Rania Zayed

Chronic liver disease (CLD) is a worldwide common pathology characterised by an inflammatory and fibrotic process leading to progressive evolution from chronic hepatitis to cirrhosis.Monocytes play a crucial role in the pathogenesis of inflammation and fibrosis in chronic liver diseases. Activated monocytes increase the expression of tissue factor, a key glycoprotein that participates in haemostatic and nflammatory processes. This study aims to assess the expression of tissue factor on activated peripheral blood monocytes in patients with hepatitis C virus (HCV)-induced CLD in relation to the degree of hepatic insufficiency and haemostatic imbalance. The current study included 60 patients with HCV induced
CLD, categorised after Child–Pugh into four groups: Child A, B, C and C during acute attack of haematemesis, 15 patients each, and 15 healthy subjectswere included as normal controls. Immunophenotype characterization was carried out by flow cytometric analysis for identification of monocytes tissue factor expression (CD142) on activated blood monocytes population (CD11b and CD14) in different groups studied. Data demonstrated significant increase (p<0.05) in the expression of each of CD11b, CD14 and CD142 revealing monocytes activation and increased expression of tissue factor
on peripheral blood monocytes in different groups of patients especially cases during acute attack of haematemesis, compared to healthy subjects. Increased monocytes tissue factor expression in patients with HCV may play a key role in the intensification of the inflammatory and immunological processes
in conjunction with activation of the coagulation mechanisms. The interaction of all these phenomena may trigger bleeding by perturbing the unstable haemostasis in frail patients with chronic liver disease.

Hemophilia is caused by a single-gene defect resulting in familial bleeding disorder. Small increase in
gene products could transform a severe form of hemophilia into a mild one. Stem cells from extrahepatic sources are being considered for clinical applications in liver cell therapy as they possess high in vitro culture potential and could be used in transplant procedures. We studied the differentiation
of bone marrow hemapoietic stem cells (BM-HSCs) from hemophilia patients' relatives into FVIII-producing hepatocytes aiming to expand patients' donor options for partial replacement of mutant liver cells by healthy cells in hemophilia A patients which could manage the severity of the bleeding disorder. BM-HSCs from hemophilic families were cultured in liquid culture containing hepatocyte growth factor for 6 days. Differentiation into hepatocytes was evaluated by alpha-fetoprotein (AFP) expression using mmunocytochemistry. Functional evaluation of transdifferentiation into hepatic lineage was done through albumin synthesis in culture supernatant using microalbumin assay kit, factor VIII activity by one stage clotting assay and expression of FVIII mRNA by RT-PCR. BM-HSCs-derived hepatocytes showed positive
AFP expression with a mean of 11 %. Functional tests performed showed their ability to produce albumin and perform FVIII activity. Also, FVIII mRNA expression was detected. Inducing the differentiation of BM-HSCs by in vitro manipulation may become a valuable tool to provide a cell source for hepatocyte transplant procedures for treatment of hemophilia patients.

Background: Cord blood is established as a source of stem cells for hemopoeitic reconstitution. Cord
blood transplants have been performed for more than 20 years now. However, cord blood stem cells
as a source for regenerative medicine is still under trial. The availability of cord blood and its banking
facilities make it a very useful source of hepatocytes for support of endstage liver disease. Cord blood
contains a number of stem cell subsets: CD34+, CD133+, and mesenchymal stem cells (MSCs).
Objectives: This study was conducted to compare between these subsets in hepatocyte
transdifferentiation efficiency. Hepatocyte lineage commitment was evaluated by alpha-fetoprotein
(AFP) expression and albumin synthesis.
Methods: Cord blood is assayed for viability. Magnetic separation was done for CD34+ve, CD133+ve
populations, MSCs were separated by culture on plastic flasks. Each cell fraction (CD34+ve,
CD133+ve and MSCs) was cultured in liquid culture containing hepatocyte growth factor for 7 days.
AFP expression was done using immunocytochemistry, albumin synthesis was measured in culture
supernatant using microalbumin assay kit.
Results: All three populations showed heptocyte transdifferentiation; although with varying
percentages. There was no statistically significant difference in AFP expression with MSCs showing
31% positivity, CD133+ve30% followed by CD34+ve showing 28.8%. Also, MSCs population
showed the highest albumin synthesis levels, followed by CD34+ then CD133+ cells.
Conclusion: Induction of hepatocyte-like cells is possible with all three stem cell subsets of the cord
blood. However, establishment of functional hepatic cells is higher in MSCs population.

Background. The response to antiviral therapy in HCV infected patients depends on several
predictive factors; however, the ability to achieve sustained virological response is still limited to around
60% of the patients infected with the HCV-4 genotype. Increased serum and intrahepatic interferon -γ
inducible protein 10 (IP-10) levels in patients with chronic hepatitis C have been described. The aim of
the work was to study the impact of pretreatment serum IP-10 level on the antiviral treatment outcome
in a group of Egyptian patients infected with HCV. Materials and methods. The study included 80 treatment
naive HCV patients. Serum IP-10 levels were determined by an enzyme linked immunosorbent assay
before therapy was introduced. Serum samples were examined twice by Real-Time PCR after complete
course of therapy for detection of HCV RNA; at the end of the antiviral therapy and six months later to
detect sustained virological response (SVR). Results. 57 patients (71%) achieved SVR while 23 (29%) patients were non-responders (NR). Pretreatment serum IP-10 levels were significantly lower in patients who achieved SVR than in NR (p=0.000). Conclusion. Low pretreatment serum IP-10 is a favorable predictive of response to antiviral HCV therapy in Egyptian patients.

INTRODUCTION:
Despite recent success after the introduction of combination therapy with interferon (IFN)-α and ribavirin, approximately 60% of patients with hepatitis C virus (HCV) genotype 4 fail to respond. Resistance to antiviral therapy remains a serious problem in the management of chronic hepatitis C. In most patients, HCV RNA could be detected in peripheral blood mononuclear cell (PBMC).
OBJECTIVE:
The authors aimed to investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C after IFN treatment, which may act as the source of HCV reinfection of hepatic cells.
METHODS:
Seventy patients with chronic hepatitis C were treated with IFN plus ribavirin for 48 weeks; they all achieved clearance of HCV RNA from serum. At the end of treatment, PBMC and serum were examined by real-time polymerase chain reaction for detection of HCV RNA. Six months later, HCV RNA in serum was monitored to detect sustained virologic response.
RESULTS:
: Analysis revealed the presence of detectable HCV RNA in the PBMC of 27% of patients despite clearance of serum HCV RNA. During follow-up, 80% of the patients who became serum HCV positive 6 months after the end of treatment had detectable level of HCV RNA in PBMC at the end of treatment.
CONCLUSIONS:
The absence of HCV in the serum of patients by the end of treatment does not exclude future viremia. The patient might still be a source of infection to others. It is strongly encouraged to test for HCV in PBMC to detect lack of response to treatment and persisting infection.