Comprehensive characterization of RB1 mutant and MYCN amplified retinoblastoma cell lines.

In retinoblastoma research tumor-derived cell lines remain an important model to investigate tumorigenesis and new therapy options, due to limited tumor material and lack of adequate animal models. A panel of 10 retinoblastoma cell lines was characterized with respect to mutation, methylation and expression of RB1 and MYCN. These established retinoblastoma cell lines represent the most frequent types of RB1 inactivation and together with the MYCN amplification status, three classes can be distinguished: RB1mut/MYCNnonA, RB1mut/MYCNA and RB1wt/MYCNA. MYCN amplification was identified in five cell lines, whereby two of them, RB522 and RB3823, harbor no aberration in RB1. Targeted sequencing of 160 genes often mutated in cancer identified only few variants in tumor-associated genes other than in RB1. None of these variants was recurrent. mRNA expression analyses of retinal markers, cell cycle regulators and members of the TP53 signaling pathway revealed a high variability between cell lines but no class-specific differences. The here presented thorough validation of retinoblastoma cell lines, including microsatellite analysis for cell line authentication, provides the basis for further in vitro studies on retinoblastoma.

Complementary, alternative, integrative and dietary therapies for children with Crohn's disease - A systematic review.

Children with diagnosed inflammatory bowel diseases such as Crohn's disease are faced with the daunting prospect of living with a chronic disease. Besides psychological stress, children are suffering from therapy side-effects; in particular, corticosteroid therapies are problematic in the growth phase. This highlights that there is a need for less aggressive alternative therapies for children as well as adolescents living with such chronic conditions. Elemental diets are widely used and accepted therapy options. Several pediatric Crohn's disease patients also use complementary, alternative and integrative therapies to reduce or avoid drug therapies. To survey such therapy options and their efficiency and safety, we performed a systematic literature search and screened databases (Cochrane Library, EMBASE, OvidSP, PubMed, CAMbase, CAM-QUEST, Anthromedics) from their inception to December 2019. In total, seven of 1439 studies fulfilled search criteria. Six RCTs and one retrospective controlled trial investigating elemental diets (Flexical, Elemental 028), semi-elemental diets (Pregomin), polymeric diets (Modulen IBD), whole protein based formulas, and ω-3 fatty acid supplementation were found. Data indicated that diet therapies were equal to or more effective than corticosteroid therapies when used to treat Crohn's disease. Regrettably, we could not identify controlled studies investigating complementary, alternative and integrative medicine approaches. Our review provides an updated overview of controlled studies investigating dietary therapies used in the treatment of pediatric Crohn's disease, and demonstrates that the current study situation does not reflect the actual use of complementary, alternative and integrative therapies. Therefore, clinical trials are necessary to estimate risks and benefits of such therapies. The review indicated that enteral diets and ω-3 fatty acid supplementation may be an effective alternative to corticosteroid treatments for children with Chron's disease.

Complementary and alternative medicine in the treatment of acute bronchitis in children: A systematic review.

OBJECTIVE: Our review summarizes published literature of complementary and alternative medicine (CAM) used for the treatment of acute bronchitis in children. BACKGROUND: Acute bronchitis is one of the most frequent pediatric diseases and has high prevalence for in- and outpatient care. Acute bronchitis is mainly a viral-caused infection, but a high and inappropriate use of antibiotics has been demonstrated in many countries. As CAM therapies might reduce the use of antibiotics and can complement conventional therapies in children, they could be an appropriate treatment option. METHODS: A systematic literature search was conducted using general and complementary and alternative medicine (CAM)-specific databases. A search term including 65 CAM-associated definitions was applied. RESULTS: Literature search revealed 309 articles, whereby 18 articles hit search criteria. These clinical trials were subgrouped into the categories herbal medicine, anthroposophic medicine and homeopathy. The most often studied approaches are herbal remedies, in particular the Pelargonium sidoides extract, EPs® 7630. Its efficacy was demonstrated in three placebo-controlled trials and two observational studies. Anthroposophic approaches (mainly ribwort-containing remedies) were investigated in two controlled trials and three observational studies. Two studies were found investigating the homeopathic remedies Monapax® and Droperteel®. CONCLUSION: Study results indicate a favorable effect of investigated CAM approaches. However, only three of 18 studies were randomized controlled trials (RCTs), so a reliable statement on effectiveness was not possible and further RCTs are indispensable.

An expert consensus-based guideline for the integrative anthroposophic treatment of acute gastroenteritis in children.

OBJECTIVES: Acute gastroenteritis is one of the major causes of hospital admission in childhood. The primary objective of the treatment is rehydration, but conventional drug therapies are limited. Therefore, several pediatricians supplement conventional treatment with complementary and alternative therapies. In the two German departments for pediatric integrative medicine, children suffering from an acute gastroenteritis are treated with supportive therapy based on anthroposophic medicine. However, up to now scientifically validated guidelines for these therapies are lacking. DESIGN: We consulted an expert pool of 50 physicians with expertise in anthroposophic medicine as well as pediatrics and invited them to participate in an online-based Delphi process. Results were analyzed by means of qualitative content analysis with two independent raters using MAXQDA. Using four rounds of questioning, a consensus-based guideline was developed. RESULTS: A strong consensus (>90%) or consensus (>75-90%) was achieved for 14 of 16 subsections. The guideline describes disease characteristics, the most useful diagnostics, drug as well as non-drug treatment recommendations and advises for a good physician-patient interaction. CONCLUSION: The guideline will help clinicians, as well as family doctors, in their daily routine and make anthroposophic medicine more tangible for parents and health insurance companies.

Taraxacum officinale extract shows antitumor effects on pediatric cancer cells and enhance mistletoe therapy.

OBJECTIVES: While Taraxacum officinale (dandelion) extracts showed antitumor effects on adult cancer cells, effects on pediatric tumor cells as a single agent or in combination with mistletoe extracts are hitherto unknown. MATERIAL AND METHODS: The anti-proliferative effects of an aqueous fermented Taraxacum officinale extract (Taraxacum) on a pediatric cancer cell line panel were assessed by cell viability assays (MTT). In two neuroblastoma cell lines, SH-SY5Y and Kelly, the effects on cell cycle distribution (PI staining), mitochondrial integrity (MitoTracker staining), invasion (Boyden chamber assay) and migration (Scratch-assay) as well as the synergistic effects of the co-treatment of Taraxacum and mistletoe preparations (Iscucin® Tiliae or Iscucin® Pini) were investigated. RESULTS: All tested cancer cell lines were more susceptible to Taraxacum than the normal human fibroblast cell line, NHDF-C. In neuroblastoma cell lines Taraxacum caused apoptosis and loss of mitochondrial integrity as well as an inhibition of invasion and migration. The simultaneous therapy of Taraxacum and the mistletoe extracts revealed synergistic effects. CONCLUSION: This preclinical data support the use of Taraxacum as a potential adjuvant application in pediatric oncology.