Viruses can alter the expression of host microRNAs (MiRNA s) and modulate the immune response during a persistent infection. The dysregulation of host MiRNA s by hepatitis B virus (HBV) contributes to the proinflammatory and profibrotic changes within the liver. Multiple studies have documented the differential regulation of intracellular and circulating MiRNA s during different stages of HBV infection. Circulating MiRNA s found in plasma and/or extracellular vesicles can integrate data on viral-host interactions and on the associated liver injury. Hence, the detection of circulating MiRNA s in chronic HBV hepatitis could offer a promising alternative to liver biopsy, as their expression is associated with HBV replication, the progression of liver fibrosis, and the outcome of antiviral treatment. The current review explores the available data on miRNA involvement in HBV pathogenesis with an emphasis on their potential use as biomarkers for liver fibrosis.

HIV-HBV co-infection was previously studied in patients older than 40 years of age, with added risk factors for liver disease. Data on progression of liver damage in young, co-infected patients, without associated comorbidities is sparse. We assessed the prevalence and stage of HBV co-infection in young individuals with parenterally acquired HIV infection and multiple previous antiretroviral (ART) treatments. Out of 227 patients (median age: 24 years, median duration of HIV infection: 23.9 years, median ART duration: 11.6 years), 61.7% had any markers of HBV infection, 53.6% were chronic HBsAg carriers, 9.3% had HBeAg present, 11.4% had high-levels of HBV DNA and 1.4% had severe liver fibrosis according to FIB-4 noninvasive score. HBV co-infected patients had longer ART exposure (147.07 vs. 121.3 months, p = 0.003) than those with HIV in the absence of HBV, with no significant differences in the use or duration of HIV-HBV dually-active antiretrovirals (lamivudine, tenofovir). Young HIV-HBV co-infected patients remain free of hepatic disease under ART.

HIV-associated neurocognitive disorders (HAND) continue to be reported even in patients with successful antiretroviral treatment. We investigated the prevalence of neurocognitive impairment and possible HIV-associated determinants of cognition in a Romanian cohort of young adults, parenterally infected with HIV during their first years of life. Two hundred fourteen treatment-experienced HIV-positive individuals [median age: 24 years, males: 48%, median duration on combined antiretroviral therapy (cART): 12 years] underwent standard immunologic and virological monitoring and antiretroviral resistance testing using pol gene sequencing in both plasma and, when available, cerebrospinal fluid (CSF) paired samples. Neurocognitive impairment was assessed using a comprehensive neuropsychological test battery, and a global deficit score (GDS) was calculated (cutoff ≥0.5). Cognitive impairment was detected in 35% of the study participants, without any association with sex, median age, CD4 cell count (actual or nadir), CSF and plasma viral load (actual or zenith), AIDS diagnosis, duration of HIV infection, and cART characteristics. Participants carrying resistant viruses tended to be more frequently cognitively impaired (p = 0.36), with a higher median GDS value (p = 0.06) compared with participants harboring wild-type HIV, although the figures did not reach statistical significance. No signs of virological compartmentalization were observed based on CSF versus plasma viral load and on the profile of pol sequences. A moderate rate of mild neurocognitive impairment is still present in young adults with chronic HIV infection acquired in early childhood despite successful cART, without any association with classic markers of HIV infection. New biomarkers reflecting persistent central nervous system inflammation and neuronal injury may be more relevant for the development of HAND.

Small RNAs and their associated RNA interference (RNAi) pathways underpin diverse mechanisms of gene regulation and genome defense across all three kingdoms of life and are integral to virus-host interactions. In plants, fungi and many animals, an ancestral RNAi pathway exists as a host defense mechanism whereby viral double-stranded RNA is processed to small RNAs that enable recognition and degradation of the virus. While this antiviral RNAi pathway is not generally thought to be present in mammals, other RNAi mechanisms can influence infection through both viral- and host-derived small RNAs. Furthermore, a burgeoning body of data suggests that small RNAs in mammals can function in a non-cell autonomous manner to play various roles in cell-to-cell communication and disease through their transport in extracellular vesicles. While vesicular small RNAs have not been proposed as an antiviral defense pathway per se, there is increasing evidence that the export of host- or viral-derived RNAs from infected cells can influence various aspects of the infection process. This review discusses the current knowledge of extracellular RNA functions in viral infection and the technical challenges surrounding this field of research. This article is categorized under: Regulatory RNAs/RNAi/Riboswitches > Regulatory RNAs RNA in Disease and Development > RNA in Disease Regulatory RNAs/RNAi/Riboswitches > RNAi: Mechanisms of Action.

Objectives: The study aimed to investigate the role of miR-125b as a non-invasive biomarker in chronic hepatitis C. Methods: An observational study was conducted on 94 treatment-naive HCV-infected patients (mean age 49.8 +/- 11.5 years, 59.6% females). Liver fibrosis was assessed by transient elastography (TE) and the expression of miR-125b in plasma was quantified by real-time PCR. Results: All patients were infected with HCV genotype 1b and had active viral replication, 42.6% had significant cytolysis, and 73.4% had increased serum gamma-glutamyl transferase (GGT) values. Significant fibrosis (liver stiffness measured by TE of > 7.1 kPa) was present in 61.7% of the patients. No significant associations were found between miR-125b expression and baseline HCV viral load (P = 0.56), IL28B polymorphisms (P = 0.5), alpha-fetoprotein levels (P = 0.27), and patients' gender (P = 0.13) or age (P = 0.5). In a univariate analysis, the miR-125b expression level was significantly correlated with ALT (P = 0.001) and GGT levels (P < 0.0001). An up-regulated expression of miR-125b was found in plasma samples from patients with advanced liver fibrosis as compared to those with mild/moderate fibrosis [mean miR-125b value = 0.002 versus 0.001(P = 0.02)]. In a multiple regression analysis, an upregulated miR-125b expression level remained independently in association only with significant fibrosis and increased GGT level (P= 0.026; R-2 = 0.242). Conclusions: An up-regulated miR-125b expression might be an indicator of severe liver fibrosis in patients with chronic hepatitis C, independent of the viral replication level.

Matrix metalloproteinases (MMPs) are known ezymes involved in the modulation of extracellular matrix (ECM) and adipocyte and preadipocytes differentiation. Obesity implies a more or less rapid but generalized increase in adipose tissue (adipocyte and preadipocytes) and this proceses generate abnormal ECM metabolism. Aim: This study purpose was to determin if we can consider MMPs as a marker of effectiveness in bariatric surgery. Materials and methods: The experimental study used 20 obese wistar rats (10 in control group and 10 in Study group).The study group had gastric by-pass for obesity, and there were analised the pre and post operative MMP-2 and MMP-9. We compared the results in order to see if bariatric surgery modifies the MMPs status and if there is a correlation between the weight loss and the values of MMPs. Results: The MMP-2 and MMP-9 activities were detectable, but MMP-2 activity was significantly higher than MMP-9. MMP-9 was strongly correlated with body weight parameters before surgery, as well as after significant body weight reduction as a result of bariatric surgery. There is a strong correlation between a greater weight loss and the values of MMP-2. Conclusions: MMP-2 and MMP-9 are the two most important proteins of ECM involved in adipose tissue remodeling after bariatric surgery. Although we believe that more in-depth studies are required, MMPs may be considered a marker of the effectiveness of bariatric surgery and weight loss.

Abstract Aim: To investigate peripheral blood monocytes/macrophages (Mo/Mᴓ) paraoxonase 2 (PON2) in diabetes and the factors modulating its activity. Methods: One hundred and eighteen patients with newly diagnosed uncomplicated type 2 diabetes mellitus were compared regarding clinical, biochemical and oxidative stress parameters with 80 healthy subjects. The capacity of the peripheral blood mononuclear cells (PBMNC) to release pro-oxidants and to neutralise them was determined by measuring the respiratory burst (RB) and the intracellular antioxidant enzyme PON2. In vitro experiments were conducted on a differentiated monocytes cell line (dU937) that was exposed to serum deprivation followed by addition of isolated lipoproteins (VLDL or LDL). Results: Paraoxonase 2 activity in Mo/Mᴓ was significantly lower in type 2 diabetes patients (0.042 ± 0.044 vs 0.165 ± 0.133U lactonase activity/mg protein in controls, p < .0005) and decreased in the obese in all groups. It was inversely correlated to parameters of adiposity (BMI and Waist Circumference), of glucose control (blood glucose, fructosamine and HbA1c) and insulin resistance (HOMA-IR). In multivariate regression models, 15–34% of the PON2 variance was explained by diabetes. The in vitro addition of VLDL normalised the RB of serum deprived dU937 cells, S− (to 82 ± 18% of the cells incubated with serum, S+) and PON2 activity (from 0.524 ± 0.061 in S − to 0.298 ± 0.048 U/mg protein). In contrast, when LDL was added, the RB remained lower (61 ± 12% of S+, p = .03) and PON2 higher (0.580 ± 0.030 U/mg protein, p = .003). Conclusions: The decrease in monocyte/macrophage PON2 enzymatic activity observed in type 2 diabetes cannot be totally explained by abdominal obesity and insulin resistance. The underlying molecular mechanisms need to be identified.

This study aimed to investigate the influence of antiretroviral therapy on methylation markers, in a group of HIV infected, heavily treated patients. Immune and molecular methods were used to investigate potential changes in methylation profile in DNA isolated from peripheral blood mononuclear cells collected from antiretroviral-experienced HIV infected patients and healthy controls. The percentage of 5-methylcytosine was inversely correlated with proviral DNA and active replication while DNMT1 (p = 0.01) and DNMT3A (p = 0.004) independently correlated with active viral replication. DNMT3A expression increased with total treatment duration (p = 0.03), number of antiretroviral drugs ever used (p = 0.003), and cumulative exposure to protease inhibitors (p = 0.02) even in currently HIV undetectable patients.

The diabetes epidemic nowadays appeared at the same time with the obesity epidemic their relationship being a causal one. The protection offered by paraoxonase 2 (PON2), a recently discovered member of the paraoxonase multigene family that has been proposed to have antioxidant properties, is still a matter of debate. We aimed to investigate if antioxidant enzyme PON2 is affected by glucose (glucotoxicity) and lipids (lipotoxicity) levels in beta-pancreatic cells and if these, in turn, are related to functional changes in the synthesis and secretion of insulin. We determined the PON2 in beta-pancreatic cell lines INS-1 (rat) and PANC-1 (human). Cells (2X10(4) cells) were incubated with different concentrations of D-glucose (no glucose; hypo and hyperglycemia) in the presence or absence of either oleic acid (50 mu M), palmitic acid (1mM) or a mixture of them (oleate/palmitate:2/1). The cell lysates and media were separated by Tricine-SDS-PAGE and the insulin and PON2 were visualized by Western Blot and/or ELISA. With our model of in vitro islets, we showed that intracellular insulin accumulation was lower at higher glucose concentrations and this was associated with insulin secretion and PON2 changes in all cell lines studied. The effect was amplified by adding oleic acid in a combination with palmitic add. Our findings suggest that PON2 is present in beta-pancreatic cells and could protect against gluco and lipotoxicity being useful for prevention of type 2 diabetes obesity associated complications.

MicroRNAs (miRNAs) are small, non‐coding RNA species essential for the post‐translational regulation of gene expression. Several miRNA have been proposed to contribute to Human immunodeficiency virus‐1 (HIV‐1) infection establishment, progression and latency. Among them, miR‐29a seems to be of particular interest. The aim of this study was to investigate the association between miR‐29a expression and immunologic and virologic markers of HIV infection progression in long‐term antiretroviral‐treated individuals. In a homogenous group of 165 young adults, with chronic HIV infection, parenterally acquired during childhood, the expression level of miR‐29a was found to be inversely correlated with HIV viral load and the degree of immunosuppression, expressed by both CD4 cell count and the CD4/CD8 ratio. There was a significant difference in miR‐29a expression according to the patient's response to treatment, with the lowest levels expressed by patients with treatment failure, defined as detectable viremia and CD4 < 350 cells/mm3. No significant correlation was found between miRNA level and the nadir CD4 count or zenith HIV viral load. This study establishes the association between miR‐29a expression and markers of HIV infection in long‐term survivors, treatment‐experienced patients, suggesting its potential use as an indicator for the on‐treatment disease evolution. J. Med. Virol. 88:2132–2137, 2016. © 2016 Wiley Periodicals, Inc.

Objectives: Surgical treatments of obesity have been shown to produce rapid improvements in insulin sensitivity. The aim of this study was to investigate the oxidative stress status and adipokines in obese patients with type 2 diabetes mellitus (T2DM) from bariatric surgery.

MicroRNA (miRNA) are small- 19-24 nucleotides, non-coding RNA molecules that regulate translational and post-translational processes through mRNA degradation and protein translation repression, or sometimes through heterochromatin formation or activation of protein translation. Lately, miRNA are investigated as predictive biomarkers for the evolution and prognosis of viral diseases, as well as therapeutic targets. Although the role of non-coding RNA molecules during HIV infection is not yet fully elucidated, several studies have reported strong correlations between cellular and viral miRNA expression and the immunologic and virological status of infected patients. Some studies have proven the existence of host cellular miRNA able to influence all important steps in HIV replicative cycle and to interfere with the establishment of latent infection in CD4+ cells. Although the function and existence of viral encoded miRNA remains controversial, new studies have shown their potential in modulating the host cell response or the efficiency of viral replication. This review aims to summarize the current level of knowledge in the interaction between miRNA and HIV-1 and to describe new therapeutic strategies entailing miRNAs as new and potent players in controlling viral infectivity, replication and latency.

Hepatitis B virus (HBV) infection is common in individuals infected with human immunodeficiency virus, and coinfection is associated with higher rates of HBV replication and more rapid liver disease progression than HBV monoinfection. This study evaluates the prevalence and virological profiles of hepatitis B infection in a cohort of long term survivors, with multiple antiretroviral treatments. 164 HIV-infected subjects (median age: 24 years) on combined antiretroviral therapy (cART) (median duration: 13 years), were evaluated for serologic markers of HBV infection (HBsAg, total anti-HBc and anti-HBsAg antibodies). Markers of HBV infectivity (HBeAg and HBV DNA) were evaluated in all HBsAg carriers; HBV genotype and lamivudine resistance mutations were analyzed in the cases with HBV DNA >103 IU/mL. 65.9% of the patients (108/164) had markers of past or present HBV infection (antiHBc positives), out of which 51.8% (56/108) were chronic HBV carriers and 30.5% had resolved HBV infection. All subjects were equally exposed to HBV infection, irrespective of their current immune status. Out of 21 patients with isolated anti-HBc antibodies, only 4 had detectable HBV DNA, presumably having occult hepatitis B. HBV chronic carriage rate was not influenced by the immune status. Overall, only 17.8% of the chronic carriers had active HBV replication; severely immune-depressed patients tend to maintain active viral replication more frequently than those with moderate or absent immunosuppression. The majority of the coinfected individuals (68.3%) showed no sign of liver fibrosis (APRI score 1.5); HBV DNA was directly correlated with APRI score. HBV genotype A was present in all but one of the tested patients. 98.8% of the coinfected subjects have been treated with a cART regimen that includes a drug dually active against HIV and HBV (in 98% of the cases lamivudine (3TC), for a mean time of 6.9 years and in 29.7% of the cases the current dually active drug was tenofovir). 3TC-resistance mutations were present in only 4 coinfected subjects. We found a strikingly low percentage of long term HIV/HBV coinfected patients from our group with active liver disease. A high prevalence of asymptomatic HBV chronic carriage was associated with a good immune status, suggesting that dually active antiretrovirals have an important role in delaying progression of liver disease in HIV/HBV coinfected patients.

Background. The pro-thrombotic potential of the anabolic androgenic steroids (AAS), worldwide misused substances, has increasingly become a subject of current interest. Conversely, taurine, a sulfur-amino acid ubiquitous in human body, in addition to other beneficial effects, is thought to have an inhibitory effect on platelet aggregation.Purpose. To assess platelet aggregation both taurine and high doses of AAS were simultaneously chronically administered in rats.Methods. The experiment was conducted on 40 male Wistar rats, divided into 4 equal groups: control (C) - no treatment; AAS (A) treated with 10 mg/kg/week of nandrolone decanoate (DECA); taurine (T) - daily treated with oral supplementation of 2% taurine in drinking water; androgen and taurine group (AT) - concomitant administration of DECA and taurine. After 12 weeks of treatment, blood samples were collected and platelet aggregation induced by ADP was performed using the turbidimetric method.Results. The platelet aggregation magnitude was significantly higher (p<0.001) in group A (62.1 +/- 6.10%) than in group C (47.8 +/- 5.39%), while in group T (40.3 +/- 6.49%) it was significantly lower (p=0.04). Moreover, the platelet aggregation response was significantly lower in group AT (54.5 +/- 6.38%) than in group A (p=0.04), without a significant difference between group AT and group C (p=0.08).Conclusion. Our findings provide additional evidence regarding harmful potential of high doses of DECA, chronically administered. The increased platelet aggregation induced by AAS may be decreased by diet supplementation with taurine.