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Sterol 27-hydroxylase (CYP27A1) is involved in bile acid synthesis and cholesterol homoeostasis. Cyp27a1(−/−)/ Apolipoprotein E(−/−) double knockout mice (DKO) fed a western diet failed to develop atherosclerosis. Caveolin-1 (CAV-1), the main component of caveolae, is associated with lipid homoeostasis and has regulatory roles in vascular diseases. We hypothesized that liver CAV-1 would contribute to the athero-protective mechanism in DKO mice. Cyp27a1(+/+)/ ApoE(−/−) (ApoE KO), Cyp27a1(+/−)/ ApoE(−/−) (het), and DKO mice were fed a western diet for 2 months. Atherosclerotic plaque and CAV-1 protein were quantified in aortas. Hepatic Cav-1 mRNA was assessed using qPCR, CAV-1 protein by immunohistochemistry and western blotting. Total hepatic and plasma cholesterol was measured using chemiluminescence. Cholesterol efflux was performed in RAW264.7 cells, using mice plasma as acceptor. CAV-1 protein expression in aortas was increased in endothelial cells of DKO mice and negatively correlated with plaque surface ( P < 0.05). In the liver, both CAV-1 protein and mRNA expression doubled in DKO, compared to ApoE KO and het mice ( P < 0.001 for both) and was negatively correlated with total hepatic cholesterol ( P < 0.05). Plasma from DKO, ApoE KO and het mice had the same efflux capacity. In the absence of CYP27A1, CAV-1 overexpression might have an additional athero-protective role by partly overcoming the defect in CYP27A1-mediated cholesterol efflux. [ABSTRACT FROM AUTHOR]

Gaucher disease is caused by inherited deficiency of lysosomal glucocerebrosidase. Proteome analysis of laser-dissected splenic Gaucher cells revealed increased amounts of glycoprotein nonmetastatic melanoma protein B (gpNMB). Plasma gpNMB was also elevated, correlating with chitotriosidase and CCL18, which are established markers for human Gaucher cells. In Gaucher mice, gpNMB is also produced by Gaucher cells. Correction of glucocerebrosidase deficiency in mice by gene transfer or pharmacological substrate reduction reverses gpNMB abnormalities. In conclusion, gpNMB acts as a marker for glucosylceramide-laden macrophages in man and mouse and gpNMB should be considered as candidate biomarker for Gaucher disease in treatment monitoring. [ABSTRACT FROM AUTHOR]

The article discusses research on the effects of the infection Campylobacter rectus on the fetal and neonatal survival and brain development of mice. Topics include placental inflammation and decidual hyperplasia in fetal brains, the impact of maternal infections on mortality and changes to the hippocampal region of the brain, and the longer-term impact of material infection on perinatal neurological growth and development.

Background: Preclinical end clinical studies indicate that deproteinized cancellous bovine bone is osteoconductive and may be osteopromotive. Previous studies using commercial preparations failed to demonstrate the presence of protein, implicating bone-mineral composition and 3-dimensional structure as reasons for clinical success; however, these studies did not examine whether osteoinductive factors might be present is close association with the the mineral phase. Methods: Deproteinized cancellous bovine bone was decalcified and any protein present released by chaotropic solvents using the protocol described for purification of bone morphogenetic proteins (BMPs). Three extracts were obtained and tested for their ability to support osteoinduction in the calf muscle of nude mice. Results: Protein content averaged 11 µg/g based on absorbance at 280 nm using bovine serum albumin as a standard. All extracts contained material that stained positively with silver stain after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Western blots of these gels indicated the presence of transforming growth factor-beta (TGF-β) and BMP-2. All 3 extracts were osteoinductive in the nude mouse model when combined with inactive DFDBA, and bone formation was comparable to that induced by active DFDBA. Deproteinized cancellous bovine bone by itself was not osteoinductive in the nude mouse, but in a clinical case, exhibited osteoclastic resorption with adjacent new bone formation. Conclusions: The results suggest that small amounts of protein are present in deproteinized cancellous bovine bone in close association with the mineral phase. Some of the extracted material has osteoinductive potential and may contain growth factors. This may explain the osteopromotive ability of deproteinized cancellous bovine bone clinically. [ABSTRACT FROM AUTHOR]

COMMERCIAL PREPARATIONS OF HUMAN DEMINERALIZED freeze-dried bone allograft (DFDBA) vary in their ability to induce new bone formation. This study tested the hypothesis that inactive DFDBA can be used as an effective earner of recombinant human bone morphogenetic protein-2 (rhBMP-2). Two batches of active DFDBA were used as controls. Two batches of DFDBA, previously shown to be inactive, were treated with vehicle or with 5 or 20 µg rhBMP-2 and implanted into the calf muscle of male Nu/Nu (nude) mice. Each mouse received one implant in each hind limb, both of which were of the same formulation, resulting in 8 groups of 4 mice per group: active DFDBA batch A, active DFDBA batch B, inactive DFDBA batch A, inactive DFDBA batch B, inactive DFDBA batch A plus 5 µg rhBMP-2, inactive DFDBA batch A plus 20 µg rhBMP-2, inactive DFDBA batch B plus 5 µg rhBMP-2, and inactive DFDBA batch B plus 20 µg rhBMP-2. After 56 days, the implants were removed and historically examined. A semiquantitative bone induction index was calculated based on the amount of new bone covering each historical section. Histomorphometry was also used to evaluate the area of new bone formed and the area of residual implant material. The results showed that active DFDBA induces new bone formation, whereas inactive DFDBA does not. Addition of rhBMP-2 to inactive DFDBA results in new bone formation with a bone induction index comparable to that of active DFDBA. Histomorphometric analysis, however, revealed that the rhBMP-2-containing implants caused a dose-dependent increase in new bone area that exceeded that induced by active DFDBA. At the highest concentration of rhBMP-2, bone formation was exuberant, rhBMP-2 also caused the resorption of residual implant material to levels comparable to that seen in sites treated with active DFDBA, suggesting that this growth factor may regulate resorptive cells either directly or indirectly. This study shows that addition of rhBMP-2 to inactive DFDBA provides reproducible, consistent bone induction, and suggests that inactive commercial preparations may contain inadequate amounts of BMP to cause bone induction compared to active preparations. [ABSTRACT FROM AUTHOR]

Summary. For several years, coagulation has been implicated in the pathogenesis of sepsis. However, results from clinical trials with natural anticoagulants, as well as studies with knock-out mice for specific coagulation factors yielded conflicting results on the role of coagulation in the pathogenesis of sepsis. The aim of this study was to evaluate the impact of severe The factor VIII:C (FVIII:C) and factor IX:C (FIX:C) deficiency on a lipopolysaccharide (LPS)-induced murine model of sepsis. FVIII:C and FIX:C deficient mice, and their haemostatic normal littermate controls were challenged with LPS, and several parameters of the host response were evaluated: seven-day survival experiments were performed using two dose levels of LPS; biochemical and histological markers of tissue damage, coagulation parameters, and pro-inflammatory cytokines were evaluated at baseline and after 3 h and 6 h after an injection of LPS. Severe FVIII and FIX deficiency were compatible with normal survival in experimental sepsis. In addition, LPS-induced tissue damage and coagulation activation were similar in FVIII or FIX deficient mice compared to their respective controls. A lower release of pro-inflammatory cytokines was observed in FIX but not in FVIII deficient mice. Severe FIX or FVIII deficiency does not protect mice from mortality or from tissue damage in the endotoxemia model, supporting the hypothesis that FVIII and FIX are not critical to the pathogenesis of experimental sepsis. [ABSTRACT FROM AUTHOR]

Background and Purpose: Phosphoinositide 3-kinase-γ (PI3Kγ) is implicated in many pathophysiological conditions, and recent evidence has suggested its involvement in colitis. In the present study, we investigated the effects of AS605240, a relatively selective PI3Kγ inhibitor, in experimental colitis and its underlying mechanisms.Experimental Approach: Acute colitis was induced in mice by treatment with trinitrobenzene sulphonic acid (TNBS), and the effect of AS605240 on colonic injury was assessed. Pro-inflammatory mediators and cytokines were measured by immunohistochemistry, elisa, real time-polymerase chain reaction and flow cytometry.Key Results: Oral administration of AS605240 significantly attenuated TNBS-induced acute colitis and diminished the expression of matrix metalloproteinase-9 and vascular endothelial growth factor. The colonic levels and expression of IL-1β, CXCL-1/KC, MIP-2 and TNF-α were also reduced following therapeutic treatment with AS605240. Moreover, AS605240 reduced MIP-2 levels in a culture of neutrophils stimulated with lipopolysaccharide. The mechanisms underlying these actions of AS605240 are related to nuclear factor-κ (NF-κB) inhibition. Importantly, the PI3Kγ inhibitor also up-regulated IL-10, CD25 and FoxP3 expression. In addition, a significant increase in CD25 and FoxP3 expression was found in isolated lamina propria CD4+ T cells of AS605240-treated mice. The effect of AS605240 on Treg induction was further confirmed by showing that concomitant in vivo blockade of IL-10R significantly attenuated its therapeutic activity.Conclusions and Implications: These results suggest that AS605240 protects mice against TNBS-induced colitis by inhibiting multiple inflammatory components through the NF-κB pathway while simultaneously inducing an increase in the functional activity of CD4+CD25+ Treg. Thus, AS605240 may offer a promising new therapeutic strategy for the treatment of inflammatory bowel diseases. [ABSTRACT FROM AUTHOR]

Abstract: Lung cancer is a devastating disease and a major therapeutic burden with poor survival rates. The discovery of rare cells with stem cell-like properties in solid tumours is emerging as an important area of cancer research and may help explain the resistance of these tumours to current therapeutics. Despite rapid developments in cancer stem cell research in other solid tumours, progress in the lung has been hampered by an incomplete understanding of the epithelial stem cell hierarchy, the heterogeneity of disease and the lack of a suitable in vivo transplantation model to assess stem cell behaviour. In this review we critically discuss what is currently known about the role of normal stem cells and cancer-initiating cells in lung tumour development, and briefly discuss strategies aimed at advancing the field of lung stem cell biology, with an emphasis on the design and manipulation of state-of-art mouse models. [ABSTRACT FROM AUTHOR]

Abstract: AZD0530, an orally available Src inhibitor, demonstrated potent antimigratory and anti-invasive effects in vitro, and inhibited metastasis in a murine model of bladder cancer. Antiproliferative activity of AZD0530 in vitro varied between cell lines (IC50 0.2 –>10μM). AZD0530 inhibited tumor growth in 4/10 xenograft models tested and dynamically inhibited in vivo phosphorylation of Src substrates paxillin and FAK in both growth-inhibition-resistant and -sensitive xenografts. The activity of AZD0530 in NBT-II bladder cancer cells in vitro was consistent with inhibition of cell migration and stabilization of cell–cell adhesion. These data suggest a dominant anti-invasive pharmacology for AZD0530 that may limit tumor progression in a range of cancers. AZD0530 is currently in Phase II clinical trials. [Copyright &y& Elsevier]

Background and Purpose: Ever since the discovery of photodynamic therapy, there has been a continuous search for more potent photosensitizers. Towards that end, we have synthesized a number of novel phthalocyanine derivatives. The unsymmetrical bisamino silicon(IV) phthalocyanine BAM-SiPc is one of the most potent compounds. In in vitro cell culture, it exhibits high phototoxicity against a number of cancer cell lines.Experimental Approach: In the present investigation, the in vivo effect of BAM-SiPc was studied in the tumour-bearing nude mice model. The biodistribution of BAM-SiPc was followed to evaluate its tumour selectivity and rate of clearance. The tumour volume in the hepatocarcinoma HepG2- and the colorectal adenocarcinoma HT29-bearing nude mice was measured after photodynamic therapy. The level of intrinsic toxicity induced was also investigated. Finally, the metabolism of BAM-SiPc in the 'normal' WRL68 liver cells and the hepatocarcinoma HepG2 cells was compared.Key Results: The results not only showed significant tumour regression of HepG2 and growth inhibition of HT29 in the tumour-bearing nude mice, but also no apparent hepatic or cardiac injury with the protocol used. Histological analyses showed that apoptosis was induced in the solid tumour. BAM-SiPc could be metabolized by WRL68 liver cells but not by the hepatocarcinoma HepG2 cells. Unfortunately, BAM-SiPc did not show any specific targeting towards the tumour tissue.Conclusions and Implications: The efficiency of BAM-SiPc in inhibiting tumour growth makes it a good candidate for further evaluation. Enhancement of its uptake in tumour tissue by conjugation with biomolecules is currently under investigation. [ABSTRACT FROM AUTHOR]

Background: Immunosuppressive agents may induce severe changes on bone metabolism and may impair the osseointegration process during the implant healing. No data are available concerning the influence of cyclosporin A on dental implants previously integrated to the bone. The aim of this study was to evaluate the influence of cyclosporin A administration on the mechanical retention of bone previously integrated to dental implants. Methods: Eighteen female New Zealand rabbits were submitted to an implant surgery. Each animal received one commercial dental implant of 10 x 3.75 mm. After 12 weeks of an undisturbed healing period, six animals were randomly sacrificed and the removal torque test was performed (group A). In addition, six animals were submitted to a daily injection of cyclosporin A in a dosage of 10 mg/kg (group C), and six animals received saline solution as a control (group B). After 12 weeks of cyclosporin A administration, groups B and C were sacrificed and submitted to a removal torque test in which higher values can be interpreted as higher mechanical bone retention to the implant surface or higher osseointegration. Results: The removal torque results were 305 (±9.8) Ncm for group A, 50.17 (±17.5) Ncm for group B, and 26 (±7.8) Ncm for group C. The statistical analysis showed significant differences between groups A and B (P<0.05) and groups B and C (P<0.01). Conclusion: Cyclosporin A administration may impair the mechanical retention of dental implants previously integrated to the bone. [ABSTRACT FROM AUTHOR]

Background: Bone graft materials are needed in periodontics that are osteoinductive, have good handling characteristics, and have physical properties that provide appropriate stiffness for the treatment site. Demineralized freeze-dried bone allograft (DFDBA), also called demineralized bone matrix (DBM), is osteoinductive but requires a carrier to meet the other clinical objectives, thereby decreasing the DBM content per volume of the bone graft material. The present study determined whether the DBM content of a carrier formulation is an important variable with respect to its effectiveness as an osteoinductive material. Methods: The immunocompromised Nu/Nu mouse-muscle implantation assay of osteoinductivity was used to test human DBM formulated with hyaluronic acid (HY) and cancellous and cortical bone granules from the same donor: DBM alone (11 mg); DBM (11 mg):HY, 55:45, weight/weight (wt/wt); DBM (6.4 mg):HY, 32:68, wt/wt; DBM mixed with cortical and cancellous bone chips 1:4 (DBMC) (11 mg total, of which 2.2 mg was DBM); DBMC (11 mg):HY, 55:45, wt/wt; heat-treated DBM (11 mg); HY alone; and positive-control DBM (11 mg). Osteoinduction was scored using a qualitative scale and by histomorphometry. Results: Results showed that all DBM was osteoinductive and the addition of HY did not change this as long as the amount of DBM used was held constant. The reduction in the absolute amount of DBM resulted in a reduced osteoinduction score, reduced ossicle area, and reduced new bone formation. The addition of HY also caused a decrease in the amount of residual non-vital bone particles, particularly when DBMC was implanted. Results were donor dependent. Conclusion: This study showed the importance of DBM content and donor variability in osteoinductivity of DBM formulations with improved handling and stiffness characteristics. [ABSTRACT FROM AUTHOR]

Early studies have demonstrated that implantation of laboratory preparations of demineralized freeze dried bone (DFDB) into the thigh muscle of mice induces ectopic osteoinduction. However, with the development of commercial preparations of DFDB for clinical use, concerns have been raised as to the osteoinductive properties of such preparations. The aim of this study was to investigate the osteoinductive potential of some commercial preparations of DFDB compared to a newly developed product which incorporates DFDB into a collagen sponge. Commercial preparations of DFDB or the DFDB/collagen sponge were inserted into the thigh muscles of 60 adult Swiss CD-1 mice. At the completion of each experimental period (7, 14, 30, 90 and 180 days), the animals were sacrificed, and the hindquarters of the mice were radiographed. The area where each graft had been placed was then excised, processed for light microscopy, and stained with hematoxylin and eosin or von Kossa's stain. Histological analysis of the DFDB/collagen sponges demonstrated significant remineralization which increased with time. Remineralization of the DFDB/collagen sponges was verified by radiographs which showed a significant increase in radiopacity over time. There was no radiographic evidence of mineralized tissue formation or remineralization in any of the commercial DFDB samples studied. At all time points studied, histological analyses failed to show evidence of bone formation for any of the preparations. The results suggest that commercially available DFDB is not osteoinductive in the murine model and question the use of such materials in clinical periodontics. The results found for the DFDB/collagen sponge indicate a different mechanism of activity from DFDB as evidenced by its rapid remineralization. The role this remineralization process has in osteoinduction is unknown and requires further study. [ABSTRACT FROM AUTHOR]