IMMUNOMODULATORY EFFECTS OF NITAZOXANIDE AND RELATED MOLECULES

Background: Nitazoxanide (Alinia\uae, NTZ) and its active circulating metabolite tizoxanide (TIZ) belong to a new class of anti-infective agents active against parasites, anaerobic bacteria, and viruses. Nowadays, NTZ is licensed in the United States for the treatment of diarrhea caused by Cryptosporidium parvum and Giardia lamblia in adults and children older than twelve months of age. The amplitude of the spectrum of pathogens targeted by NTZ and new-generation non-nitro thiazolides, makes it very unlikely that the action of these compounds is mediated by a pathogen-specific mechanism(s), suggesting instead that thiazolides act as immunomodulants. To date, the potential effect of these compounds on immune responses has nevertheless not been analysed. In particular, because innate immunity and type 1 interferons are pivotal in early and effective antiviral immune responses that are not antigen-restricted, it is plausible to hypothesize that thiazolides could potentiate this arm of the immune system. To verify this possibility, we performed extensive in vitro analyses on the immunomodulatory effects of TIZ and the second generation non-nitro thiazolide RM4848 using two different models of viral infection: Influenza and HIV-1. Methods: Peripheral blood mononuclear cells (PBMCs) from 20 healthy donors were stimulated with influenza virus antigen (FLU) or infected with HIV-1BaL strain and cultured in presence/absence of TIZ or RM4848. Thiazolide effects on innate immunity were examined by evaluating TLRs expression on monocytes, IFN-secretion by dendritic cells, cytokine and chemokine production, mRNA expression of multiple genes involved in TLR, type I IFN and in cholesterol metabolism pathways. Results: Thiazolides are associated with strong immunomodulatory effects. Notably, these compounds, both in FLU-stimulated and in HIV-1-infected cells, significantly increase: 1) TLR-expression on monocytes, 2) IFN production, 3) chemokine and cytokine production, 4) mRNA expression of different genes operating in the TLR and type I IFN pathways, 5) genes involved in cholesterol metabolism and efflux. Conclusions: Data herein show that thiazolides are potent type I IFN inducers, triggering a selective activation of several IFN-stimulated gene (ISG) pathway. Thus, increased expression of innate antiviral factors and the different modulation of genes involved in cholesterol metabolism and efflux suggest a new mechanism of action mediated by thiazolides

A Reconfigurable Impedance Matching Network Employing RF-MEMS Switches

We propose the design of a reconfigurable impedance matching network for the lower RF frequency band, based on a developed RF-MEMS technology. The circuit is composed of RF-MEMS ohmic relays, metal-insulator-metal (MIM) capacitors and suspended spiral inductors, all integrated on a high resistivity Silicon substrate. The presented circuit is well-suited for all applications requiring adaptive impedance matching between two in principle unknown cascaded RF-circuits. The fabrication and testing of a monolithic integrated prototype in RF-MEMS technology from ITC-irst is currently underway.Comment: Submitted on behalf of EDA Publishing Association (http://irevues.inist.fr/EDA-Publishing

Evaluation of Toxicant-Associated Fatty Liver Disease and Liver Neoplastic Progress in Sprague-Dawley Rats Treated with Low Doses of Aflatoxin B1 Alone or in Combination with Extremely Low Frequency Electromagnetic Fields

The term toxicant-associated fatty liver disease (TAFLD) has been proposed to describe fatty liver diseases connected to toxicants other than alcohol. Aflatoxins are mycotoxins commonly found as contaminants in foods and feeds, which are known liver toxicants and potential candidates as potential causes of TAFLD. Aflatoxin B1 (AFB1) was administered at low doses to Sprague-Daw-ley (SD) rats, alone or in combination with S-50 Hz an extremely low frequency electromagnetic field (ELFEMF), to study the evolution of TAFLD, preneoplastic and neoplastic lesions of the liver and the potential enhancing effect of lifespan exposure to ELFEMF. Steatosis, inflammation and foci of different types were significantly increased in both aflatoxin-treated males and females, which is consistent with a pattern of TAFLD. A significant increase in adenomas, cystic dilation of biliary ducts, hepatocellular hyperplasia and hypertrophy and oval cell hyperplasia were also observed in treated females only. The administration of low doses of AFB1 caused TAFLD in SD rats, inducing liver lesions encompassing fatty infiltration, foci of different types and adenomas. Furthermore, the pattern of change observed in preneoplastic liver lesions often included liver steatosis and steato-hepatitis (TASH). ELFEMF did not result in any enhancing or toxic effect in the liver of SD rats

Aflatoxin B1 DNA-Adducts in Hepatocellular Carcinoma from a Low Exposure Area

Aflatoxin B1 (AFB1) is a class 1 carcinogen with an ascertained role in the development of hepatocellular carcinoma (HCC) in high exposure areas. Instead, this study aimed to assay whether chronic/intermittent, low-dose AFB1 consumption might occur in low-exposure geographical areas, ultimately accumulating in the liver and possibly contributing to liver cancer. AFB1-DNA adducts were assayed by immunostaining in liver tissues from three Italian series of twenty cirrhosis without HCC, 131 HCC, and 45 cholangiocarcinoma, and in an AFB1-induced HCC rat model. CD68, TP53 immunostaining, and TP53 RFLP analysis of R249S transversion were used to characterize cell popula-tions displaying AFB1-DNA adducts. Twenty-five HCCs displayed AFB1-adducts both in neoplastic hepatocytes and in cells infiltrating the tumor and non-tumor tissues. Nuclear immunostaining was observed in a few cases, while most cases showed cytoplasmic immunostaining, especially in CD68-positive tumor-infiltrating cells, suggestive for phagocytosis of dead hepatocytes. Similar patterns were observed in AFB1-induced rat HCC, though with higher intensity. Cholangiocarcinoma and cirrhosis without HCC did not displayAFB1-adducts, except for one case. Despite not providing a causal relationship with HCC, these findings still suggest paying attention to detection and control measures for aflatoxins to ensure food safety in low exposure areas

Novel TCAD Approach for the Investigation of Charge Transport in Thick Amorphous SiO2 Insulators

A TCAD approach for the investigation of charge transport in thick amorphous silicon dioxide is presented for the first time. Thick oxides are investigated representing the best candidates for integrated galvanic insulators in future power applications. The large electric fields, such devices experience and the preexisting defects in the amorphous material, give rise to a leakage current, which leads to degradation and failure. Hence, it is crucial to have a complete understanding of the main physical mechanisms responsible for the charge transport in amorphous silicon oxide. For this reason, metal-insulator-metal structures have been experimentally characterized at different high-field stress conditions and a TCAD approach has been implemented in order to gain insight into the microscopic physical mechanisms responsible for the leakage current. In particular, the role of charge injection at contacts and charge build-up due to trapping-detrapping mechanisms in the bulk of the oxide layer has been investigated and modeled to the purpose of understanding the oxide behavior under dc- and ac-stress conditions. Numerical simulations have been compared against experiments to quantitatively validate the proposed approach

Plasma and PBMC miRNA profile in sexually HIV-1 exposed seronegative individuals

Background: MicroRNAs (miRNAs) are small 20- to 24-nt non-coding RNAs involved in the post-transcriptional regulation of gene expression which play important defensive roles in several viral infections. Global expression profiles of cellular miRNAs have identified alterations of specific miRNAs post-HIV-1 infection both in vitro and in different patient cohorts suggesting potential roles for miRNA in pathogenesis and disease progression. We therefore decided to verify if natural resistance to HIV-1 infection observed in seronegative individuals repeatedly exposed to HIV-1 (HESN) through unprotected sexual intercourse could be secondary to a different expression of their miRNA profile. Methods: Expression levels of 25 miRNAs selected according to their proven anti-HIV-1 properties were analyzed in plasma, basal PBMC and in in vitro HIV-1 infected macrophages isolated from 30 HESN, 30 HIV seropositive subjects (HIV + ) and 30 healthy controls (HC).Results: In plasma the expression of mir-155, mir-382, mir-28 and mir-198 was significantly augmented in both HIV + and HESN compared to HC probably as a consequence of viral exposure. Conversely the expression of mir-223 and mir-150 in plasma was significantly increased only in HESN and this result was also confirmed in basal PBMC suggesting a protective effect for these miRNAs in resistance to HIV-1 infection. Furthermore, the expression of mir-150 was significantly increased in HESN macrophages following HIV-1 infection. Conclusions: mir-223 and mir-150 can target the 3\ua2UTR of HIV-1 transcripts, and they have already been identified as anti-HIV-1 miRNAs. The higher expression of these miRNA in HESN samples could therefore represent a key protection mechanism against HIV infection

Thiazolides elicit anti-viral innate immunity and reduce HIV replication

Nitazoxanide (Alinia (R), NTZ) and tizoxanide (TIZ), its active circulating metabolite, belong to a class of agents known as thiazolides (TZD) endowed with broad anti-infective activities. TIZ and RM-4848, the active metabolite of RM-5038, were shown to stimulate innate immunity in vitro. Because natural resistance to HIV-1 infection in HIV-exposed seronegative (HESN) individuals is suggested to be associated with strong innate immune responses, we verified whether TIZ and RM-4848 could reduce the in vitro infectiousness of HIV-1. Peripheral blood mononuclear cells (PBMCs) from 20 healthy donors were infected in vitro with HIV-1(BaL) in the presence/absence of TIZ or RM4848. HIV-1 p24 were measured at different timepoints. The immunomodulatory abilities of TZD were evaluated by the expression of type I IFN pathway genes and the production of cytokines and chemokines. TZD drastically inhibited in vitro HIV-1 replication (>87%). This was associated with the activation of innate immune responses and with the up-regulation of several interferon-stimulated genes (ISGs), including those involved in cholesterol pathway, particularly the cholesterol-25 hydroxylase (CH25H). TZD inhibition of HIV-1 replication in vitro could be due to their ability to stimulate potent and multifaceted antiviral immune responses. These data warrant the exploration of TZD as preventive/therapeutic agent in HIV infection

Use of contrast-enhanced ultrasonography in chronic pathologic canine testes

Contrast-enhanced ultrasound with sulphur hexafluoride microbubbles was performed in seven healthy dogs without a history of reproductive pathology and with histologically confirmed normal testes and in 42 dogs with chronic scrotal anomalies. All dogs underwent orchiectomy and histological examination. Enhancement patterns and perfusion parameters (peak intensity and regional blood flow) of testes of healthy dogs and testes with chronic lesions were compared. Fourteen non-pathologic and 60 pathologic testes were considered. Forty testes were neoplastic (24 interstitial cell tumours, 9 seminomas, 7 Sertoli cell tumours), 20 were non-neoplastic (16 testicular degenerations, 2 chronic orchitis, 1 testicular atrophy, 1 interstitial cell hyperplasia). In healthy dogs, the contrast medium flow had a rapid homogeneous wash-in and wash-out, with a short peak phase. With contrast ultrasound, testes that were inhomogeneous with a hyperenhancing pattern were associated with neoplasia (sensitivity: 87.5%, specificity: 100%). Lesions with persistent inner vessels and a hypoto-isoechoic background were significantly associated with seminomas (sensitivity: 77.8%, specificity: 100%). Testes with non-neoplastic lesions were characterized by a scant/moderate homogeneous enhancement. Perfusion parameters were higher in neoplastic lesions. Contrast ultrasound was a feasible diagnostic tool in the assessment of testicular lesions, with hyperenhancement being an important feature in the diagnosis of malignancy

A regulatory polymorphism in HAVCR2 modulates susceptibility to HIV-1 infection

The HAVCR2 gene encodes TIM-3, an immunoglobulin superfamily member expressed by exhausted CD8+ T cells during chronic viral infection. We investigated whether genetic variation at HAVCR2 modulates the susceptibility to HIV-1 acquisition; specifically we focused on a 3\u2032 UTR variant (rs4704846, A/G) that represents a natural selection target. We genotyped rs4704846 in three independent cohorts of HIV-1 exposed seronegative (HESN) individuals with different geographic origin (Italy and Spain) and distinct route of exposure to HIV-1 (sexual and injection drug use). Matched HIV-1 positive subjects and healthy controls were also analyzed. In all case-control cohorts the minor G allele at rs4704846 was more common in HIV-1 infected individuals than in HESN, with healthy controls showing intermediate frequency. Results from the three association analyses were combined through a random effect meta-analysis, which revealed no heterogeneity among samples (Cochrane's Q, p value = 0.89, I2 = 0) and yielded a p value of 6.8 710 124. The minor G allele at rs4704846 was found to increase HAVCR2 expression after in vitro HIV-1 infection. Thus, a positively selected polymorphism in the 3\u2032 UTR, which modulates HAVCR2 expression, is associated with the susceptibility to HIV-1 infection. These data warrant further investigation into the role of TIM-3 in the prevention and treatment of HIV-1/AIDS

Low appendicular muscle mass is correlated with femoral neck bone mineral density loss in postmenopausal women

<p>Abstract</p> <p>Background</p> <p>After menopause, rapid bone mass loss occurs in response to hypoestrogenism. Several studies suggest that muscle mass and bone mineral density (BMD) are positively associated in postmenopausal women. Therefore, it may be assumed that postmenopausal low appendicular muscle mass (aMM) can increase BMD loss in a short period of time.</p> <p>Objective</p> <p>The purpose of this study was to assess relationship of aMM with femoral neck BMD in postmenopausal women.</p> <p>Methods</p> <p>Prospective, controlled clinical Trial including 64 women aged 45-70 years, who had not had their last menstruation for at least one year. Subjects were divided into two groups: low aMM (n = 32), and normal aMM (n-32). Femoral neck BMD and muscle mass were measured by DXA at baseline and after twelve months. Pairwise and independent t tests were used for data analysis.</p> <p>Results</p> <p>Baseline weight, BMI and muscle mass (total and appendicular) significantly differ between groups (p < 0.05). After twelve months, femoral neck BMD was significantly lower in the group with low aMM, whereas no significant difference was observed in the group with normal aMM (p < 0.05).</p> <p>Conclusion</p> <p>In postmenopausal women, low appendicular muscle mass is associated negatively with femoral neck BMD in a short period of time.</p