Determining host factors contributing to the reactivation of JC virus in kidney transplant recipients.

BACKGROUND AND AIMS: The John Cunningham virus (JCV) is the established etiological agent of the polyomavirus-associated nephropathy among renal transplant recipients. In the present study, we aimed to determine the probable predictive factors leading to JCV replication in renal transplant patients. MATERIAL AND METHODS: Urine and plasma samples were collected from a total of 120 consecutive renal-transplanted patients without preliminary screening from Jan 2018 to Mar 2019. After DNA extraction, the simultaneous detection and quantification of JCV and BK polyomavirus (BKV) were conducted using a Real-time quantitative PCR method. Moreover, statistical analyses were performed using the statistical software packages, SPSS version 21. RESULTS: The prevalence of JCV viruria and viremia among renal transplant recipients were 26 (21.67%) and 20 (16.67%), respectively. A significant association was observed between the JCV and two risk factors, diabetes mellitus (P = 0.002) and renal stones (P = 0.015). The prevalence of JCV viremia among recipients who were grafted near time to sampling was significantly higher (P = 0.02). There was a statistically significant coexistence between BK and JC viruses among our patients (P = 0.029). The frequency of JCV viruria in males was reported almost three times more than in females (P = 0.005). The JCV shedding in urine was significantly associated with the tropical steroids like prednisolone acetate, which have been the standard regimen (P = 0.039). Multivariable analysis revealed duration of post-transplantation (OR, 0.89; P = 0.038), diabetes mellitus (OR, 1.85; P = 0.034), and renal stone (OR 1.10; P = 0.04) as independent risk factors associated with JCV viremia post-renal transplantation. CONCLUSION: It seems that the discovery of potential risk factors, including immunological and non-immunological elements, may offer a possible preventive or therapeutic approach in the JCV disease episodes. The results of this study may also help clarify the probable clinical risk factors involving in progressive multifocal leukoencephalopathy development.

Analysis of risk factors influencing the BK polyomavirus replication in patients with ESRD waiting for kidney transplantation.

BACKGROUND: Since the pre-transplant status affects the renal transplantation success and ultimately the survival rate, identifying the probable risk factors that increase the chance of BK virus replication in end-stage renal disease patients can be included in proposing proper surveillance guidelines during pre and post-transplantation. METHODS: A descriptive cross-sectional study was performed by collecting plasma samples from 192 ESRD patients undergoing hemodialysis for at least 3 months. Quantitative Real-time PCR assay was used to detect and measure the BK viral load. Demographic and clinical characteristics of the patients who had BK viremia were documented. RESULTS: 14 (7.3%) out of our 192 participants had BK virus viremia (95%CI 4.2%-11.6%). Demographic characteristics including etiology of ESRD and underlying diseases, mean duration and frequency of dialysis, co-infection with HBV and HCV did not affect the virus replication, since the difference between patients with BK virus viremia and BK virus negative individuals was not statistically significant. However, the statistical significance of the mean age of men with BKV and without BK virus viremia was found (OR: 3.42, P = 0.02 95%CI 0.86-13.61). Also, multiple regression analyses of some other parameters revealed that old age, high body mass index and male gender can be predictive factors of BK virus viremia in ESRD patients. CONCLUSION: Based on our findings, elderly male had higher chance of being exposed to BK virus viremia. Some other demographic characteristics such as a high BMI, old age and gender (male) can increase the risk of BK viremia in patients with ESRD prior to kidney transplantation.

Co-expression of Interleukin-17A molecular adjuvant and prophylactic Helicobacter pylori genetic vaccine could cause sterile immunity in Treg suppressed mice.

The increasing clinical significance of Helicobacter pylori (H. pylori) in human stomach cancer has led to global efforts to eradicate this pathogen. Recent studies have confirmed the importance of some cytokines such as Interleukin-18 (IL-18), Interleukin-8 (IL-8), Interleukin-17A (IL-17A) and Interleukin-22 (IL-22) in the pathogenesis of the so-called bacterium. This study was designed to compare the effects of Type 1T helper (Th1), Type 2T helper (Th2) cells, Regulatory T cells (Treg) and T helper 17 (Th17) modulatory effects on the efficacy of designed H. pylori vaccine by incorporating some molecular adjuvants in Treg competent and Treg suppressed groups. A bicistronic vector was used for simultaneous expression of codon-optimized Outer inflammatory protein a (OipA) gene and modified mice IL-18, IL-17A, IL-22 and Foxp3 (forkhead box P3) cytokines from four cassettes. Immunization of mice groups was performed using produced plasmids intradermally. Specific IgG1 and IgG2 and IgA antibody titers produced in mice were confirmed by enzyme-linked immunosorbent assay (ELISA) in sera and intestine obtained four weeks after the last immunization. After being stimulated with a mixture of both anti-CD28 mAb and H. pylori lysate, frequencies of single Interferon-Gamma (IFN-γ), single IL-17 and dual IFN-γ/IL-17-secreting T-cells were documented using dual-color FluoroSpot. The kinetics of Th1, Th2 and Th17 in the immunized animals was determined by relative quantification of IL-17A, IL-22, IFN-γ, IL-8, IL-2 and IL-4 specific mRNAs. Four weeks after bacterial challenge, quantitative colony count in the isolated and homogenized stomachs was utilized to assess the level of protective immunity among all groups. The results of immunologic assays showed that the highest cell-mediated immunity cytokines were produced in IL-17 receiving group in which the Treg responses were suppressed previously by the administration of the Foxp3 as an immunogen. In addition, potent clearance of Helicobacter pylori infection was seen in this group as well.

IRES-based co-expression of influenza virus conserved genes can promote synergistic antiviral effects both in vitro and in vivo.

Vaccination is the most effective method for the prevention of influenza virus infection. Currently used influenza vaccines that target the highly polymorphic viral surface antigens can provide protection when well matched with circulating virus strains. Antigenic drift or cyclically occurring pandemics may hamper the efficacy of these vaccines, which are chosen prior to each flu season. Therefore, a universal vaccine, designed to induce broadly cross-protective immunity against the highly conserved internal antigens M1 and nucleoprotein could provide durable protection against various influenza virus subtypes, and it could also reduce the impact of pandemic influenza, which occurs less frequently. Here, we describe a new influenza vaccine candidate in which two highly conserved antigens, nucleoprotein (NP) and matrix (M1), are simultaneously expressed from a bicistronic vector termed pIRESM1/NP. Mice were immunized intradermally four times with the pIRESM1/NP construct. The protection efficacy of the gene-based vaccine was assessed by IFN-γ and Granzyme B ELISpot assays, follow-up observation of weight loss, and survival rates of the mice groups against lethal challenges with influenza A virus subtypes H1N1 and H5N1. The group that received pIRESM1/NP showed full protection against disease following lethal challenge with H1N1 and H5N1. This group also generated significantly higher host immune cellular responses than the other groups. These results demonstrate that a DNA vaccine strategy based on co-expression of the M1 and NP proteins could provide an effective way to control influenza virus infection.

Predictive value of aspartate aminotransferase and alanine aminotransferase levels in vaginal fluid for the diagnosis of premature rupture of membranes.

BACKGROUND: Preterm premature rupture of membranes (PPROM) occurs in 3% of pregnancies and 30-40% of preterm labors are related to this problem. Early diagnosis of PPROM is very important due to its impact on pregnancy outcomes. OBJECTIVE: To determine the diagnostic value of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in vaginal fluid for the diagnosis of preterm premature rupture of membranes as a non-invasive and available test. MATERIALS AND METHODS: A total of 148 pregnant women between the 26(th)-36(th) gestational weeks were enrolled in the study. 74 patients were in PROM group and 74 in control group. AST and ALT levels in vaginal fluid were measured in each group. Mann Whitney U-test was used to compare AST and ALT levels in each group. RESULTS: The mean of AST level in vaginal fluid was 12.77±10.06 in PROM group vs. 6.91±10.92 in control group (p<0.001), while there were no significant difference between ALT levels in PROM group 1.51±3.17 and control group 0.89±1.15 (p=0.49). Optimal cut point of AST for the diagnosis of PROM was 4.5 IU/L in this study. The sensitivity, specificity, positive and negative predictive values were 82.4%, 63.5%, 69.32% and 78.33% respectively. CONCLUSION: According to the findings of this study, measurement of AST level in vaginal fluid can be used as a reliable test for diagnosis of PROM, but there is no good cut point for ALT level that can be practically used.

Survey of predictive value of 4-hour urine collection for diagnosis of proteinuria in preeclampsia.

BACKGROUND: Measuring the 24-hour urine protein ≥300 mg is the standard threshold value for diagnosis of preeclampsia. OBJECTIVE: This study was intended to determine if a patient's 4-hour urine protein correlate with the 24-hour value for diagnosis of preeclampsia. MATERIALS AND METHODS: This was a cross sectional study performed on 84 women with suspected preeclampsia due to positive urinary test strip with minimum protein content of 1+ and BP ≥140/90 at Al-zahra Educational Hospital in Rasht (Iran) from May 2007 to January 2008. Urine samples were collected within 24 hours in successive periods: The first 4-hour and the next 20-hours urine, in separate containers. The protein contents of 4-hour and 24-hour urine samples were calculated. Data were analyzed by intra-class correlation coefficient, and Receiver Operating Characteristic (ROC) curve. RESULTS: The ROC curve showed the cut-off point of 55.5 for 4-hour urine protein. The correlation between 4- and 24-hour urine protein excretions identified that most women (about 85.1%) with protein excretion rate of 300 mg/24h or more (with preeclampsia) had the same amount of protein of 55.5 or more in their 4-hour urine excretion (p<0.001). Also, most of them (about 83.7%) with a total urinary protein excretion of less than 300 mg/24h (no preeclampsia) had a protein excretion rate of less than 55.5 mg/4h. CONCLUSION: This study showed 4-hour protein collection can be used as acceptable substitute for assessing the protein content of 24-hour urine samples as a more convenient, faster, and cheaper method for diagnosis of preeclampsia and the cut-off point for 4-hour urine protein is 55.5 mg. This article extracted from a submitted thesis. (Mina Moslehi).

Perinatal outcomes of pregnancies with borderline versus normal amniotic fluid index.

BACKGROUND: Amniotic fluid is an indicator of placental function on the fetal development. The amniotic fluid index is the most commonly used method of measuring amniotic fluid. OBJECTIVE: The purpose of this study was to compare the pregnancy outcomes of a borderline versus normal AFI. MATERIALS AND METHODS: This cross-sectional study was carried out on a total of 235 pregnant women referred to Alzahra Medical Center between 2009-2011. Women with a singleton pregnancy in third trimester were enrolled into this study; of these subjects, 141 cases were in normal AFI group and 94 cases in borderline AFI group. Adequate information was obtained from the patients' medical record and the groups were compared on maternal and fetal complications. Data analysis was performed by using SPSS. RESULTS: The mean maternal age in borderline AFI group was 25.96±5.92 years and in normal AFI group was 27.88±6.5 years (p=0.023). Maternal outcomes such as preterm delivery and labor induction in women with borderline AFI were considerably higher than those in normal group (p=0.01 and p=0.001). There were no significant differences between the two groups in terms of high blood pressure, preeclampsia, diabetes and neonatal respiratory distress. The borderline AFI group had higher rate of neonatal complications such as Apgar score of less than 7 (p=0.004), IUGR (0.0001), LBW (0.001), and crucial need to NICU (0.003). CONCLUSION: Findings indicated that there are statistical differences between adverse outcomes in borderline AFI group and normal group. This article extracted from M.D. thesis. (Samira Naimian).