EFFICIENT PROPAGATION OF ARCHETYPE JC POLYOMAVIRUS IN COS-7 CELLS: EVALUATION OF REARRANGEMENTS WITHIN NCCR STRUCTURAL ORGANIZATION DURING TRANSFECTION.

John Cunningham virus (JCPyV) is an ubiqui-tous human pathogen that causes disease in immunocom-promised patients. The JCPyV genome is composed of an early region and a late region, which are physically sepa-rated by the non-coding control region (NCCR). The DNA sequence of the NCCR distinguishes two forms of JCPyV, the designated archetype and the prototype, which resulted from a rearrangement of the archetype sequence. To date, the cell culture systems for propagating JCPyV archetype have been very limited in their availability and robust-ness. Prior to this study, it was demonstrated that JCPyV archetype DNA replicates in COS-7 simian kidney cells expressing SV40 TAg and COS-7 cells expressing HIV-1 Tat. Based on these observations, the present study was conducted to reproduce an in vitro model in COS-7 cells transfected with the JCPyV archetype strain in order to study JCPyV DNA replication and analyze NCCR rear-rangements during the viral life cycle. The efficiency of JCPyV replication was evaluated by quantitative PCR (Q-PCR) and by hemagglutination (HA) assay after trans-fection. In parallel, sequence analysis of JCPyV NCCR was performed. JCPyV efficiently replicated in kidney-derived COS-7 cells, as demonstrated by a progressive increase in viral load and virion particle production after transfection. The archetypal structure of NCCR was maintained during the viral cycle, but two characteristic point mutations were detected 28 days after transfection. This model is a useful tool for analyzing NCCR rearrangements during in vitroreplication in cells that are sites of viral persistence, such as tubular epithelial cells of the kidne

Small bowel infarction by Aspergillus.

Primary gut involvement by Aspergillus is a rare and often fatal complication of intensive antileukemic therapy. We describe the case of an adult patient affected by acute leukemia who developed a small bowel fungal thromboembolism without radiographic evidence of lung involvement during the post-induction aplastic phase. The diagnosis was made histologically at laparotomy performed for small bowel perforation. The patient died a week later in spite of amphotericin-B treatment and neutrophil recovery. Anti- Aspergillus prophylaxis and early introduction of amphotericin-B in the treatment of febrile neutropenia is probably advisable in all cases of AML

Tinnitus revival during COVID‑19 lockdown: how to deal with it?

To the Editor, The novel Coronavirus Disease, officially designated as COVID-19 by the WHO, is a serious issue for public health. To contain the COVID-19, the Italian Government stated on March 9th 2020 the prohibition of any movement throughout the national area unless for work/health reasons and the obligation to remain as much as possible inside one’s own home. With the start of the so-called “Phase Two” on May 4th 2020, circulation within the same region was allowed again, due to the progressive slowdown of the outbreak. Therefore, since lockdown measures were relaxed and access to the emergency room or ENT clinic became less worrying for patients, specialists of Otolaryngology Units in Bari (Italy) observed an increase in the amount of subjects complaining of the revival of intense tinnitus. We attempt in this letter to focus on patients affected by chronic subjective tinnitus, that already had a diagnosis and self-stabilized without a massive treatment. Research studies have reported tinnitus wide impact on quality of life of subjects experiencing it, involving their emotional state, concentration and sleep quality; at this regard, Tinnitus Handicap Inventory (THI) is a self-reported tool regularly used to quantify the grade of perceived handicap as slight (0–16), mild (18–36), moderate (38–56), severe (58–76) or catastrophic (78–100) on the basis of 25 questions [1]. During the past 2 weeks, we have collected data from 16 patients among our population of chronic sufferers: THI observed was moderate in 62.5% and severe in 18.75% of cases, catastrophic in 12.5% and mild in 6.25% of subjects. Interestingly, the grade of handicap resulted increased by one-level in 12 out of 16 patients (75%); in particular, THI shifted from mild to moderate in 9 patients and from moderate to severe in 3 patients. As shown in the literature, tinnitus generation, maintenance and recrudescence are still debated. A cortical reorganization secondary to sensory deprivation has been proposed as one on the most frequent cause of tinnitus [2]. The avoidance of silence and acoustic masking have been proposed as effective measures to overcome sensory deprivation and increase masking of the symptom [3]. It is reasonable to think that, during the lockdown, the absence of environmental masking sounds from everyday life may have enhanced the tinnitus perception. Furthermore, proneness to worry and incoming stress during pandemic could be included as further potential risk factors for tinnitus worsening. As proficiently reviewed in a recent work [4], some internet/smartphone-based applications provide in tinnitus patients adequate counseling and interactive information together with sound therapy. As brain networks implicated in adaptive responses to sound stimuli and to worry are shared in many cases, an early decrease of anxiety status may release neural resources crucial for tinnitus habituation/distress perception [5]. In general, interactive platforms have been widely implemented during lockdown period due to the forced lack of real personal and working relationships; since smart-working seems to be successful for future plans, the development of smart applications and mobile services in the health care field may be promising in terms of cost-effectiveness, tolerability and simplicity of use

Epidemiology of Herpes Simplex Virus Infection in Pregnancy: A Pilot Study:

Herpes simplex virus (HSV) infection is one of the most common sexually transmitted viral diseases worldwide. HSV type 2 causes most genital herpes and HSV type 1 is usually transmitted via non-sexual contacts. We studied 109 pregnant women between January 2007 and December 2008, in relation to their age, condom use, number of sexual partners, age at first intercourse, parity and smoking habits. The aim of this study is to evaluate the prevalence of HSV cervical infection and HSV co-infection with other genital microorganisms associated with poor neonatal outcome. Our results show that of the 109 outpatients enrolled, 30% were HSV1 and/or HSV2 positive, of whom 30% were infected with both HSV1 and HSV2, 18% were infected with HSV1 alone and 52% with HSV2 alone. A significant association between HSV1 and HSV2 infection was found, and the prevalence of HSV2 infection in women infected with HSV1 was 63%. The prevalence of HSV1/2 varied in the presence of other vaginal microorganisms but a statistical significant association was not found. This pilot study is probably too small to obtain statistically significant results. Nevertheless, using these observed results, we calculated that about 530 patients with comparable features should be enrolled to detect an increase of 50% in HSV infection due to the presence of other genital infections and potential risk factors

Age-related changes in bile acid synthesis and hepatic nuclear receptor expression

BACKGROUND:Recent data highlighted the role of nuclear receptors in the transcriptional regulation of the limiting enzyme of bile acid synthesis, cholesterol 7alpha-hydroxylase, in cellular and animal models. This study was designed to analyze the effects of age on cholesterol 7alpha-hydroxylase and related nuclear receptor expression in human livers.DESIGN:Surgical liver biopsies were obtained in 23 patients requiring operation on the gastrointestinal tract. mRNA levels of cholesterol 7alpha-hydroxylase and related nuclear receptors and co-activators were assayed by quantitative real-time RT-PCR. Serum levels of 7alpha-hydroxy-4-cholesten-3-one, a marker of bile acid synthesis, were assayed by gas-liquid chromatography:mass spectrometry.RESULTS:Ageing was inversely correlated with serum 7alpha-hydroxy-4-cholesten-3-one and with cholesterol 7alpha-hydroxylase mRNA levels (r = -0.44 and r = -0.45 on a semi-log scale, respectively, P < 0.05). Among different nuclear factors, cholesterol 7alpha-hydroxylase mRNA best correlated with hepatocyte nuclear factor-4 (r = 0.55 on a log scale, P < 0.05); hepatocyte nuclear factor-4 levels were also inversely correlated with age (r = -0.64 on a semi-log scale, P < 0.05). Age was inversely correlated with serum insulin-like growth factor-I levels, which were directly correlated with hepatocyte nuclear factor-4 and cholesterol 7alpha-hydroxylase expression. No suppressive effect of short heterodimer partner expression on cholesterol 7alpha-hydroxylase was observed.CONCLUSIONS:Ageing associates with reduced bile acid synthesis, possibly related to decreased hepatic expression of hepatocyte nuclear factor-4 and consequently of cholesterol 7alpha-hydroxylase. Age-related modifications of the growth hormone/insulin-like growth factor axis might play a role. These findings may help to elucidate the pathophysiology of age-related modifications of cholesterol metabolism

Natalizumab affects T-cell phenotype in multiple sclerosis: implications for JCV reactivation

The anti-CD49d monoclonal antibody natalizumab is currently an effective therapy against the relapsing-remitting form of multiple sclerosis (RRMS). Natalizumab therapeutic efficacy is limited by the reactivation of the John Cunningham polyomavirus (JCV) and development of progressive multifocal leukoencephalopathy (PML). To correlate natalizumab-induced phenotypic modifications of peripheral blood T-lymphocytes with JCV reactivation, JCV-specific antibodies (serum), JCV-DNA (blood and urine), CD49d expression and relative abundance of peripheral blood T-lymphocyte subsets were longitudinally assessed in 26 natalizumab-treated RRMS patients. Statistical analyses were performed using GraphPad Prism and R. Natalizumab treatment reduced CD49d expression on memory and effector subsets of peripheral blood T-lymphocytes. Moreover, accumulation of peripheral blood CD8+ memory and effector cells was observed after 12 and 24 months of treatment. CD4+ and CD8+ T-lymphocyte immune-activation was increased after 24 months of treatment. Higher percentages of CD8+ effectors were observed in subjects with detectable JCV-DNA. Natalizumab reduces CD49d expression on CD8+ T-lymphocyte memory and effector subsets, limiting their migration to the central nervous system and determining their accumulation in peripheral blood. Impairment of central nervous system immune surveillance and reactivation of latent JCV, can explain the increased risk of PML development in natalizumab-treated RRMS subjects

Quartz fiber calorimetry

The fundamentals of a new electromagnetic and hadronic sampling calorimetry based on the detection of Cherenkov light generated in quartz optical fibers are presented. Optical fibers transport light only in a selected angular range which results in a non-obvious and absolutely unique characteristic for this new technique: showers of very narrow visible energy. In addition, the technique is characterized by radiation resistance measured in Gigarads and nanosecond signal duration. Combined, these properties make quartz fiber calorimetry a very promising technique for high intensity heavy ion experiments and for the high pseudorapidity regions of high intensity collider experiments. The results of beam tests and simulations are used to illustrate the basic properties and peculiar characteristics of this recent development

Prospects for K+→π+ννˉK^+ \to \pi^+ \nu \bar{ \nu } at CERN in NA62

The NA62 experiment will begin taking data in 2015. Its primary purpose is a 10% measurement of the branching ratio of the ultrarare kaon decay $K^+ \to \pi^+ \nu \bar{ \nu }$, using the decay in flight of kaons in an unseparated beam with momentum 75 GeV/c.The detector and analysis technique are described here.Comment: 8 pages for proceedings of 50 Years of CP

A precision measurement of direct CP violation in the decay of neutral kaons into two pions

The direct CP violation parameter Re(epsilon'/epsilon) has been measured from the decay rates of neutral kaons into two pions using the NA48 detector at the CERN SPS. The 2001 running period was devoted to collecting additional data under varied conditions compared to earlier years (1997-99). The new data yield the result: Re(epsilon'/epsilon) = (13.7 +/- 3.1) times 10^{-4}. Combining this result with that published from the 1997, 98 and 99 data, an overall value of Re(epsilon'/epsilon) = (14.7 +/- 2.2) times 10^{-4} is obtained from the NA48 experiment.Comment: 19 pages, 5 figures, to be published in Physics Letters

Measurement of the branching ratios of the decays Xi0 --> Sigma+ e- nubar and anti-Xi0 --> anti-Sigma+ e+ nu

From 56 days of data taking in 2002, the NA48/1 experiment observed 6316 Xi0 --> Sigma+ e- nubar candidates (with the subsequent Sigma+ --> p pi0 decay) and 555 anti-Xi0 --> anti-Sigma+ e+ nu candidates with background contamination of 215+-44 and 136+-8 events, respectively. From these samples, the branching ratios BR(Xi0 --> Sigma+ e- nubar)= (2.51+-0.03stat+-0.09syst)E(-4) and BR(anti-Xi0 --> anti-Sigma+ e+ nu)= (2.55+-0.14stat+-0.10syst)E(-4) were measured allowing the determination of the CKM matrix element |Vus| = 0.209+0.023-0.028. Using the Particle Data Group average for |Vus| obtained in semileptonic kaon decays, we measured the ratio g1/f1 = 1.20+-0.05 of the axial-vector to vector form factors.Comment: 16 pages, 11 figures Submitted to Phys.Lett.