Hooked Up from a Distance: Charting Genome-Wide Long-Range Interaction Maps in Neural Cells Chromatin to Identify Novel Candidate Genes for Neurodevelopmental Disorders.

DNA sequence variants (single nucleotide polymorphisms or variants, SNPs/SNVs; copy number variants, CNVs) associated to neurodevelopmental disorders (NDD) and traits often map on putative transcriptional regulatory elements, including, in particular, enhancers. However, the genes controlled by these enhancers remain poorly defined. Traditionally, the activity of a given enhancer, and the effect of its possible alteration associated to the sequence variants, has been thought to influence the nearest gene promoter. However, the obtainment of genome-wide long-range interaction maps in neural cells chromatin challenged this view, showing that a given enhancer is very frequently not connected to the nearest promoter, but to a more distant one, skipping genes in between. In this Perspective, we review some recent papers, who generated long-range interaction maps (by HiC, RNApolII ChIA-PET, Capture-HiC, or PLACseq), and overlapped the identified long-range interacting DNA segments with DNA sequence variants associated to NDD (such as schizophrenia, bipolar disorder and autism) and traits (intelligence). This strategy allowed to attribute the function of enhancers, hosting the NDD-related sequence variants, to a connected gene promoter lying far away on the linear chromosome map. Some of these enhancer-connected genes had indeed been already identified as contributive to the diseases, by the identification of mutations within the gene's protein-coding regions (exons), validating the approach. Significantly, however, the connected genes also include many genes that were not previously found mutated in their exons, pointing to novel candidate contributors to NDD and traits. Thus, long-range interaction maps, in combination with DNA variants detected in association with NDD, can be used as "pointers" to identify novel candidate disease-relevant genes. Functional manipulation of the long-range interaction network involving enhancers and promoters by CRISPR-Cas9-based approaches is beginning to probe for the functional significance of the identified interactions, and the enhancers and the genes involved, improving our understanding of neural development and its pathology.

Intra-arterial chemotherapy for retinoblastoma: the dosimetric impact.

PURPOSE: Purposes are (1) to measure main radiation parameters and (2) to propose a method to estimate the absorbed doses of internal organs starting from DAP values. Measuring the exposition of internal organs by repeated irradiations on an anthropomorphic phantom with the same settings used in vivo, we could establish correlations between (1) DAP and the dose recorded by a dosimeter placed along the X-ray beam entrance pathway; (2) the dose recorded by the same dosimeter and the absorbed dose in internal organs. METHODS: Forty-four consecutive patients (16 males, 28 females) (mean age 35.4 months) treated at our institution with IAC (216 procedures: 196 via the ICA and 20 into branches of the ECA) were included in this prospective study. IAC was divided into 5 phases. Fluoroscopic time, DAP, and ESD were measured. RESULTS: The mean DAP was 595 ± 445 cGy cm2 and the mean fluoroscopic time was 540 ± 403 s. ESD was on average 9.59 mGy (range 0.8-165 mGy). The absorbed dose was lower than 12.1 mGy in the left retina (the more exposed organ) in 75% of single treatments and lower than 25 mGy in 95% of treatments. In the cases of 3 and 6 sessions, the left retina of 75% of patients absorbed respectively less than 36.3 and 72.7 mGy, whereas the left retina of 95% of patients received less than 75.2 and 150.4 mGy. Other organs were less exposed. CONCLUSION: This paper describes a method of absorbed dose estimation providing ranges used clinically in a single practice and the basis for further prospective studies.

Voltage-Gated Potassium Channel Antibodies in Slow-Progression Motor Neuron Disease.

BACKGROUND: The spectrum of autoimmune neurological diseases associated with voltage-gated potassium channel (VGKC)-complex antibodies (Abs) ranges from peripheral nerve disorders to limbic encephalitis. Recently, low titers of VGKC-complex Abs have also been reported in neurodegenerative disorders, but their clinical relevance is unknown. OBJECTIVE: The aim of the study was to explore the prevalence of VGKC-complex Abs in slow-progression motor neuron disease (MND). METHODS: We compared 11 patients affected by slow-progression MND with 9 patients presenting typical progression illness. Sera were tested for VGKC-complex Abs by radioimmunoassay. The distribution of VGKC-complex Abs was analyzed with the Mann-Whitney U test. RESULTS: The statistical analysis showed a significant difference between the mean values in the study and control groups. A case with long-survival MND harboring VGKC-complex Abs and treated with intravenous immunoglobulins is described. CONCLUSION: Although VGKC-complex Abs are not likely to be pathogenic, these results could reflect the coexistence of an immunological activation in patients with slow disease progression.

Adverse haematological outcome and environmental lead poisoning.

The South-eastern Area (SA) of the Municipality of La Spezia (Liguria Region, Italy) is characterised by a heavy environmental lead (Pb) contamination, chiefly due to the emissions of a Pb-processing plant in operation since 1930. In order to assess the risk of Pb poisoning of residents of SA, and to estimate the degree of association between the blood Pb level (BLL) and haematocrit % (HCT), intended as a biomarker of early haematological dysfunction, we reanalysed data of 785 individuals collected in 1992 as a part of a larger national biological monitoring project. Multiple normal regression modelling was applied to estimate the role of residence on log-transformed BLL, and Median Ratio (MR) was used as an index of effect. The same statistical modelling was also applied to reveal the relationship between HCT and BLL. Allowing for several confounders (including occupational exposure to Pb), residents of SA showed a 14% increase (MR=1.14, 95% IC=1.06-1.23%) in the median BLL value compared to people living outside SA. The excess reached 27% (MR=1.27, 95% IC=1.14-1.41%) after 30 years of residence. Parallel results were also obtained in a subgroup composed only of pupils (<18 years, non-smokers, non-drinkers). Finally, regression analysis highlighted a statistically significant parabolic trend in HCT in relation to BLL. The non-linear dose-response relationship, which attests to an adverse effect on the erythrocytic function of BLLs at least over 17.00 microg/dl, is in agreement with the findings of other authors and consistent with the results of an excess occurrence of self-reported anaemia obtained from a previous comparative survey carried out on the same population.

Lung cancer mortality in a district of La Spezia (Italy) exposed to air pollution from industrial plants.

AIMS AND BACKGROUND: In the last decades, many epidemiological studies have implicated outdoor environmental carcinogens in the onset of lung cancer. The present investigation evaluated lung cancer mortality in two areas of the Province of La Spezia (Northern Italy) exposed to environmental pollution emitted by a coal-fired power station and other industrial sources, including a waste incinerator. METHODS: In the two exposed areas, lung cancer mortality risk for the 1988-1996 calendar period was evaluated using the whole Province population as referent. The corresponding relative risks (RR) were estimated after controlling for age structure, urban/rural gradient and deprivation factors (occupation, education, home ownership, housing conditions and family structure) by a Poisson regression modeling. The geographic pattern of risk for the whole province was evaluated via the Besag, York and Mollié (BYM) bayesian model. RESULTS: Persons living in urban areas showed the highest rates in both sexes. No statistically significant risk excess was found in the two exposed areas among males, after excluding rural and semi-rural zones from the analyses (RR = 1.03 and RR = 0.77). In contrast, a risk excess was observed for females in both exposed areas, which remained elevated and statistically significant (P <0.05) after restriction to urban/semi-urban municipalities and after controlling for deprivation factors (RR = 1.54 and RR = 2.14, respectively). Bayesian mapping confirmed the rural/urban gradient and the risk excess observed in females near the industrial sites. CONCLUSIONS: The risk observed among females is consistent with pollution measurements and with other epidemiologic findings, whereas a strong confounding from occupational exposures and smoking habit could account for the lack of an excess risk in males. However, the ecologic nature of this investigation prevented drawing a causal inference. The pollution-related risk observed in the female gender is an important clue that deserves further epidemiologic attention.

[Lead pollution and hematologic effects in the city of La Spezia]. / Inquinamento da piombo ed effetti ematologici nel Comune di La Spezia.

During 1998-1999 a survey assessed the health impact of living in an area (Zona D) of the city of La Spezia (Liguria Region, Italy) characterised by industrial plants, urban waste disposal sites and incinerators, a coal-fired power-station and a heavily trafficked thoroughfare. One of the main results was an excess prevalence and incidence of self-reported anaemia among residents in area D compared to those of the control area (Zone C). Given the environmental lead pollution in Zone D due to the emissions of a lead processing plant active since 1930, and in order to confirm the validity of previous findings, was investieable the association between residence and blood lead levels, as well as the relationship between the latter and an anaemia-specific biomarker. We re-analysed personal, occupational and haematological data of 785 residents of La Spezia Province collected by the Liguria Environmental Protection Agency (ARPAL) during 1992 as part of a biological monitoring survey of the general population about the risk of lead intoxication. Subjects were geo-referenced by the 1992 residence address, and the haematocrit value was used as the anaemia-specific biomarker. Multiple regression analysis estimated the effect of residence on log-transformed lead blood levels, and to assess the relationship between the latter and haematocrit values. After adjusting for several potential confounding factors, residents from Zone D showed a 14% increase (95% IC = 6%-23%) in the blood lead median value estimated via the log-normal regression model compared to people living elsewhere. The excess became 27% (95% IC = 14%-41%) after 30 years of residence in the study area. Similar results were obtained by focusing analyses on students (n = 89, < 18 years, non-smokers, non-drinkers). The same modelling highlighted a significant non-linear (parabolic) dose-response relationship between lead blood and haematocrit values. This trend is consistent with the findings of other Authors and might reveal an adverse haematological effect of lead values over 170, 0 microgram/l. In conclusion, The results of this investigation strengthen the epidemiological evidence of the 1998-1999 survey.