12 research outputs found
Intrahepatic Expression of Interferon Alpha & Interferon Alpha Receptor m-RNA can be Used as Predictors to Interferon Response in HCV and HCC Patients
Chronic hepatitis C Virus (HCV) is the leading cause of liver cirrhosis
worldwide and in Egypt. Patients with cirrhosis secondary to chronic
HCV infection are at increased risk for developing Hepatocellular
carcinoma (HCC) in which Interferon therapy is the only effective
anti-viral therapy. The current study aimed to investigate the
expression IFN-\u3b1and IFN-\u3b1Receptor genes in liver biopsies
from patients with HCV and HCC. Correlation of their expression with
the clinical, histopathological progress of the disease and the
effectiveness of IFN therapy in HCV patients after a period of 6 months
follow-up was done. Expression of IFN-\u3b1 and IFN\u3b1-Rc m-RNA was
investigated by RT-PCR using liver biopsy specimens from 30 HCV
patients including 7 patients complicated with HCC. Liver biopsies were
also subjected to formalin fixation for complete histopathological
examination. Ninety seven percent of patients expressed Interferon
Alpha m-RNA while 30% only expressed Interferon Alpha Receptor m-RNA.
Responders and non-responders to Interferon therapy were divided
according to their HCV RNA after six-months follow up period of
interferon therapy. Responders showed significantly lower mean age,
better histopathological states and higher incidence of expression of
IFN Alpha Receptor mRNA. Regardless of the response to interferon,
histological activity index scores and the degree of fibrosis showed a
significant inverse correlation to the presence of IFN\u3b1-R m-RNA.
IFN\u3b1-R mRNA expression decreases with the histological progress of
the disease, suggesting that lower expression of the IFN\u3b1-Rc may
be partially responsible for the unfavorable response to interferon in
these patients
Molecular Characterization of Chromosome 7 in AML and MDS Patients
Myelodysplastic syndromes (MDS) share many features with acute myeloid
leukemias (AML) and in fact 20 - 40% of the patients eventually develop
a picture of full blown AML. Chromosome 7 has been a focus of attention
as a site harboring tumor suppressor genes whose loss of function
contributes to leukemia transformation or tumor progression.
Abnormalities of chromosome 7 are frequently encountered in AML and
MDS. The aim of the present study was to detect the molecular
abnormalities of chromosome 7 in Egyptian AML and MDS patients using
the FISH technique and whether the abnormality has an implication on
the prognosis of the disease after a period of one year follow up.
Fluorescence in-situ hybridization (FISH) was performed for chromosome
7 using a locus specific probe for 7q31 and a centromeric probe from
7p11.1-q11.1 in a series of 30 patients diagnosed as: AML (20 patients)
and MDS (10 patients) according to the FAB criteria. Aberrations of
Chromosome 7 were found in 36.6% of AML patients: 3 cases showing
monosomy with a mean positivity of 17.3%, 2 cases showing 7q deletion
with a mean positivity of 11%. While both monosomy and deletion were
detected in 3 cases. However, in MDS patients; monosomy for chromosome
7 was the only abnormality detected and was found in 30% of cases.
Genetic abnormality of chromosome 7 showed a significant association
with poor prognostic criteria. Patients who had normal FISH results
showed a higher percentage (31.6%) of complete remission (CR) versus 0%
in patients with monosomy or deletion who showed a higher percentage
(100%) of death or poor response to therapy (NR). Although AML patients
had a worse prognosis when compared to MDS patients, patients with
genetic abnormalities showed the worst outcome
Rapid Screening of \u3b2-Globin Gene Mutations by Real-Time PCR in Egyptian Thalassemic Children
Thalassemia is one of the most common genetic disorders in Egypt. With
the total population of 70 million, there are approximately 600,000
affected individuals and more than 20 million thalassemia carriers.
Thalassemia is therefore one of the major health problems in Egypt.
B-Thalassemias are priority genetic diseases for prevention programs.
Rapid genotype characterization is fundamental in the diagnostic
laboratory, especially when offering prenatal diagnosis for carrier
couples. Introduction of the real time PCR has made a revolution in the
time taken for the PCR reactions. We present a method for the diagnosis
of the common mutations of the B-thalassemia in Egyptian children &
families. The procedure depends on the real-time PCR using specific
fluorescently labeled hybridization probes. The melting temperature for
each of the specific probes obtained after the PCR reaction permits the
identification of the specific mutation. Genotyping of 20 thalassemic
children attending the hematology clinic of the children specialized
hospital and 10 controls was done using Real-time PCR and the
conventional Amplification Refractory Mutation System (ARMS) technique.
Analysis revealed identical results to most of the patients and they
were further checked by the sequencing results of the DNA samples. The
established method is a robust, fast and straight forward assay that
allows the detection of the common B-thalassemia mutations in Egypt.
The described LightCycler system protocol can rapidly screen for many
B-globin gene mutations
Intrahepatic Expression of Interferon Alpha & Interferon Alpha Receptor m-RNA can be Used as Predictors to Interferon Response in HCV and HCC Patients
Chronic hepatitis C Virus (HCV) is the leading cause of liver cirrhosis
worldwide and in Egypt. Patients with cirrhosis secondary to chronic
HCV infection are at increased risk for developing Hepatocellular
carcinoma (HCC) in which Interferon therapy is the only effective
anti-viral therapy. The current study aimed to investigate the
expression IFN-αand IFN-αReceptor genes in liver biopsies
from patients with HCV and HCC. Correlation of their expression with
the clinical, histopathological progress of the disease and the
effectiveness of IFN therapy in HCV patients after a period of 6 months
follow-up was done. Expression of IFN-α and IFNα-Rc m-RNA was
investigated by RT-PCR using liver biopsy specimens from 30 HCV
patients including 7 patients complicated with HCC. Liver biopsies were
also subjected to formalin fixation for complete histopathological
examination. Ninety seven percent of patients expressed Interferon
Alpha m-RNA while 30% only expressed Interferon Alpha Receptor m-RNA.
Responders and non-responders to Interferon therapy were divided
according to their HCV RNA after six-months follow up period of
interferon therapy. Responders showed significantly lower mean age,
better histopathological states and higher incidence of expression of
IFN Alpha Receptor mRNA. Regardless of the response to interferon,
histological activity index scores and the degree of fibrosis showed a
significant inverse correlation to the presence of IFNα-R m-RNA.
IFNα-R mRNA expression decreases with the histological progress of
the disease, suggesting that lower expression of the IFNα-Rc may
be partially responsible for the unfavorable response to interferon in
these patients
S/MAR Element Facilitates Episomal Long-Term Persistence of Adeno-Associated Virus Vector Genomes in Proliferating Cells
Adeno- associated virus (AAV) vectors are one of the most frequently applied gene transfer systems in research and human clinical trials. Since AAV vectors do not possess an integrase activity, application is restricted to terminally differentiated tissues if transgene expression is required long term. To overcome this limitation and to generate AAV vectors that persist episomally in dividing cells, AAV vector genomes were equipped with a scaffold/matrix attachment region (S/MAR). After a mild antibiotic selection, cells transduced with AAV- S/MAR established colonies that maintained long- term transgene expression (> 50 population doublings) from replicating AAV vector episomes in the absence of further selection. Unexpectedly, with a lesser but still significant efficiency, the control vector (AAV- DS/MAR), a standard single- stranded AAV vector, also established stable transgene- expressing colonies, most of which were maintained as replicating episomes rather than integrated vector genomes. Thus, based on the result in HeLa cells, it is concluded that AAV vector genomes per se possess the ability to establish episomal maintenance in proliferating cells, a feature that can be enhanced by incorporation of a foreign genomic element such as an S/MAR element
Substantial Overview on Mesenchymal Stem Cell Biological and Physical Properties as an Opportunity in Translational Medicine
Mesenchymal stem cells (MSC) have piqued worldwide interest for their extensive potential to treat a large array of clinical indications, their unique and controversial immunogenic and immune modulatory properties allowing ample discussions and debates for their possible applications. Emerging data demonstrating that the interaction of biomaterials and physical cues with MSC can guide their differentiation into specific cell lineages also provide new interesting insights for further MSC manipulation in different clinical applications. Moreover, recent discoveries of some regulatory molecules and signaling pathways in MSC niche that may regulate cell fate to distinct lineage herald breakthroughs in regenerative medicine. Although the advancement and success in the MSC field had led to an enormous increase in the amount of ongoing clinical trials, we still lack defined clinical therapeutic protocols. This review will explore the exciting opportunities offered by human and animal MSC, describing relevant biological properties of these cells in the light of the novel emerging evidence mentioned above while addressing the limitations and challenges MSC are still facing
Direct contact of umbilical cord blood endothelial progenitors with living cardiac tissue is a requirement for vascular tube-like structures formation
The umbilical cord blood derived endothelial progenitor cells (EPCs) contribute to vascular regeneration in experimental models of ischaemia. However, their ability to participate in cardiovascular tissue restoration has not been elucidated yet. We employed a novel coculture system to investigate whether human EPCs have the capacity to integrate into living and ischaemic cardiac tissue, and participate to neovascularization. EPCs were cocultured with either living or ischaemic murine embryonic ventricular slices, in the presence or absence of a pro-angiogenic growth factor cocktail consisting of VEGF, IGF-1, EGF and bFGF. Tracking of EPCs within the cocultures was performed by cell transfection with green fluorescent protein or by immunostaining performed with anti-human vWF, CD31, nuclei and mitochondria antibodies. EPCs generated vascular tube-like structures in direct contact with the living ventricular slices. Furthermore, the pro-angiogenic growth factor cocktail reduced significantly tubes formation. Coculture of EPCs with the living ventricular slices in a transwell system did not lead to vascular tube-like structures formation, demonstrating that the direct contact is necessary and that the soluble factors secreted by the living slices were not sufficient for their induction. No vascular tubes were formed when EPCs were cocultured with ischaemic ventricular slices, even in the presence of the pro-angiogenic cocktail. In conclusion, EPCs form vascular tube-like structures in contact with living cardiac tissue and the direct cell-to-cell interaction is a prerequisite for their induction. Understanding the cardiac niche and micro-environmental interactions that regulate EPCs integration and neovascularization are essential for applying these cells to cardiovascular regeneration