Search

Background: Previously, we demonstrated that the ancestry‐related risk for Late Onset Alzheimer's Disease (LOAD) is driven by a local genomic region (termed Local Ancestry; LA) around APOEε4. Furthermore, we showed that in the brain of individuals bearing European LA there is higher expression of APOEε4 compared to those with African LA. In a follow‐up study, utilizing reporter assays and Capture‐C data we located two intronic regions within the European LA, both in the TOMM40 gene (named B10 and B13), that increased APOE expression in microglia and astrocytes. In this study, we sought to validate their regulatory role in APOE expression using CRISPR interference/activation (CRISPRi/a). Method: Human Microglial Clone 3 (HMC3) CRISPRi/a lines were produced by transducing inducible dCas9‐VP64 (Activation), dCas9‐KRAB (Interference) or dCas9 (control) using lentiviral vectors. To direct the dCas9 constructs to our regions of interest, we generated multiplex vectors that encode 4 short‐guide RNAs (sgRNAs) targeting either B10 or B13. We used 4 different sgRNAs in each case to ensure full‐length coverage of the tested regions (∼850bp size). An empty multiplex vector was used as a control. We then transduced either of the multiplex vectors into the HMC3 CRISPRi/a lines. We induced expression of the dCas9 constructs for 2 or 6 days with Doxycycline (2ug/ml). RNA was extracted and the expression of APOE and TOMM40 was measured by qRT‐PCR. Result: APOE expression significantly increased when targeting B10 or B13 (p=0.001; p=0.003 respectively) with dCas9‐VP64 after 2 days of Doxycycline treatment. Six days after treatment the significance persisted only when targeting B10 (p=0.01). No significant changes in APOE expression were observed in the cells bearing the dCas9‐KRAB presumably due to low endogenous APOE levels. Expression of TOMM40 did not vary under any treatment. Conclusion: These preliminary results support our previous findings that regions B10 and B13 may act as regulators for APOE expression as demonstrated by the elevation of ApoE expression when targeting an activator to these regions. The expression of TOMM40 did not vary across cell lines in the evaluated time points supporting that the effect observed is specific for APOE. [ABSTRACT FROM AUTHOR]

Background: The risk for late‐onset Alzheimer disease (AD) in APOEε4 carriers differs between ancestral groups, where APOEε4's odds ratio for AD risk is lower in African (AFR) homozygous carriers than in non‐Hispanic White (NHW) or Japanese (JPT) carriers (odds ratio ∼2‐5 vs >15). Local ancestry (LA) analyses in APOEε4 carrier populations have shown the protective effect in AFR relative to EUR/JPT is due to noncoding factors lying in the LA surrounding APOEε4. Thus, regulatory differences between risk and protective LA haplotypes are most likely involved in the differential risk effect seen for APOEε4 on different backgrounds. Methods: We identified 56 significant sequence differences between AFR and EUR/JPT APOEε4 haplotypes from the 1000 genomes in the immediate topologically associated domain surrounding APOE. We performed two different Massively Parallel Reporter Assay (MPRA) designs; one assessing small haplotype (∼900bp) effects and one based upon single variant effects. We supplemented these results with single fragment luciferase reporter assays. All assays were performed in at least duplicate in HMC3 (microglia), U118 (astrocytes) and SH‐SY5Y (neurons) cell lines. Additionally, we integrated chromatin interaction information from promoter capture C chromatin conformation assays in the same cell types. Results: We identified a region in the first introns of TOMM40 with increased EUR/JPT enhancer activity, supported by both MPRA analyses and APOE promoter interaction in astrocytes and microglia. Two additional regions with differential enhancer activity in neurons, but no promoter interaction, were identified; downstream of APOE and in PVRL2 introns upstream of APOE presenting with higher EUR or higher JPT haplotype variant enhancer activity compared to AFR, respectively. Conclusions: Our results indicate several areas of differential regulation in this LA region on APOEε4 haplotypes dependent on cell type. As APOE is mostly expressed in glial cells, the data in TOMM40 introns points to this region as having the biggest impact on APOE expression in our study and thus highly supports the involvement of this region in the differential risk effects seen for APOEε4. Follow‐up of the identified regulatory regions is currently ongoing using in‐house iPSC derived cell lines. [ABSTRACT FROM AUTHOR]

Background: Alzheimer's disease (AD) research has principally focused on neurons due to their role in neurodegeneration. In contrast, recent studies suggest that genetic mechanisms drive microglia to prolonged inflammation in AD brains, exacerbating neurodegeneration. Indeed, recent genome‐wide association studies (GWAS) of AD have identified multiple loci near genes related to microglial function, such as TREM2 and CR1. However, GWAS does not have the sensitivity to identify causal variants or effector genes. We used a combination of ATAC‐seq and high‐resolution promoter‐focused Capture‐C in two human microglial cell models to map interactions between GWAS‐implicated variants and their putative effector genes. We then validated an observed interaction at the 'CASS4' locus using CRISPR‐Cas9 genome editing. Method: Improving on the relatively low resolution of available Hi‐C data, we employed high resolution Capture‐C to characterize the physical genome‐wide interactions of all human promoters. We performed this in the human microglial cell line HMC3 and human iPSC‐derived microglia (iMg). We confirmed the enhancer activity of a SNP we elected to pursue and its associated regulatory element using dual‐luciferase assays. To confirm the regulatory interaction with the Capture‐C implicated effector gene, we used lentiviral CRISPR‐Cas9 to delete an approximately 300bp region containing the SNP in HMC3 cells. We then confirmed changes in RNA and protein expression using RNA‐seq and Western blotting, respectively. Result: Variant‐to‐gene mapping in both microglial cell models identified 67 putative effector genes (51 coding) across both cell types, with 14 observed in both models. We identified a novel proxy SNP, rs6024870 (r2 = 0.93 to sentinel SNP rs6014724), at the 'CASS4' locus, which coincided with open chromatin and directly contacted the promoter of RTFDC1, a gene not previously implicated in AD. Deletion of the putative enhancer region harboring rs6024870 by CRISPR‐Cas9 in HMC3 reduced the expression of RTFDC1 at both the mRNA and protein level. We also note that CASS4 levels were modestly influenced by this CRISPR‐mediated perturbation. Conclusion: We implicate RTFDC1 as a novel effector gene at the AD 'CASS4' GWAS locus. Further efforts will characterize the phenotypic effect of this variant in microglial cell models, including on inflammation and phagocytic activity. [ABSTRACT FROM AUTHOR]

Biomass utilization seems to be the easiest way to increase the exploitation of renewable energy sources for small and medium scale energy production. In particular, ligneous biomass, thanks to its availability, appears to be the most suitable for cogeneration applications. Among the several technologies available for energy conversion, the most frequently utilized system to generate electric power is still a process based on a Rankine or Hirn cycle with water as a working fluid. This solution seems quite interesting particularly for small size cogeneration plants because of its availability on the market and the analogy with typical applications involving fossil fuels. Many investors (e.g. Energy Service Companies, ESCOs) might be interested in the installation of this type of cogeneration plants in order to reduce the environmental impact of the system and therefore to benefit from governmental incentives. In order to assess the feasibility of this type of plants, it is important to investigate the possible thermodynamic, economic and technologic constraints. An analysis of these factors has been carried out in the present study. [ABSTRACT FROM AUTHOR]

The heart failure (HF) is one of the greatest problems of public health with increasing epidemiological importance. In the present study we analyzed a population of 299 patients, consecutively admitted to hospital, whose diagnosis of HF was verified retrospectively. In our analysis we considered underlying heart diseases, comorbidities, ejection fraction, presence of atrial fibrillation and pleural effusion, values of NT pro-BNP and causes of destabilization precipitating HF. The mean age of our population was 81 years. Patients with preserved systolic function were 145 (61% of the total, 59 male and 86 female). 166 patients (69% of the total) had hypertensive heart disease and 211 had hypertension (88% of the total). Patients with pleural effusion were 108 (46% of total). In the total population 102 patients (43%) had from 3 to 5 comorbidities, 169 patients (71%) had at least 2 comorbidities and only 4 patients (1.7%) had no comorbidities. The collected data highlight the complexity of patients with HF, often due to advanced age and a high number of comorbidities. [ABSTRACT FROM AUTHOR]

Due to the increasing age of the population the number of people suffering from chronic pain has significantly increased. People with chronic pain suffer from various diseases. Often this pain is not adequately controlled and is refractory, while its neuropathic component, which requires a different treatment, is perhaps underestimated compared to more properly nociceptive pain. The purpose of this study was to evaluate the presence of a neuropathic component in a cohort of 105 patients consecutively admitted to three Internal Medicine Units in Emilia Romagna. For the identification of the component of neuropathic pain diagnostic (DN4) questionnaire, previously validated, has been used. The average age of the patients studied was 64.4 years. The group of subjects with chronic non-cancer pain (78%) was numerically higher than the group of patients suffering from cancer pain (22%). All patients had pain and, according to the visual analogue scale (VAS), pain ranged from moderate to severe (median 7). Although without reaching statistically significant data, according to the VAS scale, cancer pain had an average higher value than non-cancer pain (7 vs 6.5). The prevalence of neuropathic component of pain was higher in patients with non-cancer pain (66% vs 57%). Instead, the recorded pain intensity in patients with neuropathic component was statistically much higher than the group in which the neuropathic component was absent (6.9 vs 6.1; P<0.05). In patients suffering from chronic pain, regardless of its nature and its etiology, the presence of a neuropathic component is significant. We must become aware of it and must search for it regularly through appropriate tools, such as the DN4 questionnaire. The presence of a neuropathic component usually makes the pain more intense and more refractory to treatments commonly used. Search for it may have therapeutic implications, suggesting that doctors use drugs active on this component. Since the majority of patients suffering from chronic pain are admitted to internal medicine wards, this awareness ought to become cultural heritage for the internist. [ABSTRACT FROM AUTHOR]

Copyright of Italian Journal of Medicine is the property of PAGEPress and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Italian Journal of Medicine is the property of PAGEPress and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Italian Journal of Medicine is the property of PAGEPress and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The article reports the results of an observational study conducted in 53 internal medicine departments of the Emilia Romagna region based on the answers to 54 questions administered in a questionnaire, submitted electronically, based on the type of center (large >250 beds, small <250 beds), laboratory equipment, management aspects, pharmacological and clinical/economic aspects and use of antibiotics in the empirical treatment of severe infections. The result is a snapshot of the existing situation which shows a substantial availability of resources and a good level of expertise in the field of infectious disease management, despite some areas still need improvement. It also highlights some differences in terms of procedures amongst large hospitals, where infectious diseases are treated by an ad hoc specialized staff, and the small ones, where the internist is generally involved, being responsible for both direct management of severe infections and giving advice to other departments (emergency or surgery). It emerged that, both in small and large hospitals, more discussion and continuous assessment are needed to apply the appropriate protocols based on reference checklists. [ABSTRACT FROM AUTHOR]

Genome-wide association studies (GWAS) have identified >300 loci associated with measures of adiposity including body mass index (BMI) and waist-to-hip ratio (adjusted for BMI, WHRadjBMI), but few have been identified through screening of the African ancestry genomes. We performed large scale meta-analyses and replications in up to 52,895 individuals for BMI and up to 23,095 individuals for WHRadjBMI from the African Ancestry Anthropometry Genetics Consortium (AAAGC) using 1000 Genomes phase 1 imputed GWAS to improve coverage of both common and low frequency variants in the low linkage disequilibrium African ancestry genomes. In the sex-combined analyses, we identified one novel locus (TCF7L2/HABP2) for WHRadjBMI and eight previously established loci at P < 5×10−8: seven for BMI, and one for WHRadjBMI in African ancestry individuals. An additional novel locus (SPRYD7/DLEU2) was identified for WHRadjBMI when combined with European GWAS. In the sex-stratified analyses, we identified three novel loci for BMI (INTS10/LPL and MLC1 in men, IRX4/IRX2 in women) and four for WHRadjBMI (SSX2IP, CASC8, PDE3B and ZDHHC1/HSD11B2 in women) in individuals of African ancestry or both African and European ancestry. For four of the novel variants, the minor allele frequency was low (<5%). In the trans-ethnic fine mapping of 47 BMI loci and 27 WHRadjBMI loci that were locus-wide significant (P < 0.05 adjusted for effective number of variants per locus) from the African ancestry sex-combined and sex-stratified analyses, 26 BMI loci and 17 WHRadjBMI loci contained ≤ 20 variants in the credible sets that jointly account for 99% posterior probability of driving the associations. The lead variants in 13 of these loci had a high probability of being causal. As compared to our previous HapMap imputed GWAS for BMI and WHRadjBMI including up to 71,412 and 27,350 African ancestry individuals, respectively, our results suggest that 1000 Genomes imputation showed modest improvement in identifying GWAS loci including low frequency variants. Trans-ethnic meta-analyses further improved fine mapping of putative causal variants in loci shared between the African and European ancestry populations.

Fever often occurs in cancer patients and the possibility of having a reliable marker for the differential etiological diagnosis is desirable. The aim of this study was to investigate the eligibility of the use of procalcitonin (PCT) in hemato-oncological patients for the differential diagnosis of fever. We prospectively enrolled 98 cancer patients and divided them into two groups: those with active disease and those with non-active disease. Procalcitonin was dosed at Time 0 (recruitment) and at the onset of fever. On enrollment, PCT values were 0.1 ng/mL in 83% patients with active disease, and lower than 0.5 ng/mL in 23%, which is usually considered not suggestive of bacterial infection. Four percent of patients had values over 0.5 ng/mL and these were mainly patients with neuroendocrine tumors or affiliates. On enrollment, there were also no statistically significant differences in PCT values between the two groups of patients. This showed that active cancer is unable by itself to change PCT levels. In the active disease tween group, 21 episodes of fever due to bacterial infection were registered, and in all of them an increase in PCT values was observed This demonstrates the ability of PCT to detect an infection-induced fever in cancer patients. Procalcitonin concentrations are not significantly altered by active neoplastic disease. On the contrary, in the course of fever due to a bacterial infection, PCT values increase and can, therefore, be considered a useful tool in the differential diagnosis between infection-induced fever and drug-related or tumor associated-fever. Procalcitonin may be a useful marker of bacterial infection even in cancer patients. [ABSTRACT FROM AUTHOR]

More and more frequently, patients admitted to surgical wards present characteristics similar to those admitted to medical units. They are fragile patients, often elderly, with significant comorbidity. In recent years, to address these emerging clinical issues in a surgical setting, different organizational models involving specialists of different backgrounds were studied, and in particular involving internists and geriatricians. To widen our current knowledge, in 2011 the Federation of Associations of Hospital Doctors on Internal Medicine of Emilia-Romagna, northern Italy (FADOI-ER), proposed a questionnaire to the public healthcare internal medicine departments of the Emilia Romagna region to collect information as to in what way and to what extent internists are involved in the management of surgical patients. In this article, we analyze the results of the questionnaire and make some organizational considerations and proposals. The questionnaire was very simple, consisting of 14 items. The survey was conducted from 1-28 February 2011. Replies were received from 20 internal medicine departments of a total of 75 in the Emilia Romagna region. The FADOI-ER survey has some limitations, the first of which is that only just under 25% of internal medicine departments in the Emilia Romagna region took part. However, the results are still interesting and seem to suggest that internists, because of their particular cultural background and training, could be the preferred partners for comanagement within the context of inpatient surgical procedures. The results of the FADOI-ER questionnaire are also consistent with the data reported in literature and daily clinical experience that highlight the need for a more multi-specialist approach to patient management with medical internists. Further studies will help provide answers as to the best way to conduct this multidisciplinary approach that could represent one of the future challenges for healthcare. [ABSTRACT FROM AUTHOR]

Abstract: To assist ongoing investigations of the limits of the tradeoff between spatial resolution and noise in PET imaging, several PET instruments based on silicon-pad detectors have been developed. The latest is a segment of a dual-ring device to demonstrate that excellent reconstructed image resolution can be achieved with a scanner that uses highresolution detectors placed close to the object of interest or surrounding a small field-of-view in combination with detectors having modest resolution at larger radius. The outer ring of our demonstrator comprises conventional BGO block detectors scavenged from a clinical PET scanner and located at a 500mm radius around a 50mm diameter field-of-view. The inner detector–in contrast to the high-Z scintillator typically used in PET–is based on silicon-pad detectors located at 70mm nominal radius. Each silicon detector has 512 1.4mm x 1.4mm x 1mm detector elements in a 16 x 32 array and is read out using VATA GP7 ASICs (Gamma Medica-Ideas, Northridge, CA). Even though virtually all interactions of 511 keV annihilation photons in silicon are Compton-scatter, both high spatial resolution and reasonable sensitivity appears possible. The system has demonstrated resolution of ∼ 0.7mm FWHM with Na-22 for coincidences having the highest intrinsic resolution (silicon-silicon) and 5–6mm FWHM for the lowest resolution BGO-BGO coincidences. Spatial resolution for images reconstructed from the mixed silicon-BGO coincidences is ∼1.5mm FWHM demonstrating the “magnifying-glass” concept. [Copyright &y& Elsevier]

Copyright of Italian Journal of Medicine is the property of PAGEPress and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract: Multiple-view L 2 triangulation is a key problem in computer vision. This paper addresses the standard case where all image points are available, and the case where some image points are not available. In the latter case, it is supposed that the unknown image point belongs to a known region such as a line segment or an ellipse, as it happens for instance due to occlusions. For this problem we propose two methods based on linear matrix inequalities (LMIs). The first method, named TFML, exploits the fundamental matrix and is fast (the average computational time with two and three-views is 0.01 and 0.05s on Matlab) at the expense of possible conservatism, which however it is shown to occur rarely in practice, and which can be immediately detected. The second method, named TPML, exploits the projection matrix, is slower, but allows one to reduce the conservatism by using techniques for optimization over polynomials. Various examples with synthetic and real data illustrate the proposed strategy. [ABSTRACT FROM AUTHOR]

The article discusses a study which detailed an osteological description of the shell of Testudo kleinmanni. Testudo kleinmanni is a small tortoise species found only in a narrow, discontinuous strip along the southeastern Mediterranean coast that extends from Libya to Israel. The characteristics of its nuchal, suprapygal, pygal, epiplastron, entoplastron, hyoplastron and xiphiplastron are said to differentiate it from Testudo graeca. According to the study authors, the rarity of fossil and subfossil records relating to Testudo kleinmannu can be blamed to the fact that diagnostic osteological characters were previously unknown.

Background: Previous studies of Alzheimer's Disease (AD) have primarily implicated neurons and microglia as disease‐conferring cell types due to obvious links with degeneration and inflammation, respectively. However, astrocytes may also play a role in AD pathogenesis, but have been less studied in this context. Indeed, genome‐wide association studies (GWAS) have identified multiple genomic variants that reside near genes with astrocyte‐related functions, such as lipid processing and synaptic function. However, GWAS only reports genomic variants associated with a given trait and not necessarily the precise localization of culprit genes. High resolution chromatin conformation capture‐based techniques detect 3D genomic interactions between GWAS‐implicated signals and their effector genes, and allow for the characterization of non‐coding variants in the context of their regulatory activity. Method: To improve on the low resolution of available Hi‐C data, we employed a high resolution DpnII‐based Capture‐C method to simultaneously characterize the physical genome‐wide interactions of 36,691 baited regions covering human promoters genome‐wide (including non‐coding transcripts) ‐ which we applied to a human primary astrocyte line (NHA, Lonza). Following sequencing, we investigated significant interactions at two different resolutions (1‐fragment and 4‐fragment resolutions; median fragment size = 265bp and 1,441bp, respectively). In parallel, we generated ATAC‐seq open chromatin maps to filter for informative (r2>0.7) proxy single nucleotide polymorphisms (SNPs) for each of the 45 AD independent signals reported to date. Result: ATAC‐seq fine‐mapping yielded 100 candidate SNPs in open chromatin for 30 of these loci. By further constraining on our promoter Capture C data in astrocytes, at both one and four DpnII fragment resolution (median distance between interacting regions 9kb and 71kb, respectively), we observed contacts to "open" promoters for 6 putative target genes, corresponding to 6 of the original GWAS loci. These included PICALM, FERMT2, CASS4, and CLU. Conclusion: We observed informative contacts between proxy SNPs and putative effector genes in the human astrocyte context for ∼13% of AD GWAS loci. Further efforts in other relevant cell types, where many of the other loci may in fact be principally operating, should shed light on additional signals. Follow‐up functional studies are warranted to validate these findings. [ABSTRACT FROM AUTHOR]