3D Culture of Primary Patient-Derived Hepatoblastoma Tumoroids.

Hepatoblastoma, the most common primary liver malignancy in children, remains poorly understood due in part to a relative lack of methods to expand tumor cells in culture and a paucity of robust experimental models. Here, we describe a method to obtain primary tumor cells from patients with hepatoblastoma and to propagate the cells in 3D culture as tumor organoids, or "tumoroids". We further detail methods to prepare the tumoroids for whole-mount and cross-sectional imaging as well as to perform lentiviral transduction.

Current Approaches in Hepatoblastoma-New Biological Insights to Inform Therapy.

PURPOSE OF REVIEW: As the most common pediatric primary liver cancer with rising incidence, hepatoblastoma remains challenging to treat. Here, we review the current understanding of the biology of hepatoblastoma and discuss how recent advances may lead to new treatment modalities. RECENT FINDINGS: Standard chemotherapy regimens including cisplatin, in addition to surgery, have led to high cure rates among patients with low stage hepatoblastoma; however, metastatic and relapsed disease continue to have poor outcomes. Recent genomics and functional studies in cell lines and mouse models have established a central role for the Wnt/ß-catenin pathway in tumorigenesis. Targeted agents and immunotherapy approaches are emerging as potential treatment avenues. With recent gains in knowledge of the genomic and transcriptomic landscape of hepatoblastoma, new therapeutic mechanisms can now be explored to improve outcomes for metastatic and relapsed hepatoblastoma and to reduce the toxicity of current treatments.

A comparison of the cytogenetic alterations and global DNA hypomethylation induced by the benzene metabolite, hydroquinone, with those induced by melphalan and etoposide.

Specific cytogenetic alterations and changes in DNA methylation are involved in leukemogenesis. Benzene, an established human leukemogen, is known to induce cytogenetic changes through its active metabolites including hydroquinone (HQ), but the specific alterations have not been fully characterized. Global DNA hypomethylation was reported in a population exposed to benzene, but has not been confirmed in vitro. In this study, we examined cytogenetic changes in chromosomes 5, 7, 8, 11 and 21, and global DNA methylation in human TK6 lymphoblastoid cells treated with HQ for 48 h, and compared the HQ-induced alterations with those induced by two well-known leukemogens, melphalan, an alkylating agent, and etoposide, a DNA topoisomerase II inhibitor. We found that rather than inducing cytogenetic alterations distinct from those induced by melphalan and etoposide, HQ induced alterations characteristic of each agent. HQ induced global DNA hypomethylation at a level intermediate to melphalan (no effect) and etoposide (potent effect). These results suggest that HQ may act similar to an alkylating agent and also similar to a DNA topoisomerase II inhibitor in living cells, both of which may be potential mechanisms of benzene toxicity. In addition to cytogenetic changes, global DNA hypomethylation may be another mechanism underlying the leukemogenicity of benzene.

"Pinch modification" of the linear advancement flap.

Pinch modification of linear advancement flap is a hybrid of advancement and bilateral rotation flaps. This modification, which shortens and widens the flap, shows an increase in flap survival rate over the traditional advancement technique.

Making title slides: a guide for physicians.

Using modern photographic equipment, new Kodak Vericolor slide film, and a little know-how, physicians can produce their own title slides for use in lectures. Slide-making is both easy and inexpensive. A step-by-step approach is outlined herein.