Purpose: High-grade osteosarcoma is a malignant primary bone tumor with a peak incidence in adolescence. Overall survival (OS) of patients with resectable metastatic disease is approximately 20%. The exact mechanisms of development of metastases in osteosarcoma remain unclear. Most studies focus on tumor cells, but it is increasingly evident that stroma plays an important role in tumorigenesis and metastasis. We investigated the development of metastasis by studying tumor cells and their stromal context.Experimental Design: To identify gene signatures playing a role in metastasis, we carried out genomewide gene expression profiling on prechemotherapy biopsies of patients who did (n ¼ 34) and patients who did not (n ¼ 19) develop metastases within 5 years. Immunohistochemistry (IHC) was performed on pretreatment biopsies from 2 additional cohorts (n ¼ 63 and n ¼ 16) and corresponding postchemotherapy resections and metastases.Results: A total of 118/132 differentially expressed genes were upregulated in patients without metastases. Remarkably, almost half of these upregulated genes had immunological functions, particularly related to macrophages. Macrophage-associated genes were expressed by infiltrating cells and not by osteosarcoma cells. Tumor-associated macrophages (TAM) were quantified with IHC and associated with significantly better overall survival (OS) in the additional patient cohorts. Osteosarcoma samples contained both M1-(CD14/HLA-DRa positive) and M2-type TAMs (CD14/CD163 positive and association with angiogenesis).Conclusions: In contrast to most other tumor types, TAMs are associated with reduced metastasis and improved survival in high-grade osteosarcoma. This study provides a biological rationale for the adjuvant treatment of high-grade osteosarcoma patients with macrophage activating agents, such as muramyl tripeptide.

The unique biology and medical relevance of the mitochondrion of the malaria parasite Plasmodium falciparum have made it the subject of many studies. However, we actually do not have a comprehensive assessment of which proteins reside in this organelle.

Chondromyxoid fibroma (CMF) is an uncommon benign cartilaginous tumor of bone usually occurring during the second decade of life. CMF is associated with recurrent rearrangements of chromosome bands 6p23-25, 6q12-15, and 6q23-27. To delineate further the role and frequency of the involvement of three candidate regions (6q13, 6q23.3 and 6q24) in the pathogenesis of CMF, we studied a group of 43 cases using a molecular cytogenetic approach. Fluorescence in situ hybridization with probe sets bracketing the putative breakpoint regions was performed in 30 cases. The expression level of nearby candidate genes was studied by immunohistochemistry and quantitative RT-PCR in 24 and 23 cases, respectively. Whole-genome copy number screening was performed by array comparative genomic hybridization in 16 cases. Balanced and unbalanced rearrangements of 6q13 and 6q23.3 occurred in six and five cases, respectively, and a hemizygous deletion in 6q24 was found in five cases. Two known tumor suppressor genes map to the latter region: PLAGL1 and UTRN. However, neither of these two genes nor BCLAF1 and COL12A1, respectively located in 6q23.3 and 6q13, showed altered expression. Therefore, although rearrangements of chromosomal regions 6q13, 6q23.3, and 6q24 are common in CMF, the complexity of the changes precludes the use of a single fluorescence in situ hybridization probe set as an adjunct diagnostic tool. These data indicate that the genetic alterations in CMF are heterogeneous and are likely a result of a cryptic rearrangement beyond the resolution level of combined binary ratio fluorescence in situ hybridization or a point mutation.

Background: Congenital isolated growth hormone deficiency (IGHD) is a rare endocrine disorder that presents with severe proportionate growth failure. Dominant (type II) IGHD is usually caused by heterozygous mutations of GH1. The presentation of newly affected family members in 3 families with dominant IGHD in whom previous genetic testing had not demonstrated a GH1 mutation or had not been performed, prompted us to identify the underlying genetic cause. Methods:GH1 was sequenced in 3 Caucasian families with a clinical autosomal dominant IGHD. Results: All affected family members had severe growth hormone (GH) deficiency that became apparent in the first 2 years of life. GH treatment led to a marked increase in height SDS. So far, no other pituitary dysfunctions have become apparent. In the first family a novel splice site mutation in GH1 was identified (c.172-1G>C, IVS2-1G>C). In two other families a previously reported splice site mutation (c.291+1G>A, IVS3+1G>A) was found. Conclusion: These data show that several years after negative genetic testing it was now possible to make a genetic diagnosis in these families with a well-defined, clearly heritable, autosomal dominant IGHD. This underscores the importance of clinical and genetic follow-up in a multidisciplinary setting. It also shows that even without a positive family history, genetic testing should be considered if the phenotype is strongly suggestive for a genetic syndrome. Identification of pathogenic mutations, like these GH1 mutations, has important clinical implications for the surveillance and genetic counseling of patients and expands our knowledge on the genotype-phenotype correlation.

and testicular atrophy with aspermatogenia (negative OCT3/OCT4 stain). There was evidence of acute multifocal bronchopneumonia and congestive heart failure. He carried two heterozygous mutations in ALMS1: 11316_11319delAGAG; R3772fs in exon 16 and 8164C>T ter; R2722X in exon 10. Conclusions: This report describes previously undefined cardiac abnormalities in this rare multisystem disorder. Myofibrillar disarray is probably directly linked to ALMS1 mutation, while fibrosis in multiple organs may be a secondary phenomenon to gene alteration. Whether and how intracellular trafficking or related signals lead to cardiac dysfunction is a subject for further research.

of the placenta provided confirmatory evidence concerning the COD; provided new information and COD; or did not provide additional information regarding the COD. Results: During the 21-year period examined, 458 stillborn autopsies were performed; 388 of these autopsies revealed structurally normal fetuses (84.7%). Of the structurally normal fetal autopsies, 94 cases (24%) were performed prior to the policy institution and 294 after. Comparing the frequency of placental examination and COD determination before and after the policy (1992), we demonstrated that the percentage of placental examinations increased from 42.2±5.3 % (1987-1991) to 92.1±2.1 % (1992-2007), and the frequency of COD determination increased from 38.5±7 % to 79.5±3.2 % (P=<0.0001). The placental examination provided confirmatory evidence in 24.6 %, new diagnostic information and COD in 44.7 % and no additional information in 30.7 %. However, in those that did not yield any additional information, COD was identified from the clinical history in 27 % and in the remainder, several common causes of intrauterine demise, such as intrauterine infection or villitis of unknown etiology, were excluded. Conclusions: The cause of death in structurally normal intrauterine fetal demise cases is more likely to be identified if routine pathologic examination of the placenta is performed.