Occurrence and transformation of illicit drugs in wastewater treatment plants.

Illicit drugs (IDs) and their metabolites have been recently recognized as a new group of water emerging contaminants (ECs) with potent psychoactive properties and unknown effects to the aquatic environment (Pal et al., 2013). IDs are excreted via urine and feces and arrive at wastewater treatment plants (WWTPs) where can reach ppb levels (Castiglioni et al., 2006). Over the past few years, it has been demonstrated that conventional biological processes in WWTPs are not or scarcely able to remove IDs. Thus, they are discharged into water bodies through the treated effluent (Postigo et al., 2011). Therefore, monitoring the IDs concentration in WWTPs can have a twofold advantage: i. increase knowledge on the amount of IDs discharged in the environment and estimate their effect; ii. estimating indirectly the community level consumption (Senta et al., 2014). The objective of this paper is to provide a comprehensive analysis of the occurrence and behaviour of illicit drugs and their metabolites in two Sicilian WWTPs. Specifically, two WWTPs (namely, WWTP-1 and WWTP-2) located at the north-western Sicilian coast have been monitored for 5 months (one sampling per week). The two WWTPs have a conventional scheme and mainly differ for their potentiality. Indeed, the average daily flow expressed as m3d-1 for WWTP-1 and WWTP-2 was equal to 153,600 and 19,704, respectively. Samples were analyzed for total suspended solids (TSS), illicit drugs and their metabolites (metham-phetamine; COC = cocaine; MDMA = 3,4-methylenedioxymethamphetamine; METH = methadone; EDDP = 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine; MDA = 3,4-methylenedioxy amphetamine; MDEA = 3,4-methylenedioxy ethylamphetamine; THC-COOH = 11-nor-9-carboxy-\u3949-tetrahydrocannabinol; BEG= Benzoylecgonine). In order to provide a fast and sensitive approach to quantify IDs, an automated online sample preparation method has been developed. The method uses a Thermo Scientific Transcend TLX-1 system powered by TurboFlowTM technology coupled with a TSQ Quantiva Triple Quadrupole Mass Spectrometer. Specifically, THC-COOH has been extracted from 75 \ub5L of pre-filtered water (using 7 and 0.4 \ub5m paper filters) by an online sample extraction method and quantified using an isotopic dilution approach between 30 and 2000 ng L-1

EGFR CELL EXPRESSION IN BLADDER WASHINGS AS A RISK MARKER TOOL IN NON MUSCLE-INVASIVE BLADDER CANCER. PRELIMINARY EXPERIENCE

INTRODUCTION AND OBJECTIVES: Up to day, EGFR expression has been determined mainly in tissue specimens of muscleinvasive bladder cancer and its overexpression has been associated with worse prognosis and shorter survival. Urothelial EGFR status after NMIBC transurethral resection (TUR) could indicate the risk of recurrence and progression. We investigated the feasibility of EGFR measurement in bladder washings of patients undergoing intravesical adjuvant therapy for NMIBC and its usefulness in identifying risk subgroups. METHODS: Our prospective study included patients after TUR of NMIBC and healthy controls. A cellular pellet was obtained from bladder washing, and RNA extraction performed by miRNeasy Mini Kit (Qiagen). Good quality of RNA was checked. The cDNA obtained from RNA was used to perform a gene expression analysis by a Real Time PCR, according to the method of the comparative quantification (DDCt) with an endogenous control (Cyclophilin). Every reaction was set in triplicate as a guarantee of quality. Patients were grouped for EAU risk class and maintained in follow-up. The EGFR expressions were statistically analyzed according to EAU risk groups and to patients0 outcome. EGFR gene expression values were expressed in FOLDs of change compared to healthy controls (EGFR¼1). RESULTS: Fifty-eight patients and 21 healthy age-matched controls were entered. An adequate cellular pellet was obtained in 50 patients (86.2%) showing a median EGFR expression of 2.0 folds (IQR 0.6-4.3, p¼0.0004). After TUR and adjuvant intravesical therapy, 22 (55%) out of 40 high-risk patients, showed EGFR decrease to 1.3 folds (IQR 0.9-1.5), while 18 (45%) showed elevated EGFR, median 4.7 (IQR 4.1-11.6). At 25 months median follow-up (IQR 19.0-34.8), 20 (40%) patients recurred and 6 (12%) progressed. Among patients with or without EGFR gene increase, 9 (22.5%) and 5 (12.5%) recurred and 5 (12.5%) and 1 (2.5%) progressed, respectively. CONCLUSIONS: In our experience EGFR expression measurement was feasible in more than 85% of patients and resulted related to EAU risk classes for recurrence and progression, showing different behavior during intravesical therapy. It was possible to identify a subgroup of high risk patients overexpressing EGFR in spite of intravesical adjuvant therapy. EGFR evaluation in bladder washing could represent a repeatable and useful tool to identify a subgroup of patients at risk for progression unresponsive to intravesical adjuvant therapy and candidate to early radical cystectom

TM6SF2 rs58542926 is not associated with steatosis and fibrosis in largecohort of patients with genotype 1 chronic hepatitis C

Background & Aims: We tested the putative association of the rs58542926 variant of TM6SF2, a recently described genetic determinant of nonalcoholic fatty liver disease, with steatosis and fibrosis in genotype 1(G1) chronic hepatitis C(CHC) patients. Methods: A total of 694 consecutively biopsied Caucasian G1 CHC patients were genotyped for TM6SF2 rs58542926, IL28B rs12979860 and PNPLA3 rs738409. Steatosis was classified as absent (<5%), mild-moderate(5-29%) and severe( 6530%), Fibrosis was considered severe if=F3-F4. Results: Carriers of TM6SF2 rs58542926 (6.3% of patients) exhibited lower serum levels of cholesterol (P=0.04) and triglycerides (P=0.01), but a similar distribution of steatosis severity (P=0.63), compared to noncarriers. Prevalence and severity of steatosis were reduced in IL28B C allele carriers (P=0.005) and elevated in PNPLA3G allele carriers (P<0.001). After adjustment for age, gender, body mass index and homoeostasis model assessment score, steatosis severity was independently associated with IL28B rs12979860 (odds ratio [OR] 0.69, 95% confidence interval [CI] 0.55-0.86, P=0.001) and PNPLA3 rs738409 (OR 1.84, 95% CI 1.46-2.83, P<0.001), but not TM6SF2 rs58542926 (OR 1.48, 95% CI 0.82-2.69, P=0.19). Variants of TM6SF2 (30.9% vs. 25%, P=0.40), IL28B and PNPLA3 were not directly associated with fibrosis severity, although variants of IL28B and PNPLA3 promoted steatosis (OR 1.36, 95% CI 1.06-1.75, P=0.01) that in turn is associated with severe fibrosis. Conclusions: In G1 CHC patients, TM6SF2 rs58542926 does not affect the histological severity of liver damage. However, IL28B rs12979860 and PNPLA3 rs738409 modify steatosis

Respiratory Syncytial Virus: New Challenges for Molecular Epidemiology Surveillance and Vaccination Strategy in Patients with ILI/SARI.

Abstract: Several respiratory pathogens are responsible for influenza-like illness (ILI) and severe respiratory infections (SARI), among which human respiratory syncytial virus (hRSV) represents one of the most common aetiologies. We analysed the hRSV prevalence among subjects with ILI or SARI during the five influenza seasons before the emergence of SARS-CoV-2 epidemic in Sicily (Italy). Respiratory specimens from ILI outpatients and SARI inpatients were collected in the framework of the Italian Network for the Influenza Surveillance and molecularly tested for hRSV-A and hRSV-B. Overall, 8.1% of patients resulted positive for hRSV. Prevalence peaked in the age-groups &lt;5 years old (range: 17.6–19.1%) and ≥50 years old (range: 4.8–5.1%). While the two subgroups co-circulated throughout the study period, hRSV-B was slightly predominant over hRSV-A, except for the season 2019–2020 when hRSV-A strongly prevailed (82.9%). In the community setting, the distribution of hRSV subgroups was balanced (47.8% vs. 49.7% for hRSV-A and hRSV-B, respectively), while most infections identified in the hospital setting were caused by hRSV-B (69.5%); also, this latter one was more represented among hRSV cases with underlying diseases, as well as among those who developed a respiratory complication. The molecular surveillance of hRSV infections may provide a valuable insight into the epidemiological features of ILI/SARI. Our findings add new evidence to the existing knowledge on viral aetiology of ILI and SARI in support of public health strategies and may help to define high-risk categories that could benefit from currently available and future vaccines

Comparison of Histochemical Stainings in Evaluation of Liver Fibrosis and Correlation with Transient Elastography in Chronic Hepatitis

Background and Aim. The best staining to evaluate liver fibrosis in liver hepatitis is still a debated topic. This study aimed to compare Masson's trichrome (MT), Sirius Red (SR), and orcein stainings in evaluating liver fibrosis in chronic HCV hepatitis (CHC) with semiquantitative and quantitative methods (Collagen Proportionate Area (CPA) by Digital Image Analysis (DIA)) and correlate them with transient elastography (TE). Methods. Liver stiffness evaluation of 111 consecutive patients with CHC was performed by TE. Semiquantitative staging by Metavir score system and CPA by DIA were assessed on liver biopsy stained with MT, SR, and orcein. Results. MT, SR, and orcein staining showed concordant results in 89.6% of cases in staging CHC, without significant difference in both semiquantitative and quantitative evaluations of fibrosis. TE values were concordant with orcein levels in 86.5% of the cases and with MT/RS in 77.5% (P < 0.001). No significant correlation between the grade of necroinflammatory activity and TE values was found. Conclusion. In CHC, SR/MT and orcein stainings are almost concordant and when discordant, orcein staining is better related to TE values than MT/RS. This suggests that elastic fibers play a more important role than reticular or collagenous ones in determining stiffness values in CHC

β-amyloid wall deposit of temporal artery in subjects with spontaneous intracerebral haemorrhage.

Background: Cerebral Amyloid Angiopathy has been indicated as an important cause of spontaneous non-hypertensive intracerebral haemorrhage (ICH). Aims: to analyze the presence of β-amyloid deposit in the temporal artery of consecutive patients with ICH in comparison to control subjects and its relation to APO-E haplotype frequency. Methods: We enrolled consecutive patients admitted to Neurosurgery Ward of University Hospital "P. Giaccone" of Palermo with a diagnosis of spontaneous non hypertensive ICH and as control 12 subjects without brain haemorrhage. Biopsy of superficial temporal artery has been performed and β-amyloid deposit was quantified. Results: Among 25 subjects with ICH, 10 (40%) had APOE epsilon 2 allele and among these subjects 7 (70%) showed amyloid accumulation on temporal artery specimens, 8 (32%) subjects had APOE epsilon 3 allele and among these subjects only 2 (25%) showed amyloid accumulation on temporal artery specimens, whereas 7 (28%) had APOE epsilon 4 allele and of these, 7 (100%) showed amyloid accumulation on temporal artery specimens. At multivariable logistic regression analysis for the presence of amyloid, predictive factors for the presence of amyloid in temporal artery biopsies were: age, hypertension, intralobar site of haemorrhage, APOE epsilon 2 and APOE epsilon 4 alleles. Discussion: Our findings of a higher frequency of amyloid deposition in temporal artery specimens in subjects with spontaneous intracerebral haemorrhage indicate a possible role of temporal artery as a possible diagnostic site of biopsy in subjects at high risk to develop intracranial haemorrhage related to Cerebral Amyloid Angiopath