In the peripheral blood mononuclear cells (PBMCs) of HCV infected patients the expression of STAT1 and IRF-1 is downregulated while that of caspase-3 upregulated.

Several peripheral blood mononuclear cell (PBMC) defects have been linked with hepatitis C virus (HCV) infection, including alterations in cytokine secretion and increased cell death. This study was performed to investigate the expression levels of signal transducer and activator of transcription 1 (STAT1), interferon regulatory factor 1 (IRF-1), and caspase 3 in PBMCs of patients infected with HCV. STAT1, IRF-1, and caspase 3 expression levels were compared in PBMCs from 19 untreated (naïve) HCV+ patients, 8 treated (sustained responder [SR]) HCV patients, and 20 HCV- healthy controls. Moreover, PBMCs from naïve HCV+ patients and SR-HCV patients were also evaluated for HCV RNA expression. The expression levels of STAT-1 and IRF-1 were significantly downregulated in PBMCs from naïve HCV+ patients (P  Keywords: PBMC; hepatitis C virus; STAT1; IRF-1; caspase-3.

Strategies to manage hepatitis C virus infection disease burden-Volume 4.

The hepatitis C virus (HCV) epidemic was forecasted through 2030 for 17 countries in Africa, Asia, Europe, Latin America and the Middle East, and interventions for achieving the Global Health Sector Strategy on viral hepatitis targets-"WHO Targets" (65% reduction in HCV-related deaths, 90% reduction in new infections and 90% of infections diagnosed by 2030) were considered. Scaling up treatment and diagnosis rates over time would be required to achieve these targets in all but one country, even with the introduction of high SVR therapies. The scenarios developed to achieve the WHO Targets in all countries studied assumed the implementation of national policies to prevent new infections and to diagnose current infections through screening.

Association of single nucleotide polymorphisms in microRNAs with susceptibility to hepatitis B virus infection and HBV-related liver complications: A study in a Saudi Arabian population.

The aim of this study was to evaluate the association of 10 SNPs in different microRNAs (miRNAs) with susceptibility to hepatitis B virus (HBV) infection, HBV clearance, persistence of chronic HBV infection, and progression to liver cirrhosis and hepatocellular carcinoma (HCC). Patients were categorized into the following groups: inactive HBV carrier, active HBV carrier, HBV-cleared subject and cirrhosis+HCC. Samples were analysed for 10 SNPs in microRNAs using either PCR-based genotyping or the TaqMan assay. We found that rs1358379 was associated with susceptibility to HBV infection, HBV clearance, persistent chronic HBV infection and liver cirrhosis+HCC. In addition, we found that rs2292832 and rs11614913 were associated with risk of HBV infection, viral clearance and cirrhosis+HCC, whereas rs2910164 was associated with proneness to HBV infection, and ability to clear the virus. There was evidence of associations between rs6505162 and HBV clearance and the development of liver disease, whereas a single association was found between rs2289030 and HBV clearance. Similarly, rs7372209 and rs4919510 were specifically associated with the development of HBV-induced liver complications. SNPs in miRNAs affect the susceptibility, clearance and progression of HBV infection in Saudi Arabian patients. We found, using Gene Ontology or pathway analyses, that these genes may contribute to the pathophysiology of HBV infection and related liver complications. However, differences in the association of examined SNPs with various clinical stages indicate variations in the respective functional roles of these polymorphisms and their miRNAs, and thus, further investigation to fully explore their therapeutic potential is warranted.

Sofosbuvir-Based Therapy for Genotype 4 HCV Recurrence Post-Liver Transplant Treatment-Experienced Patients.

Background and Aim. This is an open label prospective cohort study conducted at a tertiary care hospital. The primary endpoint is SVR12 in patients treated with sofosbuvir-based therapy in post-liver transplant patients with genotype 4 HCV recurrence. Methodology. Thirty-six treatment-experienced liver transplant patients with HCV recurrence received sofosbuvir and ribavirin ± peginterferon. Results. We report here safety and efficacy data on 36 patients who completed the follow-up period. Mean age was 56 years, and the cohort included 24 males and one patient had cirrhosis. Mean baseline HCV RNA was 6.2 log10 IU/mL. The majority of patients had ≥ stage 2 fibrosis. Twenty-eight patients were treated with pegylated interferon plus ribavirin in addition to sofosbuvir for 12 weeks and the remaining were treated with sofosbuvir plus ribavirin only for 24 weeks. By week 4, only four (11.1%) patients had detectable HCV RNA. Of the 36 patients, 2 (5.5%) relapsed and one died (2.75%). Conclusion. Our results suggest that sofosbuvir + ribavirin ± pegylated interferon can be utilized successfully to treat liver transplant patients with HCV recurrence.

Spontaneous clearance of hepatitis C genotype 4 after liver retransplantation.

BACKGROUND: Hepatitis C virus (HCV)-related cirrhosis remains the most common indication for liver transplantation worldwide. Graft reinfection with HCV is nearly universal, causing significant morbidity and mortality. Spontaneous clearance of HCV after liver transplantation and retransplantation is extremely rare. We report a case of spontaneous clearance of HCV genotype 4 that occurred shortly after 2nd liver transplantation. CASE REPORT: A 32-year-old female patient received a cadaveric liver transplant for HCV-related cirrhosis in 2007. She was not treated for HCV before transplantation. The patient developed biopsy-proven HCV recurrence with elevated transaminases and 65,553 IU/mL HCV RNA, genotype 4. She could not tolerate interferon-based treatment. The patient's condition progressively worsened and required a 2nd cadaveric liver transplantation in March 2013. Immunosuppression initially included steroids and Prograf, which was then switched to cyclosporine after the patient developed seizure. She developed acute cellular rejection which was readily treated with immunosuppression adjustment. HCV RNA became negative in April, which was confirmed in May 2013. CONCLUSIONS: Spontaneous clearance of hepatitis C rarely occurs after liver transplantation and is extremely rare after retransplantation. This finding may be explained by alterations in the host immune responses to HCV after transplantation. To our knowledge, this is the first case of spontaneous clearance of HCV genotype 4 after liver retransplantation.

Regional variation in organ donation in Saudi Arabia.

INTRODUCTION: There is marked regional variation in organ donation among the different regions of Saudi Arabia. Our aim was to study the dominating factors for these variations to improve organ donation in low-donation areas. MATERIALS AND METHODS: This study was a retrospective review of the Saudi Center for Organ Transplantation data for cadaveric organ donation from 2006 to 2012, with the number of cases reported, documented, consented, and harvested in various regions (northern, southern, eastern, western, and central). The region, number, and size of contributing intensive care units (ICUs), overall donation rate, and transplanted rate (potential donor and those harvested, respectively) were also reviewed. RESULTS: Between 2006 and 2012, a total of 512 cases were procured and analyzed from Saudi Arabia. From the central region, 393 were acquired, representing 76.7% of the total consented cases. These 393 cases came from 30 of 97 contributing ICUs (31%). The eastern region was ranked second, followed by the western region. The conversion rate for all regions followed a similar trend. CONCLUSIONS: There is marked variation with regard to organ donation in different regions throughout Saudi Arabia, from 1.9% in the southern region to 76.7% in the central region. This finding is related to the presence of a Mobile Action Donor Team in the central region. The number of potential donors and the contributing ICUs were strong predictors of the number of actual donors. We suggest that having a mobile donor team in each region will increase the number of donors by at least 3 times within the next 3 to 5 years.

Discriminant value of serum HBV DNA levels as predictors of liver fibrosis in chronic hepatitis B.

Current guidelines recommend antiviral therapy in chronic hepatitis B (HBV) patients with significant histological disease. We aimed to compare histological fibrosis (METAVIR, ≥F2) in patients with HBV DNA ≥20,000 IU/mL vs. ≥2000 IU/mL and identify predictors of fibrosis. We performed prospective liver biopsies on 203 HBeAg-negative patients in four groups: Group I (n = 55): HBV DNA ≥20,000 IU/mL and persistently elevated alanine aminotransferase (ALT) levels (PEALT; >40 U/L); Group II (n = 34): HBV DNA ≥20,000 IU/mL and persistently normal ALT (PNALT); Group III (n = 40): HBV DNA <20,000 IU/mL and PEALT; and Group IV (n = 74): HBV DNA <20,000 IU/mL, and PNALT. We reanalysed all groups in relation to updated cut-off for treatable viremia (2000 IU/mL). Genotype D was detected in 86% of patients. Hepatic fibrosis ≥F2 was detected in 72.7%, 52.9%, 57.5% and 18.9% in Groups I-IV, respectively (P < 0.0001). Except in Group II with a trend for lower ≥F2 fibrosis (P = 0.067), there was no significant difference by using HBV DNA cut-off 20,000 vs. 2000 IU/mL. Multivariate logistic regression analysis identified study Group IV (OR, 0.0276; CI: 0.088-0.868; P = 0.0276) and milder (A0-1) necroinflammatory grade (OR, 0.135; CI: 0.063-0.287; P < 0.0001) as independent predictors of ≥F2 fibrosis. The specificity, positive and negative predictive values for PEALT in detection of ≥F2 fibrosis for viremia ≥2000 IU/mL (80%, 69% and 65%, respectively) or ≥20,000 IU/mL (86%, 73% and 63%, respectively) were similar, with a marginal gain in sensitivity (51% vs. 42%, respectively). Significant fibrosis is prevalent in a large proportion of HBeAg-negative patients with high viremia and persistently normal ALT. Lower HBV DNA cut-offs could be adopted with marginal gains in fibrosis detection and without loss of diagnostic accuracy.

Clinical outcomes for Saudi and Egyptian patients receiving deceased donor liver transplantation in China.

Long waiting list times in liver transplant programs in Saudi Arabia and unavailability of deceased donor transplantation in Egypt have led several patients to seek transplantation in China. All patients who received transplants in China and followed in three centers from January 2003-January 2007 were included. All patients' charts were reviewed. Mortality and morbidity were compared to those transplanted in King Faisal Specialist Hospital & Research Centre (KFSH&RC) during the same period. Seventy-four adult patients were included (46 Saudi nationals; 28 Egyptians). One-year and 3-year cumulative patient survival rates were 83% and 62%, respectively compared to 92% and 84% in KFSH&RC. One-year and 3-year cumulative graft survival rates were 81% and 59%, respectively compared to 90% and 84% in KFSH&RC. Compared to KFSH&RC, the incidence of complications was significantly higher especially biliary complications, sepsis, metastasis and acquired HBV infection posttransplant. Requirements of postoperative interventions and hospital admissions were also significantly greater. Our data show high mortality and morbidity rates in Saudi and Egyptian patients receiving transplants in China. This could be related to more liberal selection criteria, use of donation after cardiac death (DCD) donors or possibly more limited posttransplant care.

Is the presence of Helicobacter pylori in dental plaque of patients with chronic periodontitis a risk factor for gastric infection?

BACKGROUND: Helicobacter pylori is considered to be a pathogen responsible for gastritis and peptic ulcers, and a risk factor for gastric cancer. A periodontal pocket in the teeth of individuals with chronic periodontitis may function as a reservoir for H pylori. OBJECTIVE: The present study was undertaken to evaluate whether the presence of H pylori in the dental plaque of patients with and without periodontitis correlates with gastric involvement. METHODS: A total of 101 patients with dyspepsia were included in the present study. Subjects were divided into periodontitis and nonperiodontitis groups. For the detection of H pylori in dental plaque, samples were collected from two teeth using a periodontal curette. Subgingival plaque was obtained by inserting two sterile paper points into periodontal pockets for 20 s. This was followed by an upper gastrointestinal endoscopy and antral biopsies. RESULTS: Sixty-five per cent of patients had dental plaque positive for H pylori and more than 50% harboured the bacteria in their stomach. Periodontitis patients had a significantly higher percentage of H pylori in their dental plaque (79% versus 43%; P<0.05) and the stomach (60% versus 33%; P<0.05) than patients with no periodontitis. Additionally, 78% of patients from the periodontitis group versus only 30% from the nonperiodontitis group had a positive test result for the coexistence of H pylori in both dental plaque and the stomach. CONCLUSION: Patients with poor oral hygiene have a higher prevalence of H pylori in dental plaque and in the stomach. This finding suggests that the oral cavity may be a reservoir for H pylori, and potentially a source of transmission or reinfection.

Acute myelomonocytic leukemia with histologic features resembling sarcomatoid carcinoma in bone marrow.

We report a case of primary acute myelomonocytic leukemia involving the bone marrow that resembled sarcomatoid carcinoma. The neoplastic cells in bone marrow biopsy specimens formed cohesive-appearing clusters and cords separated by an immature fibroblastic proliferation and myxoid stroma. Blasts in the bone marrow aspirate smears formed clusters and sheets, and a subset of blasts exhibited erythrophagocytosis. Dysgranulopoiesis was also present. Lineage was confirmed by immunohistochemical analysis of formalin-fixed, paraffin-embedded tissue. The tumor cells showed strong reactivity for lysozyme, myeloperoxidase, CD45, and CD68 and were negative for keratin, S100, CD20, and CD3. The serum lysozyme concentration (110 microgram/mL) was 13 times greater than the normal value (8 microgram/mL). Cytogenetic studies performed on bone marrow aspirate material revealed a complex karyotype, including trisomy 8 and abnormalities of chromosome 11q. We report this case of acute myelomonocytic leukemia because the neoplastic cells appeared cohesive and spindled, resembling sarcomatoid carcinoma, and therefore caused diagnostic difficulty. Other monocytic neoplasms with similar resemblance to carcinoma or sarcoma have been reported in the literature, suggesting that the tendency to appear cohesive may be an inherent characteristic of neoplastic cells with monocytic differentiation.

Gallbladder hyporesponsiveness to an exogenous nitric oxide donor, glyceryl trinitrate, in patients with advanced liver cirrhosis.

OBJECTIVE: An increased production of nitric oxide (NO) in liver cirrhosis has been documented. NO could intervene in regulating gallbladder contraction, as suggested by clinical and experimental studies. Our aim was to investigate the influence of an NO donor on gallbladder motility in cirrhotic patients in relation to the severity of liver cirrhosis. METHODS: The subjects were six controls and 18 patients with liver cirrhosis (six in each Child class). Gallbladder emptying was monitored by ultrasound for 90 min after a mixed meal (14 g fat, 425 kcal). Fasting gallbladder volume, minimal residual volume, ejection fraction, area under emptying curve, and half contraction time of the gallbladder were assessed at 15-min intervals. The patients were evaluated on two consecutive days, with or without perlingual administration of 0.5 mg glyceryl trinitrate (GTN). Statistical analysis was performed by the two-tailed Student's t test and Pearson's correlation coefficient. RESULTS: GTN significantly reduced gallbladder motility in controls and compensated cirrhotics (p < 0.02), but had no effect upon gallbladder emptying in Child class B and C cirrhotics. CONCLUSIONS: Gallbladder hypocontractility in liver cirrhosis is related to the severity of the disease. This study is the first to show that GTN has no effect upon gallbladder motility in advanced liver cirrhosis when administered in doses that induce relaxation in controls and compensated cirrhosis.

Acute and chronic actions of ethanol on endogenous calmodulin content in synaptic plasma membranes from rat brain.

The present study was designed to demonstrate that endogenous calmodulin (CaM) content in synaptic plasma membranes (SPM) is altered by acute and chronic administration of ethanol and is a sequel to the kinetic characterization of ethanol inhibition of [125I]CaM binding to SPM reported in our previous study. In rats, an acute ethanol injection (2 g/kg, i.p.) rapidly reduced CaM content in SPM from cerebral cortex, whereas chronic ethanol treatment [6% (w/v) in a liquid diet for 3 weeks] led to an up-regulation of the CaM content. In both cases, the alteration of CaM content in SPM occurred in the EGTA-dissociable pool of CaM (77% of total membrane CaM); the EGTA-nondissociable pool (23% of total CaM) was not affected. In animals receiving chronic ethanol treatment, CaM content in SPM was not altered significantly by the acute ethanol dose that produced rapid reduction of CaM content in control animals, indicating that resistance to ethanol develops. This resistance to ethanol can be attributed to alterations of membrane properties. In control SPM, ethanol at 50 mM markedly accelerated the temperature-dependent dissociation of endogenous CaM, whereas in SPM from animals chronically treated with ethanol, significant acceleration of CaM dissociation required ethanol concentrations as high as 150-200 mM. These findings on SPM in vitro were consistent with the data on CaM content obtained in vivo. Since CaM mediates a variety of biochemical processes in synaptic membranes, we hypothesize that the effects of ethanol in altering the content of membrane-bound CaM may lead to a cascade of consequences in synaptic membrane function.

Anti-C-reactive protein inhibits the calcium-dependent stage of natural killer cell activation.

The studies described in this publication were designed to determine which stage of the NK lytic mechanism is inhibited by anti-C-reactive protein (CRP). Although anti-CRP prevents target cell lysis, it does not block E:T cell conjugate formation. In parallel experiments, the number of conjugates observed in the presence of anti-CRP was normal, whereas the number of target cells killed by this same group of effector cells was greatly inhibited. Since an early stage of NK-mediated lysis requires calcium, conjugates can be synchronized by incubating effector and target cells in the absence of calcium. When conjugates were formed in the absence of calcium, and anti-CRP and calcium were then added to cultures at the same time, anti-CRP inhibited maximally. Anti-CRP continued to inhibit somewhat throughout the calcium-dependent stage but did not block lysis when added after the completion of calcium requiring events. Events that are blocked by anti-CRP must be required for the generation of NK cytotoxic factor because anti-CRP blocks the production of this factor. Once generated, however, anti-CRP does not block the activity of NK cytotoxic factor. This evidence indicates that anti-CRP blocks NK-mediated lysis at the calcium dependent stage of lysis. Events that follow this stage in the lytic process are also inhibited.

Effect of herpesvirus infections on T-lymphocyte subpopulations and blastogenic responses in renal transplant recipients receiving cyclosporine.

The immunosuppressive effects of three herpesviruses--cytomegalovirus (CMV), Epstein-Barr virus (EBV), and herpes simplex virus (HSV)--were assessed in 29 renal transplant recipients treated with cyclosporine and prednisone. The ratios of Leu 3-positive ("helper-inducer") to Leu 2-positive ("suppressor-cytotoxic") T lymphocytes in peripheral blood were only moderately and transiently decreased during primary CMV infection, with or without concurrent reactivated EBV and HSV infections. This effect was due to an increase in absolute numbers of Leu 2-phenotypic and decrease in Leu 3-phenotypic T cells and was associated with symptomatic viral illness. Reactivated CMV infection alone or together with reactivated EBV and HSV infections resulted in less significant alterations in T-cell subsets than did primary CMV infection. Lymphocyte blastogenesis was not significantly altered during the herpesvirus infections. The data suggest that cyclosporine treatment inhibits the activation of suppressor cells and depression of cellular immune function that have been associated with herpesvirus infections in renal transplant recipients undergoing conventional immunotherapy.

Lymphocyte subsets and natural killer cell responses during cytomegalovirus mononucleosis.

Cell-mediated immunity in seven patients with cytomegalovirus mononucleosis was assessed by monoclonal antibody to lymphocyte subsets, lymphocyte blastogenesis, and natural killer cell activity. The patients had decreased ratios of helper (Leu 3a) to suppressor-cytotoxic (Leu 2a) T cells as compared with normal donors. Leu 2a+ T cells from the patients were uniquely sensitive to overnight culturing, which resulted in a 2.6-fold increase in the helper/suppressor cell ratio and the appearance of an unusual third Leu 1+ 2a- 3a- T cell subset. This correlated directly with enhanced blastogenesis to concanavalin A after preculture. Base-line and interferon-boosted natural killer cell responses versus K562 cell targets during cytomegalovirus mononucleosis did not differ from those of normal donors. This suggests that certain cellular immune functions are depressed during cytomegalovirus mononucleosis in correlation with the emergence of a culture-sensitive subset of suppressor T cells. In contrast, cytotoxic lymphocytes appear to function normally and may aid in the resolution of the illness.