EGFR mutation analysis on circulating free DNA in NSCLC: a single-center experience.

PURPOSE: Monitoring mutation status in circulating free DNA (cfDNA) during target therapy could hold significant clinical importance in non-small cell lung cancer (NSCLC). Our aim is to establish if EGFR mutational status change on cfDNA has predictive value that can impact clinical management of NSCLC patients care. METHODS: This study included 30 patients with EGFR-mutated NSCLC. Blood samples were collected at diagnosis (T0) and in 19 patients during therapy (T1). RESULTS: Concordance between T0 and T1 EGFR mutation status for patients evaluable for both samples (n = 19) was 79%, with a sensitivity of 100% (95% CI: 55.5-100.0) and specificity of 60.0% (95% CI: 26.2-86.8). For the patients in oncological therapy with targeted drug and with T1 sample available (n = 18), survival outcomes were evaluated. For both mutation-negative T0 and T1 patients, 12-month progression-free survival (PFS) was 66.7% (95% CI: 27.2-100.0) and 12-month overall survival (OS) was 100% (95% CI: 1.00-1.00); for patients mutated both at T0 and T1, PFS was 22.2% (95% CI: 6.5-75.4%) and OS was 55.6% (95% CI: 20.4-96.1%). CONCLUSION: EGFR mutation status can be assessed using cfDNA for routine purposes and longitudinal assessment of plasma mutation is an easy approach to monitor the therapeutic response or resistance onset.

Mitochondrial transcription factor A (TFAM) shapes metabolic and invasion gene signatures in melanoma.

Mitochondria are central key players in cell metabolism, and mitochondrial DNA (mtDNA) instability has been linked to metabolic changes that contribute to tumorigenesis and to increased expression of pro-tumorigenic genes. Here, we use melanoma cell lines and metastatic melanoma tumors to evaluate the effect of mtDNA alterations and the expression of the mtDNA packaging factor, TFAM, on energetic metabolism and pro-tumorigenic nuclear gene expression changes. We report a positive correlation between mtDNA copy number, glucose consumption, and ATP production in melanoma cell lines. Gene expression analysis reveals a down-regulation of glycolytic enzymes in cell lines and an up-regulation of amino acid metabolism enzymes in melanoma tumors, suggesting that TFAM may shift melanoma fuel utilization from glycolysis towards amino acid metabolism, especially glutamine. Indeed, proliferation assays reveal that TFAM-down melanoma cell lines display a growth arrest in glutamine-free media, emphasizing that these cells rely more on glutamine metabolism than glycolysis. Finally, our data indicate that TFAM correlates to VEGF expression and may contribute to tumorigenesis by triggering a more invasive gene expression signature. Our findings contribute to the understanding of how TFAM affects melanoma cell metabolism, and they provide new insight into the mechanisms by which TFAM and mtDNA copy number influence melanoma tumorigenesis.

Polymorphisms in VEGFA gene affect the antihypertensive responses to enalapril.

PURPOSE: Vascular endothelial growth factor (VEGF) is a potent angiogenic factor that affects blood pressure by promoting vasodilation mediated by nitric oxide. Angiotensin-converting enzyme inhibitors (ACEi) up-regulate the VEGF expression; thus, genetic polymorphisms in the VEGFA gene could affect the antihypertensive responses to these drugs. METHODS: Hypertensive patients (n = 102) were prospectively treated only with the ACEi enalapril for 60 days. We compared the effect of VEGFA polymorphisms on changes in blood pressure after enalapril treatment. In addition, multiple linear regression analysis was carried out to assess the effect of covariates on blood pressure. Genotypes for g.-2578C>A (rs699947), g.-1154G>A (rs1570360), and g.-634G>C (rs2010963) VEGFA polymorphisms were determined, and haplotype frequencies were estimated. RESULTS: Individuals carrying the CA and AA genotypes for the g.-2578C>A polymorphism and the AGG haplotype showed more intense decrease in blood pressure in response to enalapril 20 mg/day. A multiple linear regression analysis showed that the AA genotype for the g.-2578C>A polymorphism and the AGG haplotype are associated with more intense decrease in blood pressure in response to enalapril 20 mg/day, while the CC genotype for the g.-2578C>A polymorphism and the CGG haplotype are associated with the opposite effect. CONCLUSIONS: These findings suggest that polymorphisms in VEGFA gene may affect the antihypertensive responses to enalapril.

Comparative safety profiles of intravitreal bevacizumab, ranibizumab and pegaptanib: the analysis of the WHO database of adverse drug reactions.

PURPOSE: The purpose of this study is to conduct a comparative analysis of the suspected adverse drug reactions (ADRs) associated with intravitreal bevacizumab, ranibizumab and pegaptanib in the WHO database in order to have a real-life information on these drugs, which now is only based on data coming from clinical trials. METHODS: ADR reports for intravitreal use of bevacizumab, ranibizumab and pegaptanib from January 2002 to December 2012 were selected from the WHO-VigiBase. Reporting odds ratio (ROR) with confidence interval of 95 % and p value was calculated. The analysis was performed for drug-reaction pairs. The Medical Dictionary for Regulatory Activities (MedDRA) terminology for ADRs was used. RESULTS: The analysis was performed on 3180 reports corresponding to 7753 drug-reaction pairs. Significant RORs for endophthalmitis and uveitis (1.90, 95 % confidence interval (CI) 1.48-2.43, and 10.62, 6.62-17.05, respectively) were retrieved for bevacizumab, and cerebrovascular accident and myocardial infarction produced significant ROR (1.54, 1.14-2.10 and 1.73, 1.18-2.53, respectively) for ranibizumab. Pegaptanib was significantly associated with visual impairment (1.98, 1.12-3.5, p = 0.02), nausea (3.29, 1.57-6.86, p < 0.001), vomiting (2.91, 1.2-7.07, p = 0.01) and drug hypersensitivity (8.75, 3.1-24.66, p < 0.001). CONCLUSIONS: Our data showed an elevated disproportionality for cardiovascular ADRs in patients treated with ranibizumab and for infective ocular reactions in those treated with bevacizumab. No relevant safety issues were identified for pegaptanib. These findings suggest bevacizumab as a suitable choice for AMD therapy due to its effectiveness similar to that of ranibizumab, its favourable safety profile and for its lower cost.

Benign hepatic nodular lesions after treatment for childhood cancer.

BACKGROUND: Benign nodular hepatic regenerating lesions such as focal nodular hyperplasia (FNH) have been reported as rare complications of the antineoplastic therapy received during infancy. Little is known about the risk factors associated with the onset of these lesions and their diagnostic management. METHODS: We have analyzed a series of benign hepatic nodular lesions occurring in children previously treated for malignant tumors in our institution in a period of 11 years. An extensive description of the imaging presentation of the lesions has been provided to facilitate the differential diagnosis, and a risk factor analysis has been conducted. RESULTS: A total of 14 diagnoses (10 FNH and 4 hemangiomas) of benign nodular hepatic lesions have been found. Hematopoietic stem cell transplantation is the most important statistically independent risk factor associated with the development of these lesions, especially for FNH. No malignant transformation of nodules has been recorded during a median follow-up time of 4 years. CONCLUSIONS: In our experience, FNH is the most frequent benign nodular hepatic lesions occurring after treatment for childhood cancer. Hematopoietic stem cell transplantation is the most important risk factor to be taken in account. After a secure diagnosis of these benign lesions, only a close imaging follow-up is recommended.

eNOS and BDKRB2 genotypes affect the antihypertensive responses to enalapril.

PURPOSE: The antihypertensive effect of angiotensin-converting enzyme inhibitors (ACEi) is attributed partially to increased nitric oxide bioavailability. It is possible that functional polymorphisms in endothelial nitric oxide synthase (eNOS) and bradykinin receptor B2 (BDKRB2) genes may affect the antihypertensive response to enalapril. METHODS: We evaluated 106 hypertensive patients treated only with enalapril for 60 days. The difference between the mean arterial pressure (MAP) before and after the antihypertensive treatment was defined as ΔMAP. If ΔMAP were below or above the median value, the patients were classified as poor responders (PR) or good responders (GR), respectively. eNOS genotypes for the T(-786)C, G894T and 4b/4a polymorphisms were determined and haplotype frequencies were estimated by PHASE and Haplo.stats programs. The C(-58)T and BE1 +9/-9 polymorphisms of BDKRB2 genes and their haplotypes were determined by DNA sequencing. Robust multifactor dimensionality reduction analysis was used to characterize gene-gene interactions. RESULTS: The TC/CC genotypes and the C allele for the eNOS T(-786)C polymorphism were more frequent in GR than in PR. Furthermore, the TT genotype for the BDKRB2 C(-58)T polymorphism was more frequent in PR than GR. No other significant differences in genotypes or haplotypes were found. However, we found significant gene-gene interactions: the CC genotype for the BDKRB2 C(-58)T polymorphism was associated with response to enalapril depending on eNOS T(-786)C genotypes. CONCLUSIONS: These findings suggest that eNOS T(-786)C and BDKRB2 C(-58)T polymorphisms may synergically affect the antihypertensive response to enalapril.

[Vertical transmission of HGV and outcome of the infected babies]. / Trasmissione verticale di HGV e storia naturale dell'infezione nei neonati.

To assess the risk of HGV mother-to-infant transmission and the clinical outcome of infected babies, we investigated 103 mother-infant couples and followed-up the infected children for 4-72 months. Twenty (19.4%) mothers were HGV-RNA positive and transmission occurred in ten (50%) babies; only one child acquired HGV and HCV infection. Maternal factors, such as history of intravenous drug use, HCV-RNA positivity, HIV coinfection, type of delivery and type of feeding were not related to HGV transmission. One HGV infected baby showed a mild hepatitis when he was also infected by Cytomegalovirus. Two babies cleared HGV within the first year of life. The HGV transmission rate is elevated but HGV infection seems to be benign, at least in a short-term follow-up.

Genetic mapping of Y-chromosomal DNA markers in Pacific salmon.

Sex chromosomes in fish provide an intriguing view of how sex-determination mechanisms evolve in vertebrates. Many fish species with single-factor sex-determination systems do not have cytogenetically-distinguishable sex chromosomes, suggesting that few sex-specific sequences or chromosomal rearrangements are present and that sex-chromosome evolution is thus at an early stage. We describe experiments examining the linkage arrangement of a Y-chromosomal GH pseudogene (GH-Y) sequence in four species of salmon (chum, Oncorhynchus keta; pink, O. gorbuscha; coho, O. kisutch; chinook, O. tshawytscha). Phylogenetic analysis indicates that GH-Y arose early in Oncorhynchus evolution, after this genus had diverged from Salmo and Salvelinus. However, GH-Y has not been detected in some Oncorhynchus species (O. nerka, O. mykiss and O. clarki), consistent with this locus being deleted in some lineages. GH-Y is tightly linked genetically to the sex-determination locus on the Y chromosome and, in chinook salmon, to another Y-linked DNA marker OtY1. GH-Y is derived from an ancestral GH2 gene, but this latter functional GH locus is autosomal or pseudoautosomal. YY chinook salmon are viable and fertile, indicating the Y chromosome is not deficient of vital genetic functions present on the X chromosome, consistent with sex chromosomes that are in an early stage of divergence.

[Trend of circulating eosinophils in healthy children and children suffering from infectious diseases. A retrospective study]. / Andamento degli eosinofili circolanti nel bambino sano e nel bambino affetto da malattia infettiva. Studio retrospettivo.

INTRODUCTION: Acute bacterial and viral infections are accompanied by a marked diminution in circulating eosinophils in the blood. This forms part of the host's physiological response to acute infection and was first studied in adults early this century. The aims of this study were to check whether eosinopenia during acute phlogosis is a phenomenon present in pediatric patients, and whether the trend is comparable to the experimental models reported; to describe the trend of circulating eosinophils in the remission process. METHODS: A retrospective study was performed in 34 children hospitalised in the Pediatric Hospital of AUSL 2-Lucca (Italy) for bacterial or viral infective diseases documented by cell culture or presumed diagnosed. Children with the following characteristics were excluded from the study: 1) blood samples collected for hemochrome analysis at times other than normal (7-8 a.m.); 2) cortisone treatment administered up to 5 days prior to blood sample and/or during hospitalisation; 3) positive personal anamnesis for manifest allergic diseases. On admittance (children during acute phase) and at the start of remission, an absolute count of circulating eosinophils was performed in these children using an automatic globule counter. Sixty-six children with non evident infective and/or inflammatory diseases were included in the study as a control group. This group was also selected in the same way as infective subjects. RESULTS: The mean number of circulating eosinophils was 288 (+/- 248) in the control group, 46 (+/- 58) in subjects at the acute phase of infective pathology and 252 (+/- 162) in infective patient during the remission phase. The difference between the two means was statistically significant. This characteristic falling and rising trend of circulating eosinophils was found in 33 of the 34 infective subjects examined. CONCLUSIONS: Eosinophil values found in control subjects are broadly in line with those reported in the literature. Eosinopenia during the course of acute infection and the early rise during remission represent a characteristic phenomenon indicating the body's "normal" response to a non-parasitic infection. Both eosinophil levels, namely in the control group (288/mm3) and in acute-phase subjects (46/mm3), should be regarded as "normal" provided they refer to the appropriate situation. The precocity and precision with which the eosinophil trend follows the phases of the infection underlines the value of the assay of these cells as a reliable parameter for monitoring acute infection. There are also indications that, in an inflammatory situation, the behaviour of circulating eosinophils may provide a practical clinical marker of the predominant lymphocyte pattern (Th1 or Th2), as well as the phase of phlogosis, active or remission.

[Pattern of circulating eosinophils in allergic children suffering from infectious disease]. / Andamento degli eosinofili circolanti nel bambino allergico affetto da malattia infettiva.

BACKGROUND AND AIM: Both bacterial and viral acute infections are accompanied by a typical fall in the number of circulating eosinophils followed, at remission, by a rapid return to starting values. The aim of this study was to check whether this typical pattern can be observed in atopical children. MATERIALS AND METHODS: A retrospective study was performed in a group of atopical subjects (mainly asthmatic) suffering from intercurrent infectious diseases (mainly bronchopneumonia); circulating eosinophils were assayed during the acute phase of disease and during remission. The control group consisted of 21 atopic children without signs or symptoms of infectious diseases. RESULTS: The mean number of circulating eosinophils was 412.6 +/- 199.8 in the control group of allergic children 25.1 +/- 39.7 in the group with acute infectious diseases and 241 +/- 137.5 in the group of children with infectious diseases during remission. The differences between the group of children with infectious diseases and that in remission were statistically significant. The typical pattern of the fall and rise of circulating eosinophils was observed in 9 out of 10 children with acute infections. The cut-off proposed as the discriminating factor between acute phase infection and subjects in remission or without infectious diseases was 100 eosinophils/mm3. CONCLUSIONS: During the course of infective pathologies in allergic subjects there is an abrupt and significant reduction in circulating eosinophils followed, during remission, by a return to levels comparable to those in controls. Eosinophil assay therefore appears to be a useful parameter to monitor acute infection in allergic subjects. In particular, increased eosinophil levels are an early indicator that the infection has been overcome and remission is in progress. The authors repropose the hypothesis that eosinophils play a double role that alternates between activating the allergic phlogosis and eliminating non-allergic phlogosis, depending on the cytokinic context in which this occurs.

[Assessment of lead blood levels in children living in Lucca (Italy) in 1993. The study was conducted as part of the project METOS, in accordance with the decree 496 of June 1982 which adopted the directive CEE No.77/312]. / Livelli ematici di piombo nei bambini della piana di lucca. Rilevazione del 1993. Studio svolto nell'ambito del progetto Metos, in attuazione al D.P.R. 496/giugno 1982 che recepisce la direttiva CEE n degrees 77/312.

OBJECTIVE: Lead poisoning is a major cause of environmental concern in all countries worldwide. Saturnism in children, especially if young and neglected, represents a peculiar phenomenon both in terms of the biology of growing subjects and the epidemiological nature of poisoning. In under five-year-olds, in addition to lead levels in the atmosphere, it is equally important to evaluate the presence of contaminated dust in the house, hand-to-mouth activities and the level of care provided by parents. It has been demonstrated that, in the presence of equivalent environmental lead levels, dust removal from the house and prevention of hand-to-mouth activities can successfully reduce lead blood levels (PbB) in children. AIM: The aim of this study was to evaluate the risk of chronic lead poisoning in children attending Local Health Unit 6 in Piana di Lucca and to assess the need for a possible health education campaign aimed at eliminating the specific pediatric risk for lead poisoning. MATERIALS AND METHODS: Assay of lead blood levels in 172 children from 0 to 14 years old consecutively attending the Emergency Ward at Campo di Marte Hospital in Lucca for reasons not relating to lead poisoning. RESULTS: Mean lead blood levels in children were 57.2 +/- 30.2 mg/l (mean 50 mg/l) without significant differences between the various age brackets (0-5, 6-10, 11-14 years old). Only one child presented pathological PbB levels (280 mg/l when first measured; 360 mg/l a few months later with normal values of erythrocytic Zn-protoporphyrin, 24-h urinary lead excretion and 24-h urinary delta-aminolevulinic acid). An epidemiological study is now being made of the causes. The general population within the same area presented mean PbB levels of 70.4 +/- 36.9 mg/l (mean 60 mg/l), measured in a total of 471 subjects (adults and children). DISCUSSION AND CONCLUSIONS: The PbB levels currently found in children resident in Lucca confirm a low-risk environmental situation. Preventive measures specifically aimed at children do not therefore appear to be justified at present.