Low-cost dedicated mini-arrays for high-throughput analysis of DNA copy-number alterations in neuroblastoma

ArrayCGH is commonly used for high-resolution detection of copy-number alterations in tumours, allowing identification of chromosomal aberrations with prognostic or diagnostic relevance. Currently available arrayCGH platforms are still very expensive for analysis of large sets of samples. For this purpose, we have constructed a dedicated mini-array that is enriched for BAC/PAC clones in the prognostic important regions for neuroblastoma and that only covers a small area on the slide, allowing down-scaling of the labelling and hybridisation reagents and hence reducing the price. The mini-arrays were validated on neuroblastoma samples and comparison with high-resolution whole-genome arrayCGH data yielded complete concordant results.     

Clinical Aspects of Palliative Sedation in Prospective Studies. A Systematic Review

ContextNear the end of life when patients experience refractory symptoms, palliative sedation may be considered as a last treatment. Clinical guidelines have been developed, but they are mainly based on expert opinion or retrospective chart reviews. Therefore, evidence for the clinical aspects of palliative sedation is needed.ObjectivesTo explore clinical aspects of palliative sedation in recent prospective studies.MethodsSystematic review was conducted following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines and registered at PROSPERO. PubMed, CINAHL, Cochrane, MEDLINE, and EMBASE were searched (January 2014–December 2019), combining sedation, palliative care, and prospective. Article quality was assessed.ResultsTen prospective articles were included, involving predominantly patients with cancer. Most frequently reported refractory symptoms were delirium (41%–83%), pain (25%–65%), and dyspnea (16%–59%). In some articles, psychological and existential distress were mentioned (16%–59%). Only a few articles specified the tools used to assess symptoms. Level of sedation assessment tools were the Richmond Agitation Sedation Scale, Ramsay Sedation Scale, Glasgow Coma Scale, and Bispectral Index monitoring. The palliative sedation practice shows an underlying need for proportionality in relation to symptom intensity. Midazolam was the main sedative used. Other reported medications were phenobarbital, promethazine, and anesthetic medication—propofol. The only study that reported level of patient's discomfort as a palliative sedation outcome showed a decrease in patient discomfort.ConclusionAssessment of refractory symptoms should include physical evaluation with standardized tools applied and interviews for psychological and existential evaluation by expert clinicians working in teams. Future research needs to evaluate the effectiveness of palliative sedation for refractory symptom relief.     

Familial pericentric inversion of chromosome 18: behavioral abnormalities in patients heterozygous for either the dup(18p)/del(18q) or dup(18q)/del(18p) recombinant chromosome

We describe a family in which the largest hitherto reported pericentric inversion of chromosome 18, inv(18)(p11.22q23), segregates. Individuals heterozygous for the nonrecombinant inversion were unaffected. However, those heterozygous for either the dup(18p)/del(18q) or dup(18q) /del(18p) recombinant exhibited mild learning difficulty, personality disorders and deficient social behavior in the absence of mental retardation. Of the three family members tested, the behavioral abnormalities were more prominent in the two individuals with the dup(18p)/del(18q) recombinant than in the one with the dup(18q)/del(18p) recombinant. Genetic counseling issues for this family, in particular for the affected, include the enhanced probability of reduced fertility as well as the recurrence risk of the parental inversion equaling 1/2 in surviving offspring. This observation kindles the interest in determining the frequency of subtelomeric rearrangements in individuals with learning difficulty and deficiency in social interaction, phenotypic features often considered to be of multifactorial causation.     

Differential induction of monocyte chemotactic protein-3 in mononuclear leukocytes and fibroblasts by interferon-alpha/beta and interferon-gamma reveals MCP-3 heterogeneity

Monocyte chemotactic protein-3 (MCP-3) is a pluripotent CC chemokine, attracting most leukocytic cell types. With the use of a sensitive and specific ELISA, MCP-3 was found to be inducible in fibroblasts and peripheral blood mononuclear cells (PBMC) by cytokines and cytokine inducers. MCP-3 production levels (1-10 ng/ml) were tenfold lower compared to those of MCP-1. In diploid fibroblasts, synergistic induction of MCP-3, but not of MCP-1, mRNA and protein was observed by combined treatment with IL-1beta and IFN-gamma. In PBMC, IFN-alpha and IFN-beta (but not IFN-gamma), as well as measles virus and double-stranded RNA, were potent inducers of MCP-3, which suggests a role for this chemokine in an early stage of viral infections. In contrast, endotoxin failed to induce MCP-3 production in fibroblasts and PBMC. Purification of MCP-3 from PBMC revealed biochemical heterogeneity. In monocyte chemotaxis and calcium mobilization assays, pure 11-kDa MCP-3 from PBMC showed similar potencies as MCP-3 from tumor cells. It was concluded that the induction of MCP-3 by IFN is regulated differently in fibroblasts and PBMC. In view of the multiple target cells for MCP-3, local and strictly regulated chemokine production might be important to conduct selectively the immune response in infection or inflammation.     

Fourteen new cases contribute to the characterization of the 7q11.23 microduplication syndrome

Interstitial deletions of 7q11.23 cause Williams–Beuren syndrome, one of the best characterized microdeletion syndromes. The clinical phenotype associated with the reciprocal duplication however is not well defined, though speech delay is often mentioned. We present 14 new 7q11.23 patients with the reciprocal duplication of the Williams–Beuren syndrome critical region, nine familial and five de novo. These were identified by either array-based MLPA or by array-CGH/oligonucleotide analysis in a series of patients with idiopathic mental retardation with an estimated population frequency of 1:13,000–1:20,000. Variable speech delay is a constant finding in our patient group, confirming previous reports. Cognitive abilities range from normal to moderate mental retardation. The association with autism is present in five patients and in one father who also carries the duplication. There is an increased incidence of hypotonia and congenital anomalies: heart defects (PDA), diaphragmatic hernia, cryptorchidism and non-specific brain abnormalities on MRI.Specific dysmorphic features were noted in our patients, including a short philtrum, thin lips and straight eyebrows. Our patient collection demonstrates that the 7q11.23 microduplication not only causes language delay, but is also associated with congenital anomalies and a recognizable face.     

Osteopoikilosis, short stature and mental retardation as key features of a new microdeletion syndrome on 12q14

This report presents the detection of a heterozygous deletion at chromosome 12q14 in three unrelated patients with a similar phenotype consisting of mild mental retardation, failure to thrive in infancy, proportionate short stature and osteopoikilosis as the most characteristic features. In each case, this interstitial deletion was found using molecular karyotyping. The deletion occurred as a de novo event and varied between 3.44 and 6 megabases (Mb) in size with a 3.44 Mb common deleted region. The deleted interval was not flanked by low-copy repeats or segmental duplications. It contains 13 RefSeq genes, including LEMD3, which was previously shown to be the causal gene for osteopoikilosis. The observation of osteopoikilosis lesions should facilitate recognition of this new microdeletion syndrome among children with failure to thrive, short stature and learning disabilities.     

4q34.1–q35.2 deletion in a boy with phenotype resembling 22q11.2 deletion syndrome

Small terminal or interstitial deletions involving bands 4q34 and 4q35 have been described in several patients with a relatively mild phenotype such as mild to moderate intellectual disability and minor dysmorphic features. We present a boy born from unrelated parents with a de novo 4q34.1–q35.2 deletion and clinical features resembling 22q11.2 deletion syndrome. To the best of our knowledge, this is the first reported patient with 4q34–q35 deletion and phenotype resembling 22q11.2 deletion syndrome without fifth finger anomalies as a specific feature of 4q- syndrome. G-banding karyotyping disclosed the deletion, which was further delineated by microarray comparative genomic hybridization. Fluorescence in situ hybridization and multiplex ligation-dependent probe amplification analyses did not reveal rearrangements of 22q11.2 region. MLPA confirmed the deletion within the 4q35.2 region. Conclusion: Given the considerable clinical overlaps between the 22q11.2 deletion syndrome and clinical manifestation of the patient described in this study, we propose that region 4q34.1–q35.2 should be considered as another region associated with phenotype resembling 22q11.2 deletion syndrome. We also propose that distal 4q deletions should be considered in the evaluation of patients with phenotypic manifestations resembling 22q11.2 deletion syndrome in whom no 22q11.2 microdeletion was detected, even in the absence of distinctive fifth finger anomalies. Additionally, we underline the importance of applying array CGH that enables simultaneous genome-wide detection and delineation of copy number changes (e.g., deletions and duplications).     

Emergency hospital admission for pain in palliative patients: A crucial role for general practitioners

Abstract

Introduction: In previous papers we analysed the incidence of wheezing in young children living near the iron and steel factory in C?l?ra?i, and in a control region (Rose?i) without industrial air pollution. Research question: Is industrial air pollution exposure during the first years of life a risk factor for the presence of asthmatic symptoms in children at school age? Methods: We assessed the prevalence of asthma symptoms using the ISAAC short form questionnaire for children aged 6–7 years, and measured the FEV1 and PEF in the children of both municipalities (297 children in C?l?ra?i, i.e. the exposed cohort, and 237 in Rose?i, i.e. the non-exposed cohort). Results: We found an OR of 7.2 (95% CI: 3.6–14.3) for affirmative answers to at least one of ISAAC questions for children living in C?l?ra?i compared to the children in Rose?i. The numbers of affirmative answers to all but one of the ISAAC questions were significantly higher in C?l?ra?i. The main result remained robust after adjusting for a series of co-variables using multiple logistic regression analysis (OR 14.8; 95% CI: 4.8–46.1). There was a strong relation between early life wheezing and asthma symptoms at school age (OR 9.0; 95%CI: 3.4–23.5).

Conclusion: Children, who had been living near an iron and steel factory during their early years, are still at increased risk for asthma symptoms at school age.     

Non-invasive grading of brain tumours using dynamic amino acid PET imaging : does it work for <Superscript>11</Superscript>C-Methionine?

BACKGROUND: Static imaging of amino acids does not allow differentiation of low versus high grade brain tumours. It has been shown that dynamic imaging of the amino acid analogue (18)F-fluoroethyltyrosine (FET) can achieve this goal. In many centres, (11)C-methionine (MET) is used for tumour imaging, but no clinical studies on the use of dynamic scanning for grading have been performed. METHODS: Thirty-four patients with primary brain glioma and histopathological confirmation were retrospectively studied using 40 min dynamic MET-PET with 220 MBq 11C-methionine. In relation to histopathological grading, various metabolic indices and temporal parameters as documented by Poepperl et al. (JNM 2006;47:393-403) were analyzed. RESULTS: None of the evaluated static or temporal parameters allowed discrimination between high and low grade tumours. On average, low grade tumours showed washout after the initial uptake maximum, while both increases and decreases were seen for high grade tumours. Only the relative early versus late uptake ratio showed a trend towards significance (-0.16 +/- 0.17 for low grade versus 0.01 +/- 0.25 for high grade; p = 0.07). CONCLUSION: Unlike FET-PET, the uptake characteristics of MET-PET do not allow classification of low and high grade tumours on an individual patient basis. Since literature data indicate that both tracers have a similar performance regarding biopsy location, tumour delineation, and detection of recurrence, FET-PET should be advocated over MET-PET as its uptake mechanism also allows noninvasive grading in glioma.