Summary In comparison with hip fractures, increased expression of genes in the Wnt pathway and increased Wnt activity were found in bone samples and osteoblast cultures from patients with osteoarthritis, suggesting the involvement of this pathway in subchondral bone changes. No consistent differences were found in the genetic association study.

Wnt 22 β-Catenin 23 DNA methylation 24 Fractures 25 Bone diseases 26We reported previously that the expression of Wnt-related genes is lower in osteoporotic hip fractures than in 27 osteoarthritis. We aimed to confirm those results by analyzing β-catenin levels and explored potential genetic 28 and epigenetic mechanisms involved. 29 β-Catenin gene expression and nuclear levels were analyzed by real time PCR and confocal immunofluorescence. 30 Increased nuclear β-catenin was found in osteoblasts isolated from patients with osteoarthritis (99 ± 4 31 units vs. 76 ± 12, p = 0.01, n = 10), without differences in gene transcription, which is consistent with 32 a post-translational down-regulation of β-catenin and decreased Wnt pathway activity.33 Twenty four single nucleotide polymorphisms (SNPs) of genes showing differential expression between fractures 34 and osteoarthritis (WNT4, WNT10A, WNT16 and SFRP1) were analyzed in DNA isolated from blood of 853 pa-35 tients. The genotypic frequencies were similar in both groups of patients, with no significant differences. 36 Methylation of Wnt pathway genes was analyzed in bone tissue samples (15 with fractures and 15 with osteo-37 arthritis) by interrogating a CpG-based methylation array. Six genes showed significant methylation differences 38 between both groups of patients: FZD10, TBL1X, CSNK1E, WNT8A, CSNK1A1L and SFRP4. The DNA demethylating 39 agent 5-deoxycytidine up-regulated 8 genes, including FZD10, in an osteoblast-like cell line, whereas it down-40 regulated other 16 genes. 41 In conclusion, Wnt activity is reduced in patients with hip fractures, in comparison with those with osteoarthritis. 42 It does not appear to be related to differences in the allele frequencies of the Wnt genes studied. On the other 43 hand, methylation differences between both groups could contribute to explain the differences in Wnt activity.44

Objective: Genes explaining the susceptibility to osteoporosis have not been fully elucidated. Our objective was to explore the association of polymorphisms capturing common variations of the lipoprotein receptor-related protein (LRP) 5 and 6 genes, encoding two Wnt receptors, with femoral neck bone mineral density (BMD) and osteoporotic fractures of the spine and the hip. Design: Cross-sectional, case-control, and replication genetic association study. Methods: Thirty-nine tagging and functional single nucleotide polymorphisms (SNPs) were analyzed in a group of 1043 postmenopausal women and 394 women with hip fractures. The results were replicated in a different group of 342 women. Results: Three SNPs of the LRP6 gene were associated with BMD (nominal uncorrected P values !0.05) in the discovery cohort. One showed a significant association after multiple test correction; two of them were also associated in the replication cohort, with a combined standardized mean difference of 0.51 (PZ0.009) and 0.47 (P!0.003) across rs11054704 and rs2302685 genotypes. In the discovery cohort, several LRP5 SNPs were associated with vertebral fractures (odds ratio (OR) 0.67; PZ0.01), with hip fractures (unadjusted ORs between 0.59 and 1.21; PZ0.005-0.033, but not significant after multiple test adjustment or age adjustment), and with height and the projected femoral neck area, but not with BMD. Transcripts of LRP5 and LRP6 were similarly abundant in bone samples. Conclusions: In this study, we found common polymorphisms of LRP5 associated with osteoporotic fractures, and polymorphisms of the LRP6 gene associated with BMD, thus suggesting them as likely candidates to contribute to the explaination of the hereditary influence on osteoporosis.

Two cases of erythromelanosis follicularis faciei et colli in one family, affecting a brother and sister are described. In these cases erythromelanosis follicularis faciei et colli, a process of unknown aetiology, seems to have a clear hereditary component--autosomal recessive mode of inheritance--a feature which, as far as we know, has not been reported in the literature.

A clinical and histological study was made of a congenital lesion on the knee of a 3-year-old boy showing a peculiar combination of eccrine, pilar and angiomatous structures with the characteristics of organoid nevus. A discussion is made of different questions related to eccrine hemartomas with respect to the clinical picture, the association of other hamartomatous components, the alterations that these induce in other cutaneous structures and their histogenesis.

In addition to illicit methamphetamine, there are prescription and over-the-counter medications that, if ingested, may yield positive methamphetamine (MAMP) results on laboratory urine drug tests. The purpose of the study is to estimate the prevalence of medicinal and illicit MAMP in the pain population using chiral analysis to determine the relative amounts of the d and l-MAMP enantiomers. This retrospective analysis included the LC-MS/MS results and prescriber provided medication histories of 485,889 de-identified urine specimens from patients treated for pain. Two groups of 100 specimens each were subjected to chiral analysis. Group 1 contained specimens that were MAMP positive and amphetamine negative. Group 2 contained randomly selected MAMP positive specimens. The overall MAMP positivity rate of the 485,889 specimens tested was 1.6%. The prevalence of MAMP medications based on reported medications and detection of l-MAMP in Group 1 and Group 2 was 44% and 6%, respectively. These data indicate that the use of both illicit and medicinal MAMP is found in this patient population, and that medicinal use is underreported in clinical histories. Therefore, clinical laboratories should provide on request chiral analysis to aid in differentiating illicit and medicinal MAMP.

Four siblings (three males, one female), affected by harlequin foetus, are presented. Genetic aspects and classification of this variety of ichthyosis are discussed.

Osteoarthritis (OA) has a strong genetic component, and experimental evidence suggests the involvement of the Wnt pathway in its pathogenesis. Hence, we explored the association of common single nucleotide polymorphisms (SNPs) related to the Wnt pathway with hip and knee OA. Seventy-eight SNPs were analyzed in 606 patients undergoing joint replacement and in 680 control subjects. SNPs were located in WNT1, WNT10A, WNT16, DVL2, FZD5, BCL9, SFRP1, TCF7L1 and SFRP4 genes. SNPs significantly associated with OA were genotyped in an independent group of 369 patients and 407 controls. One SNP in WNT10A, rs3806557, was associated with hip OA in men (OR 0.65, 95% CI 0.46-0.93; p = 0.017), but the association was not confirmed in the replication phase. The TCF7L1 polymorphism rs11547160 was also associated with hip OA in the discovery set, but not in the replication set. Similarly, the SFRP4 SNP rs1052981 was associated with knee OA in women with OR of 2.73 (95% CI 1.29-5.8; p = 0.006), but the association was not replicated. The BCL9 polymorphism rs2353525 was associated with knee OA in women, both in the unadjusted and in the age- and BMI-adjusted analysis (OR 2.01; 95% CI 1.34-2.98; p = 0.0006). A similar, but not statistically significant, trend was observed in the replication phase. In the combined analysis, OR was 3.13 (1.34-7.28; p = 0.009). These data suggest that some SNPs of genes related to the Wnt pathway and, specifically BCL9, influence the genetic predisposition to osteoarthritis of the large joints in a sex- and joint-specific way.