DNA methylation profiling reveals common signatures of tumorigenesis and defines epigenetic prognostic subtypes of canine Diffuse Large B-cell Lymphoma

Epigenetic deregulation is a hallmark of cancer characterized by frequent acquisition of new DNA methylation in CpG islands. To gain insight into the methylation changes of canine DLBCL, we investigated the DNA methylome in primary DLBCLs in comparison with control lymph nodes by genome-wide CpG microarray. We identified 1,194 target loci showing different methylation levels in tumors compared with controls. The hypermethylated CpG loci included promoter, 5'-UTRs, upstream and exonic regions. Interestingly, targets of polycomb repressive complex in stem cells were mostly affected suggesting that DLBCL shares a stem cell-like epigenetic pattern. Functional analysis highlighted biological processes strongly related to embryonic development, tissue morphogenesis and cellular differentiation, including HOX, BMP and WNT. In addition, the analysis of epigenetic patterns and genome-wide methylation variability identified cDLBCL subgroups. Some of these epigenetic subtypes showed a concordance with the clinical outcome supporting the hypothesis that the accumulation of aberrant epigenetic changes results in a more aggressive behavior of the tumor. Collectively, our results suggest an important role of DNA methylation in DLBCL where aberrancies in transcription factors were frequently observed, suggesting an involvement during tumorigenesis. These findings warrant further investigation to improve cDLBCL prognostic classification and provide new insights on tumor aggressiveness

Relationship between low Ankle-Brachial Index and rapid renal function decline in patients with atrial fibrillation: A prospective multicentre cohort study

OBJECTIVE: To investigate the relationship between Ankle-Brachial Index (ABI) and renal function progression in patients with atrial fibrillation (AF). DESIGN: Observational prospective multicentre cohort study. SETTING:Atherothrombosis Center of I Clinica Medica of 'Sapienza' University of Rome; Department of Medical and Surgical Sciences of University Magna Græcia of Catanzaro; Atrial Fibrillation Registry for Ankle-Brachial Index Prevalence Assessment-Collaborative Italian Study. PARTICIPANTS: 897 AF patients on treatment with vitamin K antagonists. MAIN OUTCOME MEASURES: The relationship between basal ABI and renal function progression, assessed by the estimated Glomerular Filtration Rate (eGFR) calculated with the CKD-EPI formula at baseline and after 2 years of follow-up. The rapid decline in eGFR, defined as a decline in eGFR >5 mL/min/1.73 m(2)/year, and incident eGFR<60 mL/min/1.73 m(2) were primary and secondary end points, respectively. RESULTS: Mean age was 71.8±9.0 years and 41.8% were women. Low ABI (ie, ≤0.90) was present in 194 (21.6%) patients. Baseline median eGFR was 72.7 mL/min/1.73 m(2), and 28.7% patients had an eGFR60 mL/min/1.73 m(2), 153 (23.9%) had a reduction of the eGFR <60 mL/min/1.73 m(2). ABI ≤0.90 was also an independent predictor for incident eGFR<60 mL/min/1.73 m(2) (HR 1.851, 95% CI 1.205 to 2.845, p=0.005). CONCLUSIONS: In patients with AF, an ABI ≤0.90 is independently associated with a rapid decline in renal function and incident eGFR<60 mL/min/1.73 m(2). ABI measurement may help identify patients with AF at risk of renal function deterioration

Frequency of left ventricular hypertrophy in non-valvular atrial fibrillation

Left ventricular hypertrophy (LVH) is significantly related to adverse clinical outcomes in patients at high risk of cardiovascular events. In patients with atrial fibrillation (AF), data on LVH, that is, prevalence and determinants, are inconsistent mainly because of different definitions and heterogeneity of study populations. We determined echocardiographic-based LVH prevalence and clinical factors independently associated with its development in a prospective cohort of patients with non-valvular (NV) AF. From the "Atrial Fibrillation Registry for Ankle-brachial Index Prevalence Assessment: Collaborative Italian Study" (ARAPACIS) population, 1,184 patients with NVAF (mean age 72 \ub1 11 years; 56% men) with complete data to define LVH were selected. ARAPACIS is a multicenter, observational, prospective, longitudinal on-going study designed to estimate prevalence of peripheral artery disease in patients with NVAF. We found a high prevalence of LVH (52%) in patients with NVAF. Compared to those without LVH, patients with AF with LVH were older and had a higher prevalence of hypertension, diabetes, and previous myocardial infarction (MI). A higher prevalence of ankle-brachial index 640.90 was seen in patients with LVH (22 vs 17%, p = 0.0392). Patients with LVH were at significantly higher thromboembolic risk, with CHA2DS2-VASc 652 seen in 93% of LVH and in 73% of patients without LVH (p &lt;0.05). Women with LVH had a higher prevalence of concentric hypertrophy than men (46% vs 29%, p = 0.0003). Logistic regression analysis demonstrated that female gender (odds ratio [OR] 2.80, p &lt;0.0001), age (OR 1.03 per year, p &lt;0.001), hypertension (OR 2.30, p &lt;0.001), diabetes (OR 1.62, p = 0.004), and previous MI (OR 1.96, p = 0.001) were independently associated with LVH. In conclusion, patients with NVAF have a high prevalence of LVH, which is related to female gender, older age, hypertension, and previous MI. These patients are at high thromboembolic risk and deserve a holistic approach to cardiovascular prevention

Full penetrance of Morgagni-Stewart-Morel syndrome in a 75-year-old woman: case report and review of the literature

Morgagni-Stewart-Morel syndrome is defined as the presence of hyperostosis frontalis interna, variably associated with metabolic, endocrine, and neuropsychiatric disorders. The possible cause-effect relationship of these associations remains uncertain. A 75-year-old woman presented with severe frontal headache and a history of psychotic disorders. On instrumental examination she was found to have extensive frontal hyperostosis and cortical atrophy. These findings, associated to the metabolic and neuropsychiatric pattern of the patient, are consistent with a high penetrance of Morgagni-Stewart-Morel syndrome. In this clinical case seminar, we summarize the current understanding of the association between hyperostosis frontalis interna and Morgagni-Stewart-Morel, based on a MEDLINE search (case reports, original articles, and reviews published between 1928 and 2011) on this topic. Possible pathophysiological mechanisms underlying both the headache and the hyperostosis frontalis interna are discussed. A case of full penetrance of Morgagni-Stewart-Morel syndrome is reported, presenting many of the clinical features described in the literature. Metabolic and endocrine dysfunctions should be interpreted not only as isolated components of the syndrome, but also as the reason behind its pathogenesis. Endocrine or nutritional disorders may have led to an altered bone metabolism with frontal bone apposition. On the other hand, the severity of our patient's neurological and psychiatric symptoms correlates well with the severity of her hyperostosis frontalis interna and the cortical atroph

HYPOTHYROIDISM IN THE ELDERLY:DIAGNOSTIC PITFALLS ILLUSTRATED BY A CASE REPORT

A diagnosis of hypothyroidism in the elderly can easily be overlooked if we rely exclusively on its clinical presentation because this may be highly non-specific, since the signs and symptoms of the disease are common to other diseases typical of old age, and even to the normal aging process. Imaging diagnostics (ultrasound or CT), when considered alone, are also of little use for the purpose of clarifying thyroid gland function. We report here on a case of primary hypothyroidism that was diagnosed late because the correlated signs and symptoms (asthenia, bradycardia, pleural effusions, hyponatremia, worsening renal and respiratory insufficiency, hoarseness) had previously been attributed to the normal aging process and to the patient's other health conditions (Parkinson's disease, PD; chronic obstructive pulmonary disease, COPD). After a couple of weeks of treatment with levothyroxine and liothyronine, there were clinical and laboratory evidences of an improvement in the patient's condition. She became more reactive, with a shriller voice. The pleural effusion disappeared, and so did the bradycardia. Laboratory tests showed normal sodium levels, and the renal insufficiency had improved. The lack of specificity of the clinical presentation of hypothyroidism in the elderly might justify the routine measurement of thyroid-stimulating hormone in these patients

A case of naturally evolving gout in an elderly man

Gout is a common disorder in adults that can lead to severe organ decline, disability and impaired quality of life due to the formation of periarticular tophi. We report a case of massive tophaceous gout in a 78-year-old man with a 16-year-long history of untreated disease. The patient gradually became disabled, his renal function deteriorated, and he finally died of sepsis. Our case demonstrates that chronic gout not only affects the joints, but is also associated with organ function decline and can, even nowadays, lead to death

Regulation of bovine CYP3A28 promoter by pregnane X receptor (PXR) and constitutive androstane receptor (CAR).

INTRODUCTION The regulation of cytochrome P450 3A (CYP3A) has been extensively studied in human liver. In cattle, some information on the expression and modulation of CYP3As are now available, but molecular mechanisms involved in basal and xenobiotic- mediated gene regulation are still unknown. In this study, selected transcription factor binding sites were studied for their role in the basal and nuclear receptor-driven trans-activation of CYP3A28 promoter. MATERIALS AND METHODS Putative binding sites for transcription factors were predicted within the CYP3A28 promoter region (~10 kbp) through specific software. Fragments of the CYP3A28 promoter were amplified from bovine liver gDNA and cloned into pGL4.10 vector. Bovine nuclear receptor (bPXR, bCAR and bRXRa) fulllength cDNAs were amplified and inserted into pCI-neo expression vector. Deletion of ER6, HNF-1 and HNF-4 cis-elements in the proximal promoter (PP) construct, and mutagenesis of both ER6 and HNF-4 binding elements within the PP region, and of DR5 within the Fragment 3 (F3) were performed through inverse PCR and according to Fang et al. (2012), respectively. To identify the most important regions for gene activation, cotransfection studies in HepG2 cells were performed using all the promoter constructs, bPXR and its agonist SR12813, or constitutively active bCAR. To assay the role of specific binding sites on CYP3A28 regulation, C3A cells were then co-transfected with deleted or mutated plasmids, bPXR and SR12813 or rifampicin (RIF), and bCAR. Human 3A4-XREM- luc and PBREM-tk-luc were used as positive controls. RESULTS AND CONCLUSIONS CYP3A28 promoter screening revealed PP and PP-F3 as the most important elements for bPXR and bCAR activation. 3A4-XREMluc, the PP and PP-F3 constructs were significantly activated by bPXR and SR12813 (P < 0.05), but not by RIF. Deletion of ER6, HNF-1 and HNF-4 lead to a significant decrease of PP activation by bPXR and SR12813 (P < 0.01), while the mutated single elements ER6, HNF-4 and DR5 did not. Conversely, only PP-F3 was significantly activated by bCAR (P < 0.05). Thus, CYP3A28 PP contains interaction sites for bPXR trans-activation. ER6 and HNF-1 elements seem to contribute to SR12813 response. F3 fragment is involved in bPXR- and bCAR-mediated CYP3A28 regulation, but additional binding elements, apart from DR5, need to be investigated. ACKNOWLEDGEMENTS Project supported by University of Padova (CPDA109434, 60A08-4783/14)