% ofthe affected retinas, and the loss of photoreceptor cells was followed by proliferation and vascularization of the retinal pigment epithelium and disorganization of retinal structures. The incidence and severity of peripheral retinal degeneration increased with aged and prolonged exposure to comparatively high-intensity light. All Fischer rats ((5/5) housed under light intensity of 32 ft-c developed severe peripheral retinal degeneration by 24 months. Peripheral retinal degeneration was an age-related change but appeared to be exaggerated by ambient light.

The disorder in Wag/Rij rats is a spontaneous, bilateral retinal deneration. It is characterized by an early onset, slowly progressive degeneration of the photoreceptor cells leading to destruction of the retina. Degeneration affects both rod cells and cone cells, and to a lesser degree the cells in the inner nuclear layer. The remarkable alterations in the retinal pigment epithelium during the course of the disease suggest a profound change in metabolism and function of the pigment epithelium and implicate a possibility of interaction between the pigment epithelium and the photoreceptors. Since degenerated cells have also been observed in the inner nuclear layers, there is a possibility that Muller's cells are involved in the retinal degeneration. Controlled experiments have demonstrated that the disease is not induced by light damaging effects of the retina, and initial breeding experiments suggest that the disease is inheritable, probably as an autosomal dominant trait. The retinal degeneration in Wag/Rij rats is a new, unique system and it is a potentially very useful animal model of retinitis pigmentosa.