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PubMed1985

On the heterogeneity of Purkinje neurons in vertebrates. Cytochemical and morphological studies of chromatin during eel (Anguilla anguilla L.) life cycle.

The relationship between the heterogeneity of Purkinje cell chromatin and the functional involvement of cerebellum has been analyzed comparing two stages of eel life cycle (yellow eel and silver eel) which are characterized by a different degree of swimming activity. The DNA content and the chromatin condensation have been studied by means of microdensitometry (Feulgen-DNA, Feulgen-DNA/area), microfluorometry (DNA after intercalation with Propidium Iodide at low and high concentration) and electron microscopy. In the transition from the yellow eel to the silver eel, the Purkinje cell heterogeneity undergoes variations. In the silver eel, which shows a higher swimming activity, high Feulgen-DNA and Propidium Iodide-DNA (H2c to 4c) were more frequent and a greater heterogeneity in the chromatin condensation was observed. In particular, the changes were more evident in the lobus vestibulolateralis; mainly in this area, which receives inputs from the lateral line, the Purkinje cells are scattered in the molecular layer. On the contrary, in the yellow eel these large cells occupy the zone near or within the granular layer. The data suggest a relationship between the degree of heterogeneity of Purkinje cells and the afferent systems of the cerebellar areas examined. The changes of cerebellar cytoarchitectonics may be expression of the higher degree of the swimming activity of silver eel.

Anguilla↗
PubMed2001

Anatomy and histology of the brain and sense organs of the Antarctic eel cod Muraenolepis microps (Gadiformes; Muraenolepididae).

Brain regions, cranial nerves, and sense organs in Muraenolepis microps, an Antarctic gadiform fish, were examined to determine which features could be attributed to a gadiform ancestry and which to habitation of Antarctic waters. We found that the central nervous system and sense organs are well developed, showing neither substantial regression nor hypertrophy. A detailed drawing of the brain and cranial nerves is provided. The rostral position of the olfactory bulbs and telencephalic size and lobation are common for the order. The optic tectum and corpus cerebelli are smaller than in most other gadiforms. The shape of the corpus cerebelli is not distinctive among gadiforms. The lateral line region is moderately well-developed, but not hypertrophied to the extent seen in deep-sea gadiforms. As is the case in gadids possessing barbels and elongated pelvic rays, Muraenolepis has well-developed facial lobes, although these are smaller and more laterally positioned. The vagal lobes are deeply placed in the rhombencephalon and project into the fourth ventricle. The brain of Muraenolepis resembles that of a phyletically derived gadoid, especially a phycid, more than it resembles the brain of a phyletically basal macrourid. Two histological features of the diencephalon of Muraenolepis appear to be unique among gadiforms: a well-organized thalamic central medial nucleus and subependymal expansions. Muraenolepis has a pure rod retina like many deep-sea species but lacks the superimposed layers of rod outer segments. The histology of the nonvisual sense organs, especially the olfactory and external taste systems, are well-developed in Muraenolepis but not hypertrophied. We relate our findings to what is known about neural morphology in other gadiforms and in phyletically distant notothenioids and liparids that are sympatric with Muraenolepis on the Antarctic shelf. The only feature that reflects an Antarctic existence is the diencephalic subependymal expansions, which within notothenioids mirror the habitation of cold waters and have been found in every Antarctic species examined to date. Although the waters of the Antarctic shelf are cold, dark, and deep, brain and sense organ morphology in Muraenolepis are remarkably free of extreme specialization.

Animals↗
PubMed1994

Intrapelvic pressure monitoring in the partially obstructed porcine kidney.

OBJECTIVES: The diagnosis of urinary obstruction in newborns and infants remains difficult because the diagnostic studies available at present are fraught with many problems. It is our premise that precise measurement of renal pelvic pressures under physiologic conditions in patients with urinary tract dilation will allow diagnosis of obstruction and prediction of further renal damage. This study evaluates an intrapelvic pressure monitoring system. METHODS: Four porcine subjects had partial ureteropelvic junction obstruction created surgically; two subjects were used as controls. Doppler ultrasound and MAG-3 furosemide renography were performed preoperatively and each week pressures were measured with a Millar 5 F solid-state pressure transducer and the analog signals were converted on line to digital information to allow signal processing and later data analysis. Antegrade nephrostograms were obtained at the same sitting and showed that the model remained stable over time. Measurements were obtained at physiologic flow rates as well as at the standard Whitaker infusion rate of 10 cc/min. RESULTS: Thirteen pressure studies were performed on 9 kidneys. Each study was classified as normal (4), partial obstruction (8), or total obstruction (1), based on ultrasound and nephrostogram criteria. The normal units had baseline pressures of 4.12 +/- 0.94 cm water (H2O), which increased to 9.12 +/- 1.38 cm H2O with infusion. In the partially obstructed group, baseline was 16.4 +/- 3.83 cm H2O and increased to 35.3 +/- 15.9 cm H2O with infusion. The kidney with total obstruction showed a baseline pressure of 27 cm H2O, which increased to 68 cm H2O with infusion. These pressure measurements were also compared to furosemide renography. CONCLUSIONS: Intrapelvic pressures correlated well with conventional ultrasound and nephrostogram in the evaluation of partial obstruction. However, the results of furosemide renography and the Whitaker test were quite variable and did not accurately define partial obstruction or correlate with intrapelvic pressure. The Millar solid-state transducer monitoring system is an accurate method of evaluating the intrapelvic pressure and could possibly become a standard for diagnosing significant urinary obstruction against which other studies can be evaluated.

Animals↗
PubMed2000

Circulating levels of tumour necrosis factor-alpha and its soluble receptors are increased in the blood of patients with amyotrophic lateral sclerosis.

An immunologic pathogenesis for amyotrophic lateral sclerosis (ALS) has been recently proposed. We tested the whole tumour necrosis factor (TNF) system in the serum of 51 ALS patients at different stages of the disease and 36 healthy controls. Antigenic TNF-alpha and its soluble receptors (sTNF-Rs), measured by ELISA, were significantly higher in ALS patients than in healthy controls. However, biologically active TNF-alpha, corresponding to the sTNF-Rs-unbound trimeric TNFalpha molecule and assayed by its cytotoxic activity on a sensitive cell line, was similar between ALS patients and healthy controls. Neither antigenic TNF-alpha, bioactive TNF-alpha nor sTNF-Rs correlated with disease severity, disease duration, or weight loss. In conclusion, we reported an activation of the TNF system in ALS. The role of this activation in the pathogenesis of the disease remains elusive.

Amyotrophic Lateral Sclerosis↗
PubMed2006

Molecular and cytogenetic changes involved in the immortalization of nasopharyngeal epithelial cells by telomerase.

Nasopharyngeal carcinoma (NPC) is a common disease in Hong Kong and southern provinces of China. EBV infection is believed to play a critical role in the development of NPC. Previous studies on the transformation mechanism of EBV genes were mostly performed in either NPC or nonnasopharyngeal epithelial cells which may not be representative of premalignant nasopharyngeal epithelial cells. Establishment of a representative cell system would greatly facilitate the elucidation of the role of EBV infection in the development of NPC. Using telomerase alone, we were able to establish an immortalized nasopharyngeal epithelial cell line from primary nonmalignant nasopharyngeal biopsies. The telomerase-immortalized nasopharyngeal epithelial cells are largely diploid in karyotype. Interestingly, this newly immortalized nasopharyngeal epithelial cell line, referred as NP460hTert, harbors genetic alterations previously identified in premalignant and malignant nasopharyngeal epithelial cells. These include inactivation of p16 by homozygous deletion of the p16(INK4A) locus and downregulation of RASSF1A expression. The deletion of the p16(INK4A) locus appears to be the most crucial event for the immortalization of nasopharyngeal epithelial cells by telomerase and precedes RASSF1A downregulation. In addition, detailed analysis of the cytogenetic changes by conventional cytogenetics, spectral karyotyping (SKY) and array-based CGH revealed a gain of a 17q21-q25 fragment on 11p15 chromosome in all NP460hTert cells which occurred before deletion of the p16(INK4A) locus. Gain of 17q has been previously reported in NPC. In addition, activation of NF-kappaB was observed in immortalized NP460hTert cells at the later population doublings, and may play a role in the survival of immortalized NP epithelial cells. Id1 which is commonly expressed in various human cancers, including NPC, was also upregulated in the immortalized NP460hTert cells. Thus, the establishment of an immortalized nasopharyngeal epithelial cell line harboring common genetic alterations present in premalignant and cancerous nasopharyngeal epithelial cells may provide a valuable cell system to examine for early events involved in NPC carcinogenesis, particularly in elucidating the role of EBV infection in NPC development.

Cell Proliferation↗
PubMed2004

The homeodomain-containing transcription factor X-nkx-5.1 inhibits expression of the homeobox gene Xanf-1 during the Xenopus laevis forebrain development.

Expression of the homeobox gene Xanf-1 starts within the presumptive forebrain primordium of the Xenopus embryo at the midgastrula stage and is inhibited by the late neurula. Such stage-specific inhibition is essential for the normal development as the experimental prolongation of the Xanf-1 expression elicits severe brain abnormalities. To identify transcriptional regulators that are responsible for the Xanf-1 inhibition, we have used the yeast one-hybrid system and identified a novel Xenopus homeobox gene X-nkx-5.1 that belongs to a family of Nkx-5.1 transcription factors. In terms of gene expression, X-nkx-5.1 shares many common features with its orthologs in other species, including expression in the embryonic brain and in the ciliated cells of the otic and lateral line placodes. However, we have also observed several features specific for X-nkx-5.1, such as expression in precursors of the epidermal ciliated cells that may indicate a possible common evolutionary origin of all ciliated cells derived from the embryonic ectoderm. Another specific feature is that the X-nkx-5.1 expression in the anterior neural plate starts early, within the area overlapping the Xanf-1 expression territory at the midneurula stage, and it correlates with the beginning of the Xanf-1 inhibition. Using various loss and gain-of-function techniques, including microinjections of antisense morpholino oligonucleotides and mRNA encoding for the X-nkx-5.1 and its dominant repressor and activator versions, we have shown that X-nkx-5.1 can indeed play a role of stage-specific inhibitor of Xanf-1 in the anterior neural plate during the Xenopus development.

Amino Acid Sequence↗
PubMed1997

GDNF is a trophic factor for adult rat corticospinal neurons and promotes their long-term survival after axotomy in vivo.

Glial cell line-derived neurotrophic factor (GDNF) is a trophic factor for several neuronal populations involved in motor control. The present study evaluates the trophic actions of GDNF on corticospinal neurons, an important central nervous system motor projection into the spinal cord. Death of spinal motoneurons and corticospinal neurons is observed in the neurodegenerative disease amyotrophic lateral sclerosis. Axotomy of adult rat corticospinal neurons at internal capsule levels induces half of them to die, and the surviving population displays severe atrophy. To examine the trophic effects of GDNF on corticospinal neurons, Fast Blue-labelled corticospinal neurons were stereotaxically axotomized at internal capsule levels and GDNF was infused intracortically to lesioned corticospinal neurons at total doses of 2, 4, 10, 20, 40, 100 and 300 microg for 7 days. GDNF prevented axotomy-induced death of corticospinal neurons at doses between 2 and 40 microg and abolished or attenuated their atrophy at all doses examined. In addition, treatment with 8 microg GDNF for the first 2 weeks after axotomy resulted in the long-term survival of corticospinal neurons for 42 days. With regard to the development of treatment strategies for upper motoneuron degeneration in amyotrophic lateral sclerosis, application of GDNF via the cerebrospinal fluid may be more relevant than intracortical delivery as its diffusion within the brain parenchyma is limited. Intraventricular as well as intracisternal infusion of GDNF (300 microg over 7 days) completely prevented corticospinal neuron death. These results show that GDNF promotes the long-term survival of corticospinal neurons and has a positive effect on their size in vivo. Furthermore, the survival-promoting effect of GDNF on corticospinal neurons after delivery via cerebrospinal fluid has important clinical implications for potential treatment of the upper motoneuron degeneration seen in amyotrophic lateral sclerosis.

Animals↗
PubMed2006

Functional role of GABAergic innervation of the cochlea: phenotypic analysis of mice lacking GABA(A) receptor subunits alpha 1, alpha 2, alpha 5, alpha 6, beta 2, beta 3, or delta.

The olivocochlear efferent system is both cholinergic and GABAergic and innervates sensory cells and sensory neurons of the inner ear. Cholinergic effects on cochlear sensory cells are well characterized, both in vivo and in vitro; however, the robust GABAergic innervation is poorly understood. To explore the functional roles of GABA in the inner ear, we characterized the cochlear phenotype of seven mouse lines with targeted deletion of a GABA(A) receptor subunit (alpha1, alpha2, alpha5, alpha6, beta2, beta3, or delta). Four of the lines (alpha1, alpha2, alpha6, and delta) were normal: there was no cochlear histopathology, and cochlear responses suggested normal function of hair cells, afferent fibers, and efferent feedback. The other three lines (alpha5, beta2, and beta3) showed threshold elevations indicative of outer hair cell dysfunction. Alpha5 and beta2 lines also showed decreased effects of efferent bundle activation, associated with decreased density of efferent terminals on outer hair cells: although the onset of this degeneration was later in alpha5 (>6 weeks) than beta2 (<6 weeks), both lines shows normal efferent development (up to 3 weeks). Two lines (beta2 and beta3) showed signs of neuropathy, either decreased density of afferent innervation (beta3) or decreased neural responses without concomitant attenuation of hair cell responses (beta2). One of the lines (beta2) showed a clear sexual dimorphism in cochlear phenotype. Results suggest that the GABAergic component of the olivocochlear system contributes to the long-term maintenance of hair cells and neurons in the inner ear.

Acoustic Stimulation↗
PubMed1994

[Fine structure of conjunctival lymphatic vessel valves].

BACKGROUND: Lymphatic valves are due to ensure a zentripetal flow of lymph. The majority of the authors regard these valves as bicuspid structures with joint insertion points of the downstream localized free valvular edges. Because of that valvular structure a retrograde flow of lymph is impossible. Observations of our own however demonstrated retrograde flow of fluid under physiological conditions. In the following paper the structure of conjunctival lymphatic valves should be investigated or rather those structures, which were regarded as valves hitherto. Are there segmental connections, which allow retrograde flow of lymph? Is a regulation of lymph flow possible without self-acting valves? MATERIALS AND METHODS: In the conjunctiva of bovine post-mortem eyes (n = 100) segmental connections of lymphatic vessels with a diameter of less than 1 mm were opened specifically under control of slit lamp microscope after having undertaken an interstitial double contrast lymphography (solution of Berliner Blue/air). Some of these preparations (n = 20) could be investigated further on with the scanning electron microscope. That procedure allowed to make the patterns of flow and vessels structures in the beginnings of the lymphatic system observed with the slit lamp microscope congruent with those structures shown by the scanning electron microscope. RESULTS: The following segmental connections could be demonstrated: 1. Lymphatic vessels, which flow like a pipe into the wall of another segment. 2. Laterally arranged oval connections with valve resembling an aperture. 3. Segments of lymphatic vessels arranged in a line, which flow into one another between two hump-shaped protrusions with bicuspid valvular structures. Also the existence of different connecting structures ("valves") in one segment ("gear segment") were observed. In bulbus-like segmental connections arranged one behind the other with two valvular leaflets it could be demonstrated by scanning electron microscopy, that the two leaflets were attached apart at the vessel wall. That divergence to the previous models of lymphatic valves means, that these valvular leaflets cannot work as self-acting valves and prevent retrograde flow. CONCLUSIONS: However according to the valveless model of Libau a directed regulation of flow is possible. Perhaps the valvular structures described above play an important part by the interaction between the endothelium of lymphatic vessels and the substances transported in the lymph. The variety of segmental connections, the network of vessels with different diameters, which seem to be connected by feed-back, demand a new way of looking at the initial lymph flow.

Animals↗
PubMed2006

Differential effects of cortisol on MRC-5 fibroblasts and hypertrophic LL-29 fibroblasts.

Cortisol is a glucocorticoid secreted by the adrenal cortex that helps facilitate the body's response to stress and regulates the immune system. Glucocorticoid receptors can be found on most cell types and as a consequence, cortisol hormone plays an essential role on the body's physiologic systems. Cortisol has been shown to elicit differing responses from normal fibroblasts in comparison to hypertrophic fibroblasts. The purpose of this experiment was to analyze the differential effects of cortisol on normal MRC-5 fetal lung fibroblasts and hypertrophic LL-29 lung fibroblasts from a patient with idiopathic pulmonary fibrosis. The objectives of the experiment are to obtain and culture normal and hypertrophic lung fibroblasts, to challenge cells with subphysiological, physiological, and supraphysiological doses of cortisol (0.01 microg/dL, 0.2 microg/dL, 1 microg/dL) for 24, 48, and 72 hour incubation periods and to analyze cellular activity using the methods of cell count, protein assay, MDA, and morphological evaluation. Data collected from this study demonstrated variable response to cortisol by both cell lines. Striking results revealed that in LL-29 cells, supraphysiological dose of cortisol stimulated cell growth only in the 24-hour incubation period without showing any changes in number of micronucleoli or structural damage. In contrast, MRC-5 cells showed increased growth at a later stage (48 hours) with a dose specific increase with significantly increased micronucleoli numbers. In conclusion, the two cell lines differ in their response to cortisol concentration in a dose and time dependent manner. Cortisol concentrations did not induce structural damage throughout the experiment. These observations could help significantly in minimizing the traumatic side effects induced through stress conditions by employing intervention modalities to regulate systemic cortisol.

Cell Line↗
PubMed1983

The fine structure of the gill epithelium of a fresh-water flea, Daphnia magna (Crustacea: Phyllopoda) and changes associated with acclimation to various salinities. I. Normal fine structure.

Two kinds of epithelial cells, dark and light types, are alternately arranged in the gill of Daphnia magna. The dark cell has numerous mitochondria and an elaborate tubular system containing two kinds of cytoplasmic tubules, small about 70 nm in diameter, and large about 130 nm in diameter. The former occur in bundles and seem to be smooth-surfaced endoplasmic reticulum. The latter, lined with a ridged surface coat and frequently open at the lateral and basal cell membrane, are regarded as extensions of the cell membrane. The atypical cell membrane of the dark cell is modified by repeated subunits of a cytoplasmic coat on the inner leaflet of the unit membrane. The light cell exhibits a high degree of basal infoldings of the cell membrane, which represent a magnification of the surface area of the cell. Large mitochondria between the infoldings often come into intimate association with the infolded cell membrane to form a regular array of parallel mitochondria interposed with the double cell membranes. The results suggest that at least the dark epithelial cells play an important role in the osmoregulation of this animal.

Animals↗
PubMed1984

The vascular territory of the acromiothoracic axis.

The precise vascular territory and the variations of the acromio-thoracic axis were investigated in a series of 60 fresh cadavers and 50 formalin fixed specimens using dissection, ink injection and barium radiographic studies. The sternocostal portion and the clavicular head of the pectoralis major were found to have virtually independent vascular and nerve supplies. The pectoral artery supplied the former, whereas the deltoid artery nourished the latter. The dominant supply from the pectoral artery to the rib cage was found to enter around the fourth rib in the mid clavicular line. This supply is associated with a previously undescribed origin of the pectoralis major muscle in this region. The supply to the sternum was determined as indirect via the captured territory of the internal mammary system. The dominant supply to the skin from the pectoral artery arose laterally along the free lower border of the muscle as fasciocutaneous branches. The deltoid artery supplies the skin over the shoulder by numerous small branches which emerge from the intramuscular septa of the deltoid muscle. In addition a large axial artery was noted. In most cases this arose from the deltoid artery or its acromial branch and coursed laterally. It is noteworthy that the majority of skin paddles of the pectoralis major myocutaneous flap currently used in clinical practice are designed medially and inferiorly around the perimeter of the muscle and onto the rectus sheath. In these situations such flaps are not supplied directly by the pectoral artery. In fact, they are supplied indirectly by cutaneous branches belonging to the internal mammary/superior epigastric system which are captured by arterial connections with the pectoral artery. These occur predominantly in the pectoralis major muscle. Suggestions, based on these anatomical studies, are offered to improve the versatility and safety of flaps designed in this area.

Acromion↗
PubMed2003

The HEGP component-based clinical information system.

OBJECTIVES: The opening of the Georges Pompidou University Hospital (HEGP) in southwest Paris from the merging of three aging facilities was the opportunity to conceive and deploy a brand new clinical information system (CIS) based on a component-based approach. This paper describes the process of selection of the business components, the main functions currently in use, and the technical infrastructure that proved necessary. METHODS: The HEGP CIS features generic and healthcare-related components. The generic components include a reference manager, a security manager, a document manager, a Corba bus, and various mediation and supervision tools. The healthcare-related components include the patient, healthcare record, act management, and resource scheduler components. RESULTS: Major functions of CIS were operational at the opening of the hospital in July 2000. Two years later, the unique patient record and the provider order entry system were used in 96% of the concerned healthcare units. Sixty-five percent of the biological orders and 55% of the imaging orders were directly entered by the physicians. Access to investigation results including on-line availability of images is used by physicians in 100% of the units. DISCUSSION AND CONCLUSION: A component-based approach was found to be high-performing and cost-effective for the design and deployment of HEGP CIS.

Hospital Information Systems↗
PubMed2002

Human feeders support prolonged undifferentiated growth of human inner cell masses and embryonic stem cells.

Previous reports have demonstrated the growth of undifferentiated human embryonic stem (HES) cells on mouse embryonic fibroblast (MEF) feeders and on laminin- or Matrigel-coated plastic surfaces supplemented with MEF-conditioned medium. These xenosupport systems run the risk of cross-transfer of animal pathogens from the animal feeder, matrix, or conditioned medium to the HES cells, thus compromising later clinical application. Here we show that human fetal and adult fibroblast feeders support prolonged undifferentiated HES cell growth of existing cell lines and are superior to cell-free matrices (collagen I, human extracellular matrix, Matrigel, and laminin) supplemented with human or MEF feeder-conditioned medium. Additionally, we report the derivation and establishment of a new HES cell line in completely animal-free conditions. Like HES cells cultured on MEF feeders, the HES cells grown on human feeders had normal karyotypes, tested positive for alkaline phosphatase activity, expressed Oct-4 and cell surface markers including SSEA-3, SSEA-4, Tra 1-60, and GCTM-2, formed teratomas in severely combined immunodeficient (SCID) mice, and retained all key morphological characteristics. Human feeder#150;supported HES cells should provide a safer alternative to existing HES cell lines in therapeutic applications.

Animals↗
PubMed1993

Direct in vivo gene transfer to ependymal cells in the central nervous system using recombinant adenovirus vectors.

To evaluate the potential for adenovirus-mediated central nervous system (CNS) gene transfer, the replication deficient recombinant adenovirus vectors Ad.RSV beta gal (coding for beta-galactosidase) and Ad-alpha 1AT (coding for human alpha 1-antitrypsin) were administered to the lateral ventricle of rats. Ad.RSV beta gal transferred beta-galactosidase to ependymal cells lining the ventricles whereas Ad-alpha 1AT mediated alpha 1-antitrypsin secretion into the cerebral spinal fluid for 1 week. These observations, together with beta-galactosidase activity in the globus pallidus and substantia nigra following stereotactic administration of Ad.RSV beta gal to the globus pallidus, suggest that adenovirus vectors will be useful for CNS gene therapy.

Adenoviruses, Human↗
PubMed2002

A comparative study of effects of antiproliferative drugs on human retinal pigment epithelial cells in vitro.

The effects of various antiproliferative drugs have been tested in numerous cell types in vitro, but a comparison of effects of these drugs on cultured human retinal pigment epithelium (RPE) has not been reported. We studied the effects of four most widely used antiproliferative drugs (5-FU, daunomycin, mitomycin and dexamethasone) in a new in vitro model system (cultured human RPE). Various concentrations and exposure periods were tested in four human RPE cell lines. 5-FU showed a dose-dependant growth inhibition, with an ID50 of 5.35 +/- 2.38 x 10(-7) M after 4 days culture. A decrease of cell number occurred relatively later, the slope of the dose-response curve was less steep than that of others. Mitomycin C showed an immediate and strong growth inhibition and cytotoxic effects, with an ID50 of 3.73 +/- 0.71 X 10(-9) M. RPE cultured with daunomycin showed an abrupt decrease of cell number from 10(-9) M to 10(-8) M, the ID50 value was 1.07 +/- 0.23 x 10(-8) M. Dexamethasone showed a biphasic effect; it stimulated cell growth at 10(-7) to 10-6 M and inhibited cell growth at 10(-4) M or higher, with an ID50 of 6.05 +/- 1.61 x 10(-3) M. The advantages and disadvantages of these drugs and the prospective clinical application of these drugs for management of proliferative vitreoretinopathy (PVR) were discussed. Development of an in vitro model using cultured human RPE to study the effects of various antiproliferative drugs can provide a rapid, safe and inexpensive method for selection of drugs used for management of proliferative vitreoretinopathy.

Antineoplastic Agents↗
PubMed2005

Transduction of the choroid plexus and ependyma in neonatal mouse brain by vesicular stomatitis virus glycoprotein-pseudotyped lentivirus and adeno-associated virus type 5 vectors.

Evaluation of gene transfer into the developing mouse brain has shown that when adeno-associated virus serotype 1 (AAV1) or AAV2 vectors are injected into the cerebral lateral ventricles at birth, widespread parenchymal transduction occurs. Lentiviral vectors have not been tested by this route. In this study, we found that injection of lentiviral vectors pseudotyped with vesicular stomatitis virus glycoprotein (VSV-G) resulted in targeted transduction of the ependymal cells lining the ventricular system and the choroid plexus along the entire rostrocaudal axis of the brain, whereas a Mokola pseudotype transduced only a few cells after injection into the neonatal ventricle. In contrast, when lentiviral vectors pseudotyped with either VSV-G or Mokola glycoprotein are injected into the adult mouse brain, they transduce similar patterns of cells. An Ebola-Zaire-pseudotyped vector did not transduce any neonatal CNS cells, as was also the case for adult parenchymal injections. Long-term gene expression (12 months) occurred with a constitutively active mammalian promoter and a self-inactivating long terminal repeat (LTR), whereas the cytomegalovirus promoter in a vector with an intact LTR was expressed only in short-term experiments. We found that an AAV5 vector also targeted the ependymal and choroid plexus cells throughout the ventricular system. This vector exhibited limited penetration from the ventricle to other structures, which was significantly different from the previously reported patterns of transduction after intraventricular injection of AAV1 and AAV2 vectors.

Animals↗
PubMed2000

Characterization of huJAM: evidence for involvement in cell-cell contact and tight junction regulation.

Cell-cell interactions of the mucosal epithelia are important for the maintenance and establishment of epithelial barrier function. During events of inflammation, such cell-cell interactions are often disrupted, resulting in a leaky epithelial barrier, which in turn can lead to various inflammatory and infective dysfunctions. Human junctional adhesion molecule (huJAM), found on the mucosal epithelia and vascular endothelia of many major organ systems, is a membrane glycoprotein which resolves to a doublet band of approximately 40 and approximately 37 kDa under SDS-PAGE analysis, representing differentially glycosylated forms of the same protein. huJAM was localized to the lateral membrane of Caco-2 cells (a human colonic epithelial cell line) monolayers, in an area basolateral of the epithelial tight junctions (TJ). Through functional and biochemical assays, we show huJAM to be able to homotypically associate and to participate in TJ restitution after trypsin-EDTA disruption. Furthermore, we also observed a migration of huJAM expression toward areas of cell-cell contacts during events of cell adhesion and monolayer formation. These qualities makes huJAM a likely player in the regulation of cell-cell contacts and the subsequent formation of TJs.

Animals↗