A new polymer-based approach for in vivo transfection in postnatal brain.

BACKGROUND: Gene delivery within the central nervous system at postnatal age is one of the most challenging tasks in neuroscience and currently only a few effective methods are available. COMPARISON WITH EXISTING METHODS: For postnatal central nervous system cells, viral approaches are commonly used for genetic engineering but they face several biosafety requirements for production and use making them less accessible to the community. Conversely, lipid-based methods are widely used in cell culture but face limitation in vivo mainly due to the inflammatory responses they induce. To this aspect, the use of a transgenic mouse line can represent a credible answer to the community working on rat models still requires an effective and successful solution to circumvent these difficulties. NEW METHOD: We describe a new polymer-based gene delivery system allowing persistent and robust in vivo transfection with low DNA amount, reduced inflammation and high diffusion. The expression profile along the brain, the stability, the diffusion of the DNA together with the quantity of cells transfected were evaluated through in vivo approaches. RESULTS: With a single low-volume injection, we targeted different cell types within the rat brain. We measured the diffusion rate ranging from 1 to 5 mm based on the injected volume, in the three-dimensions axis. Finally, we modified brain susceptibility to epileptic seizures using a specific knock-down of the neuronal specific potassium-chloride transporter 2. CONCLUSIONS: This safe and easy system opens perspectives for non viral gene delivery in the rat brain with perspectives to study brain function in vivo.

One-dimensional gravity in infinite point distributions.

The dynamics of infinite asymptotically uniform distributions of purely self-gravitating particles in one spatial dimension provides a simple and interesting toy model for the analogous three dimensional problem treated in cosmology. In this paper we focus on a limitation of such models as they have been treated so far in the literature: the force, as it has been specified, is well defined in infinite point distributions only if there is a centre of symmetry (i.e., the definition requires explicitly the breaking of statistical translational invariance). The problem arises because naive background subtraction (due to expansion, or by "Jeans swindle" for the static case), applied as in three dimensions, leaves an unregulated contribution to the force due to surface mass fluctuations. Following a discussion by Kiessling of the Jeans swindle in three dimensions, we show that the problem may be resolved by defining the force in infinite point distributions as the limit of an exponentially screened pair interaction. We show explicitly that this prescription gives a well defined (finite) force acting on particles in a class of perturbed infinite lattices, which are the point processes relevant to cosmological N -body simulations. For identical particles the dynamics of the simplest toy model (without expansion) is equivalent to that of an infinite set of points with inverted harmonic oscillator potentials which bounce elastically when they collide. We discuss and compare with previous results in the literature and present new results for the specific case of this simplest (static) model starting from "shuffled lattice" initial conditions. These show qualitative properties of the evolution (notably its "self-similarity") like those in the analogous simulations in three dimensions, which in turn resemble those in the expanding universe.

Reducing Staphylococcus aureus bacterial counts in a dental clinic using an Ionic Breeze air purifier: a preliminary study.

Aerosols and droplets generated by dental procedures are contaminated with blood and bacteria and represent a potential route for the transmission of disease. This study sought to determine if Ionic Breeze air purifiers are effective in collecting and destroying bacteria found in dental aerosols (such as Staphylococcus aureus). This study placed one Sharper Image Professional Series Ionic Breeze Quadra unit and one Ionic Breeze GP unit (with germicidal protection) in dental operatories within the Louisiana State University School of Dentistry. After six hours of operation, bacterial samples were collected and streaked over surfaces of petri dishes containing trypticase soy sucrose bacitracin agar that had been supplemented with 5% sheep blood. The samples were incubated at 37 degrees C for 48 hours; at that point, the microbial colonies were counted. Additional testing was performed on suspect colonies to identify S. aureus strains and to determine if any of those isolates were pathogenic with or without antibiotic resistance. The Ionic Breeze GP unit killed more than 99% of all bacteria on the stainless steel collecting blades. The non-germicidal Ionic Breeze Quadra air purifier collected numerous bacteria that were found to include some pathogenic strains of S. aureus; however, none of these were resistant to antibiotics.

Role of DHEA and growth factors in chromaffin cell proliferation.

Dehydroepiandrostreone (DHEA) is a neuroactive steroid produced by the inner layer of the adrenal cortex close to the adrenomedullary cells. Chromaffin cell growth and proliferation are under the control of insulin-like growth factor II (IGF-II) and basic fibroblast growth factor (bFGF). The aim of the present study was to examine the role of DHEA on chromaffin cell proliferation induced by IGF-II and bFGF. In our model, DHEA significantly decreased IGF-II-induced proliferation by 48.7%, whereas it did not affect the proliferation induced by bFGF. These data suggest that DHEA exerts a paracrine function in the control of chromaffin cell growth.

Molecular cloning and functional characterization of a type-I neurotensin receptor (NTR) and a novel NTR from the bullfrog brain.

Neurotensin (NT) is a tridecapeptide that functions as a neurotransmitter and neuromodulator in the nervous system. To date, three different types of NT receptor (NTR), NTR1, NTR2 and NTR3, have been identified only in mammalian species. In the present study we isolated the cDNAs for an NTR1 and a novel NTR in the bullfrog brain, designated bfNTR1 and bfNTR4 respectively. bfNTR1 and bfNTR4 encode 422- and 399-amino acid residue proteins respectively. bfNTR1 has a 64% amino acid identity with mammalian NTR1, and 34-37% identity with mammalian NTR2. bfNTR4 exhibits 43% and 45-47% identity with mammalian NTR1 and NTR2 respectively. Both receptors are mainly expressed in the brain and pituitary. bfNTR1 triggers both CRE-luc, a protein kinase A (PKA)-specific reporter, and c-fos-luc, a PKC-specific reporter, activities, indicating that bfNTR1 can activate PKA- and PKC-linked signaling pathways. However, bfNTR4 appears to be preferentially coupled to the PKA-linked pathway as it induces a higher CRE-luc activity than c-fos-luc activity. bfNTRs exhibit different pharmacological properties as compared with mammalian NTRs. Mammalian NTR1 but not NTR2 responds to NT, whereas both bfNTR1 and bfNTR4 show a high sensitivity to NT. SR 48692 and SR 142948A, antagonists for mammalian NTR1 but agonists for mammalian NTR2, function as antagonists for both bfNTR1 and bfNTR4. In conclusion, this report provides the first molecular, pharmacological and functional characterization of two NTRs in a non-mammalian vertebrate. These data should help to elucidate the phylogenetic history of the G protein-coupled NTRs in the vertebrate lineage as well as the structural features that determine their pharmacological properties.

Neuroendocrine regulation of frog adrenocortical cells by neurotensin.

We previously characterized the primary structure of neurotensin (NT) from an extract of the intestine of the frog Rana esculenta. In this study, we provide evidence for the involvement of NT in the neurocrine regulation of the secretory activity of frog adrenocortical cells. Immunohistochemical studies revealed that the adrenal gland of R. esculenta is innervated by a dense network of NT-immunoreactive fibers. Graded concentrations of frog NT induced a dose-dependent stimulation of corticosterone and aldosterone secretion by frog adrenocortical explants through activation of two receptors with pEC(50) of 9.8 and 6.9. These data support the view that NT, released by nerve fibers within the frog adrenal gland, acts locally to control corticosteroid secretion.

Immunohistochemical localization, biochemical characterization, and biological activity of neurotensin in the frog adrenal gland.

The primary structure of neurotensin has been recently determined for the frog Rana ridibunda (Endocrinology 139: 4140-4146, 1998). In the present study, we have investigated the distribution and biochemical characterization of neurotensin-like immunoreactivity in the frog adrenal gland, using an antiserum directed against the conserved C-terminal region of the peptide. Neurotensin-like immunoreactivity was detected in two populations of nerve fibers: numerous varicose fibers coursing between adrenal cells, and a few processes located in the walls of blood vessels irrigating the gland. Reversed-phase HPLC analysis of frog adrenal gland extracts revealed the existence of a major peak of neurotensin-like immunoreactivity that exhibited the same retention time as synthetic frog neurotensin. The possible involvement of neurotensin in the regulation of steroid secretion was studied in vitro using perifused frog adrenal slices. For concentrations ranging from 10(-10) to 10(-5) M, synthetic frog neurotensin increased corticosterone and aldosterone production in a dose-dependent manner (EC50 = 1.2 x 10(-9) M and 5.8 x 10(-10) M, respectively). Repeated administration of neurotensin induced a reproducible stimulation of steroid output without any tachyphylaxis. Prolonged administration (3 h) of frog neurotensin caused a transient increase in corticosterone and aldosterone secretion followed by a decline of corticosteroid secretion. Neurotensin also produced a significant stimulation of corticosteroid secretion from dispersed frog adrenal cells. This study demonstrates that neurotensin is located in nerve processes innervating the adrenal gland of amphibians. The results also show that synthetic frog neurotensin exerts a direct stimulatory effect on corticosteroid output. Taken together, these data support the view that neurotensin, released by nerve fibers, may act as a local regulator of corticosteroid secretion.

[Extracorporeal lithotripsy and prophylactic antibiotic therapy]. / Lithotripsie extracorporelle et antibiothérapie prophylactique.

During ESWL, bacteria which may be contained in the stone gets disintegrated. This study evaluates the role of prophylactic antibiotherapy in preventing such complications. Fifty patients whose urine culture was negative were randomized into two groups: the first group (25 patients) received a placebo, the second group received Ceftriaxone 1 g I.V. as a single injection performed 1 hour previous ESWL. There was no positive blood culture and no positive urine culture in the placebo group arm as well as in the antibiotic group. In this study, the risk of infection induced by ESWL seems to be minimal, so that prophylactic antibiotherapy does not appear to be necessary when urine before ESWL is sterile.