beta-catenin (CTNNB1) gene amplification: a new mechanism of protein overexpression in cancer

beta-Catenin nuclear translocation is frequently observed in different types of malignancies, including gastric cancer. In gastric cancer, however, the molecular mechanisms leading to accumulation of this protein in the nucleus remain unknown. In this setting, beta-catenin (CTNNB1) mutations have been reported, but studies of mutation frequency have yielded conflicting results. Mutations or silencing of other partners of beta-catenin (i.e., APC and AXIN) are also considered rare genetic events in gastric tumorigenesis. Gene amplification is a common mechanism of activation and/or overexpression of oncogenes in gastric and other cancers. In this study, we investigated whether gene amplification is a possible mechanism of beta-catenin activation in gastric cancer by determining its presence in 49 patients with gastric cancer and two gastric-derived cell lines (KATO III and ST2957). Using fluorescence in situ hybridization, we identified beta-catenin amplification in one of the tumor samples as well as in KATO III cells. beta-Catenin immunostaining revealed nuclear translocation of the protein in both cases. In the KATO III cells, beta-catenin overexpression was confirmed by quantitative real-time PCR and Western blot analyses and beta-catenin gene amplification by Southern blot analysis and multiplex ligation probe amplification. In the KATO III cell line, no correlation was found between beta-catenin nuclear translocation and increased expression of the WNT1 target gene CCND1 (cyclin D1). Our data suggest that gene amplification is a possible mechanism of beta-catenin overexpression in cancer.     

Treatment of Pyoderma gangrenosum with low-dose colchicine

Pyoderma gangrenosum (PG) is a neutrophilic dermatosis of unknown origin. Systemic agents occasionally administered provide either incomplete long-term control of the disease or have been associated with serious adverse side effects after chronic administration. We present two patients with PG successfully treated with low-dose colchicine. Antimitotic, anti-inflammatory and immunomodulating properties of colchicine might account for its beneficial effects in PG patients. Colchicine is effective and well tolerated in low doses by most patients. In addition, it is inexpensive and safer for long-term treatment than corticosteroids and other immunosuppressive agents. Colchicine may be proposed either as a single agent or as a corticosteroid-sparing agent for early treatment of PG.     

Diagnostic difficulties in a case of isolated sarcoidosis of the nose and sinuses

Sarcoidosis still remains a diagnosis of exclusion. It is unusual for the disease to be localised in the nose and sinuses and to manifest its symptoms in this site; the diagnosis in such cases is rather a difficult task. There is no symptom pathognomonic of the disease. A major role in the diagnosis of sarcoidosis is played by histologic evidence on the basis of which additional tests can be used. We present a case of primary sarcoidosis of the nose and sinuses which involved the orbit and had non-specific symptoms in the sinonasal region. The diagnosis of sarcoidosis was made ultimately only after decalcification of sample material taken from the ethmoidal labyrinth. Interspersed among the bone trabeculae there were the typical epithelioid cellular granulomas composed of epithelioid cells with round nuclei and prominent nucleoli, Langhans'-type giant cells, and a tender rim of chronic inflammatory infiltrate in the periphery. After initial beneficial response to the administered cortisone therapy (40 mg prednisolone daily for 6 months and then a daily maintenance dose of 10 mg) the control examination showed that the peri- and retrobulbar infiltrate persisted and the eye symptoms recurred. We therefore increased the dose and proceeded to a pulse therapy (120 mg of urbason daily and a daily maintenance dose of 30 mg). At present the patient is still receiving this therapy which has reduced as a result the local manifestations but Cushing's syndrome has developed as a side effect.     

Donation after cardiac death in pediatric critical care.

OBJECTIVE: To describe the unmet need for pediatric organs, the history of donation after cardiac death (DCD), implementation of DCD policies in children's hospitals, and the current U.S. experience with DCD in children. DESIGN: Review of existing literature and national data regarding DCD. SETTING: Three children's hospitals and a national organ procurement network. PATIENTS: Nationwide review of pediatric candidates for transplantation and pediatric DCD donors. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Interest in DCD has greatly increased over the past several years due to limited organ availability for transplantation. Leading medical groups have evaluated and endorsed DCD, and more hospitals are offering DCD as part of end-of-life care options for dying patients and to increase donation. Children's hospitals need to evaluate this concept and develop ethically sound polices to meet the needs of patients and families. Preliminary reports regarding organ function from DCD donors are promising, and DCD is increasing. CONCLUSIONS: The widening gap between the need for organs and the availability of organs from brain-dead donors has led to a resurgence of both interest in and use of organs donated after cardiac death. Children's hospitals need to explore DCD as an option in select circumstances to serve grieving families who would like to donate and to increase organ availability for transplantation. DCD programs are dependent on input and support from critical care providers.     

Kocuria Species Peritonitis: Although Rare,We Have To Care

Kocuria species are found in the environment and on human skin. These micro-organisms are generally considered to be nonpathogenic saprophytes, rarely causing infection. However, the peritoneum has been reported to be a site of Kocuria infection. We reviewed all cases of peritonitis in peritoneal dialysis (PD) patients caused by Kocuria species that were reported in the worldwide literature. In total, 12 episodes of Kocuria species peritonitis have been reported in 9 PD patients. The median age of the patients was 62 years (range: 8 – 78 years). In the reported episodes, 4 different Kocuria species were isolated, with K. varians being the predominant species (41.7%). The most common initial symptom was abdominal pain (83.3%), followed by turbid effluent (75%) and fever (33.3%). Intraperitoneal first-generation cephalosporins and glycopeptides were the most-used antibiotics, with first-generation cephalosporins being more often preferred as first-line therapy. The median duration of treatment was 14 days, and in 2 episodes, the Tenckhoff catheter was removed. Although Kocuria peritonitis in PD patients is rare, it should be promptly treated because relapses can occur, especially with K. varians episodes.     

Development of a novel PTT assay for mutation detection in <Emphasis Type="Italic">PALB2</Emphasis> large exons and <Emphasis Type="Italic">PALB2</Emphasis> screening in medullary breast cancer

Beyond BRCA1 and BRCA2 genes, PALB2 (Partner and localizer of BRCA2) emerges as the third breast cancer susceptibility gene due to its role in the same DNA repair pathway: homologous recombination. In most populations studied so far, PALB2 mutations are detected in 1–2 % of BRCA negative female patients. PALB2 gene contains 13 exons; exons 4 and 5 consist 65 % of the coding area. We developed a protein truncation test (PTT) for quick screening of truncating pathogenic mutations of these two large exons. Specific primers were de novo, in silico designed and the PTT-PCR products were translated in the presence of biotinylated lysine and detected colorimetrically. The assay was initially tested in 30 patients with hereditary breast cancer, negative for BRCA mutations and then, in 17 patients with the rare medullary breast cancer subtype. Small PALB2 exons were screened with high-resolution melting curve analysis (HRMA) and the large DNA rearrangements with multiplex ligation-dependent probe amplification (MLPA). Any alterations detected were verified by Sanger DNA Sequencing. The developed PTT methodology is highly specific for clinical significant mutations; positive control samples that produce truncated PALB2 peptides were correctly identified and the method was accurate when compared to DNA sequencing. We did not detect any deleterious PALB2 mutation in both groups of patients. HRMA and MLPA were also negative for all tested samples. However, our novel, fast and cost-effective PTT method for pathogenic mutation detection of the two large PALB2 exons can be applied in screening of a large number of breast cancer patients.