Goat uromodulin promoter directs kidney-specific expression of GFP gene in transgenic mice.

BACKGROUND: Uromodulin is the most abundant protein found in the urine of mammals. In an effort to utilize the uromodulin promoter in order to target recombinant proteins in the urine of transgenic animals we have cloned a goat uromodulin gene promoter fragment (GUM promoter) and used it to drive expression of GFP in the kidney of transgenic mice. RESULTS: The GUM-GFP cassette was constructed and transgenic mice were generated in order to study the promoter's tissue specificity, the GFP kidney specific expression and its subcellular distribution. Tissues collected from three GUM-GFP transgenic mouse lines, and analyzed for the presence of GFP by Western blotting and fluorescence confirmed that the GUM promoter drove expression of GFP specifically in the kidney. More specifically, by using immuno-histochemistry analysis of kidney sections, we demonstrated that GFP expression was co-localized, with endogenous uromodulin protein, in the epithelial cells of the thick ascending limbs (TAL) of Henle's loop and the early distal convoluted tubule in the kidney. CONCLUSION: The goat uromodulin promoter is capable of driving recombinant protein expression in the kidney of transgenic mice. The goat promoter fragment cloned may be a useful tool in targeting proteins or oncogenes in the kidney of mammals.

Reduction of inflammation and preservation of neurological function by anti-CD52 therapy in murine experimental autoimmune encephalomyelitis.

Alemtuzumab, a monoclonal antibody directed against human CD52, is used in the treatment of MS. To characterize the impact of anti-CD52 administration, a monoclonal antibody to mouse CD52 (anti-muCD52) was generated and evaluated in EAE mouse models of MS. A single course of anti-muCD52 provided a therapeutic benefit accompanied by a reduction in the frequency of autoreactive T lymphocytes and production of pro-inflammatory cytokines. Examination of the CNS revealed a decrease in infiltrating lymphocytes, demyelination and axonal loss. Electrophysiological assessment showed preservation of axonal conductance in the spinal cord. These findings suggest that anti-CD52 therapy may help preserve CNS integrity.

Immune status following alemtuzumab treatment in human CD52 transgenic mice.

Alemtuzumab is a monoclonal antibody against the CD52 antigen present at high levels on the surface of lymphocytes. While treatment of multiple sclerosis patients with alemtuzumab results in marked depletion of lymphocytes from the circulation, it has not been associated with a high incidence of serious infections. In a human CD52 transgenic mouse, alemtuzumab treatment showed minimal impact on the number and function of innate immune cells. A transient decrease in primary adaptive immune responses was observed post-alemtuzumab but there was little effect on memory responses. These results potentially help explain the level of immunocompetence observed in alemtuzumab-treated MS patients.

[Orthodontic tooth displacement: histology, biology and iatrogenic effects]. / Le déplacement dentaire orthodontique : histologie, biologie et effets iatrogènes.

Iatrogenic effects of orthodontic forces, in particular root resorption, are described in the literature. However, our knowledge on the risk of osseous defects induced by orthodontic treatment is still scarce. After an overview of histological and biological principles of orthodontically-induced tooth movement, this study focuses on the recent data concerning the occurrence and the treatment of bone resorption associated with orthodontic treatment.

[The use of bone distraction in the treatment of the transverse direction in adults]. / Apport de la distraction osseuse dans le traitement du sens transversal chez l'adulte.

Crowding and mal-positioning of teeth in the dental arch are problems frequently encountered with adult patients. These discrepancies are associated with a lack of development of basal bone, a shortcoming whose etiology is difficult to discern. Treating these problems with expansion assisted by surgery depends on osseous distraction, the separating of segments of bone to create new bone and the movement of whole groups of teeth and their periodontium, a technique that assures excellent stability. It is important to define precisely the indications for this technique as well as our therapeutic protocols, both in the maxilla and in the mandible. For an expander we use a single thread expansion screw welded to two molar bands. To be sure it works effectively; the surgeon first opens the maxillary suture to liberate the two portions of the maxilla. After a ten day wait, the expansion screw is opened daily for up to two weeks depending on how much expansion is required. When the desired point is attained, the expansion device is locked, and the patient returns for check-up visits every ten days. During this period the patient is outfitted with a false tooth to mask the large and unsightly diastema that has appeared between the central incisors. Three months after the original intervention the patient receives a full-banded strap-up. The orthodontist usually employs centripetal forces to level the teeth and to correct rotations in an effort to minimize the risk of periodontal complications. The results that we have obtained with this treatment have fulfilled our expectations with significant improvement in arch form, full respect for the periodontium, an improvement in facial balance, and the appearance of a generous and pleasing smile.

Spider silk fibers spun from soluble recombinant silk produced in mammalian cells.

Spider silks are protein-based "biopolymer" filaments or threads secreted by specialized epithelial cells as concentrated soluble precursors of highly repetitive primary sequences. Spider dragline silk is a flexible, lightweight fiber of extraordinary strength and toughness comparable to that of synthetic high-performance fibers. We sought to "biomimic" the process of spider silk production by expressing in mammalian cells the dragline silk genes (ADF-3/MaSpII and MaSpI) of two spider species. We produced soluble recombinant (rc)-dragline silk proteins with molecular masses of 60 to 140 kilodaltons. We demonstrated the wet spinning of silk monofilaments spun from a concentrated aqueous solution of soluble rc-spider silk protein (ADF-3; 60 kilodaltons) under modest shear and coagulation conditions. The spun fibers were water insoluble with a fine diameter (10 to 40 micrometers) and exhibited toughness and modulus values comparable to those of native dragline silks but with lower tenacity. Dope solutions with rc-silk protein concentrations >20% and postspinning draw were necessary to achieve improved mechanical properties of the spun fibers. Fiber properties correlated with finer fiber diameter and increased birefringence.