The effects of G-CSF and naproxen sodium on the serum TGF-beta1 level and fracture healing in rat tibias

Local and systemic release of transforming growth factor beta 1 (TGF-beta1) is known to increase during the process of fracture healing and this cytokine stimulates bone healing. The majority of the non steroidal anti inflammatory drugs (NSAIDs) inhibit fracture healing. Granulocyte colony stimulating factor (G-CSF) is a hematopoietic growth factor that stimulates bone marrow. In this study, the effects of the NSAID naproxen sodium, G-CSF, and both of them in combination on the TGF-beta1 serum level in rats with tibia fractures were measured and fracture healing was evaluated by histopathologic and radiologic examination. The TGF-beta1 serum levels obtained on day one (24 h after fracture but before administration of naproxen or G-CSF) were found to be similar in all of the five groups (p > 0.05). At the end of the first week, TGF-beta1 levels were significantly lower in naproxen-treated rats than those of the other groups excluding control (p = 0.002). Similar changes in TGF-beta1 levels were found at the end of the second and fourth weeks. TGF-beta1 levels were significantly higher in G-CSF-treated rats at the end of the first, second and fourth weeks (p < 0.05). Fracture healing scores measured with histopathological and radiological methods were higher in G-CSF-treated rats than in naproxen-treated ones. When both naproxen and G-CSF were given, the scores resumed to normal. The results point to the negative effect of naproxen sodium on fracture healing is due to its decreasing effect on the level of TGF-beta1, which may be a new possible mechanism. Moreover, this negative effect can be inhibited by the use of G-CSF.     

Phenotypical and functional characterization of the CD8+ T cell repertoire of HLA-A2.1 transgenic, H-2KbnullDbnull double knockout mice.

Homozygous HLA-A2.1 transgenic H-2KbnullDbnull double knockout (KO) mice were created. Their potential to develop HLA-A2. 1-restricted cytolytic responses was compared with that of their classical transgenic counterparts, which still express H-2Kb, Db molecules. On cell surfaces, both strains express similar amounts of chimeric (alpha 1 alpha 2 domains of human, alpha 3 cytoplasmic domains of mouse) HLA-A2.1 molecules in noncovalent association with mouse beta 2-microglobulin. Compared with mice that are totally deprived of histocompatibility class Ia molecules (H-2KbnullDbnull double KO), the expression of HLA-A2.1 in transgenic/double KO mice resulted in sizeable increase in the periphery of CD8+ T cells with a normally diversified TCR repertoire. A biased education in favor of HLA-A2.1, ascribable to the absence of H-2 class Ia molecules, was evidenced in these transgenic/double KO mice by their improved capacity to mount HLA-restricted cytolytic responses, regardless of whether they were virally infected or injected with synthetic epitopic peptide. HLA class I transgenic, H-2 class Ia KO mice should represent useful animal models for the preclinical evaluation of vaccine formulations aiming at the induction of HLA class I-restricted CTL responses.     

Growth and Senescence of the Successive Grass Leaves on a Tiller. Ontogenic Development and Effect of Temperature

The tiller characteristics (length and age of laminae, numberof leaves per tiller) which depend on morphogenetic characterssuch as leaf appearance and expansion rates, leaf growth durationand leaf lifespan were studied in the field over four growingseasons to gain a better understanding of the progressive changesin leaf digestibility over time, and to facilitate the developmentof predictive mathematical models. We show that, for a givenregrowth, only the number of leaves per tiller and the laminaexpansion rate remain constant throughout growth. In other words,the length of successive laminae, their growth duration andlifespan increased while their rate of appearance decreasedin such a way that the lamina expansion rate at the tiller levelremained constant. These changes were associated with an increasein sheath length which governs both the lamina appearance rateand its growth duration. As temperature increased, the averagelamina expansion rate and the number of laminae which grew bothincreased simultaneously. Therefore, high temperature acceleratesthe changes in tiller characteristics which occur as growthprogresses. Copyright 2000 Annals of Botany Company Leaf, senescence, phyllochron, lifespan, digestibility, temperature, cocksfoot, Dactylis glomerata L     

Early osteological development of the fins in the hatchery-reared red porgy, Pagrus pagrus (L. 1758)

The present study was undertaken to establish the normal, healthy features of morphological structures at various developmental stages as achieved under well-defined environmental culture conditions (temperature between 16 and 21°C, salinity 36 ppt, pH around 7.6, and oxygen saturation over 95%) common in aquaculture of the species. The pectoral fin supports began to develop at 2.90 mm total length (TL), followed by those of dorsal fins at 5.5 mm TL, caudal fins at 5.6 mm TL, pelvic fins at 5.9 mm TL and anal fins at 6.0 mm TL. The pelvic fins appeared fully at 7.4 mm TL. Development of dorsal lepidotrichia was first observed at 6.9 mm TL, attaining their final number at 7.6 mm TL. The dorsal spines first appeared at 6.5 mm TL and were complete at 7.4 mm TL. The anal lepidotrichia appeared during the development phase from 6.8 to 8.6 mm TL. At 5.6 mm TL, the upward flexion of the urostyle was initiated. The caudal lepidotrichia formed within the primordial fin at 5.6 mm TL and reached the final count at 7.4 mm TL. The caudal dermatotrichia first appeared at 7.3 mm TL and all forms were observed by 15.5 mm TL. The development pattern of fin supports found in Pagrus pagrus is quite similar to that described for other Sparid species.     

Non-invasive in vivo imaging of tumor-associated CD133/prominin

Background

Cancer stem cells are thought to play a pivotal role in tumor maintenance, metastasis, tumor therapy resistance and relapse. Hence, the development of methods for non-invasive in vivo detection of cancer stem cells is of great importance.

Methodology/Principal Findings

Here, we describe successful in vivo detection of CD133/prominin, a cancer stem cell surface marker for a variety of tumor entities. The CD133-specific monoclonal antibody AC133.1 was used for quantitative fluorescence-based optical imaging of mouse xenograft models based on isogenic pairs of CD133 positive and negative cell lines. A first set consisted of wild-type U251 glioblastoma cells, which do not express CD133, and lentivirally transduced CD133-overexpressing U251 cells. A second set made use of HCT116 colon carcinoma cells, which uniformly express CD133 at levels comparable to primary glioblastoma stem cells, and a CD133-negative HCT116 derivative. Not surprisingly, visualization and quantification of CD133 in overexpressing U251 xenografts was successful; more importantly, however, significant differences were also found in matched HCT116 xenograft pairs, despite the lower CD133 expression levels. The binding of i.v.-injected AC133.1 antibodies to CD133 positive, but not negative, tumor cells isolated from xenografts was confirmed by flow cytometry.

Conclusions/Significance

Taken together, our results show that non-invasive antibody-based in vivo imaging of tumor-associated CD133 is feasible and that CD133 antibody-based tumor targeting is efficient. This should facilitate developing clinically applicable cancer stem cell imaging methods and CD133 antibody-based therapeutics.