Hydrogel Environment Supports Cell Culture Expansion of a Grade IV Astrocytoma.

Malignant astrocytomas are aggressive cancers of glial origin that can develop into invasive brain tumors. The disease has poor prognosis and high recurrence rate. Astrocytoma cell lines of human origin are an important tool in the experimental pathway from bench to bedside because they afford a convenient intermediate system for in vitro analysis of brain cancer pathogenesis and treatment options. We undertook the current study to determine whether hydrogel culture methods could be adapted to support the growth of astrocytoma cell lines, thereby facilitating a system that may be biologically more similar to in vivo tumor tissue. Our experimental protocols enabled maintenance of Grade IV astrocytoma cell lines in conventional monolayer culture and in the extracellular matrix hydrogel, Geltrex™. Light and fluorescence microscopy showed that hydrogel environments promoted cellular reorganization from dispersed cells into multilayered aggregates. Transmission electron microscopy revealed the prevalence of autophagy and nuclear membrane distortions in both culture systems. Analysis of microarray Gene Expression Omnibus (GEO) DataSets highlighted expression of genes implicated in pathways for cancer progression and autophagy. A pilot quantitative polymerase chain reaction (qPCR) analysis of the autophagic biomarkers, Beclin 1 (BECN1) and microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B), with two reference genes (beta actin, ACTB; glyceraldehyde 3-phosphate dehydrogenase, GAPDH), uncovered a relative increase of BECN1 and LC3B in hydrogel cultures of astrocytoma as compared to the monolayer. Taken together, results establish that ultrastructural and molecular characteristics of autophagy are features of this astrocytoma cell line, and that hydrogel culture systems can afford novel opportunities for in vitro studies of glioma.

Use of Oritavancin (Novel New Lipoglycopeptide) in the Treatment of Prosthetic Joint Infections (PJI): A Possible Alternative Novel Approach to a Difficult Problem.

Prosthetic joint infections (PJI) result in significant morbidity, mortality and cost to patients and the health system. Traditional treatment involves a twostaged revision and occasionally a single staged revision along with intravenous antibiotics (IV) and or oral antibiotics for several weeks to months. The use of a single staged revision along with an antibiotic which has a prolonged half life and is bactericidal would be ideal. We present 2 patients who were treated successfully with a single stage revision/antibiotic spacer and a new novel long acting lipoglycopeptide called oritavancin.

Use of Intra-Articular Amphotericin B in the Treatment of Candida parasilosis and albicans in Prosthetic Joint Infections (PJI): A Novel Approach to this Difficult Problem.

BACKGROUND: Use of intra-articular antibiotics for the treatment of arthroplasty infections has gained some interest over the last few years. OBJECTIVE: Some data exists on its use with bacterial arthroplasty infections. METHOD: We used intra-articular amphotericin B in an attempt to cure these joint infections and perform a one stage revision. RESULTS: Two patients were treated with intra-articular amphotericin B for 6 weeks followed by suppressive fluconazole for 4 months. Intra-articular joint fluid was cultured during this process for re-growth of fungus. CONCLUSION: Both patients were treated successfully with the method with follow up showing no evidence of recurrence. IA administration of amphotericin B may be an alternative treatment in these patients.