Nidaa Ababneh

UDP-glucuronosyltransferase 1A4 (UGT1A4) polymorphisms in a Jordanian population

Glucuronidation is one of the most important phase II metabolic pathways. It is catalyzed by a family of UDP-glucuronosyltransferase enzymes (UGTs). One of the subfamilies is UGT1A. Allele frequencies in UGT1A4 differ among ethnic groups. The aim of this study was to determine the allelic frequency of two most common defective alleles: UGT1A4*2 and UGT1A4*3 in a Jordanian population. A total of 216 healthy Jordanian Volunteers (165 males and 51 females) were included in this study. Genotyping for UGT1A4*1, UGT1A4*2 and UGT1A4*3 was done using a well established polymerase chain reaction–restriction fragment length polymorphism test. Among 216 random individuals studied for UGT1A4*2 mutation there were 26 individuals who were heterozygous, giving a prevalence of 12% and an allele frequency of 6.5%. Only one individual was homozygous for UGT1A4*2. The UGT1A4*3 mutation was detected as heterozygous in 9 of 216 individuals indicating a prevalence of 4.2% and allele frequency of 3.5%. Three individuals were homozygous for the UGT1A4*3 indicating a prevalence of 1.4%. The prevalence of UGT1A4*2 is similar to the Caucasians but different from other populations whilst the UGT1A4*3 prevalence in the Jordanian population is distinct from other populations. Our results provide useful information for the Jordanian population and for future genotyping of Arab populations in general.

Identification of atypical PML-RARA breakpoint in a patient with acute promyelocytic leukemia.

Acute promyelocytic leukemia (APL) of the M3 subtype is characterized by translocation t(15;17) that generates the PML-RARA fusion gene. Depending on the breakpoint position in the PML gene, 3 main fusion transcripts usually result. These breakpoints are bcr1 and bcr3 in introns 6 and 3, respectively, and bcr2 in exon 6. This report describes a rare atypical bcr2 breakpoint in a patient with morphological, cytogenetic and molecular features of APL. The presence of t(15;17) was first revealed by fluorescent in situ hybridization. Molecular analysis by reverse transcription polymerase chain reaction using primers for different PML-RARA junctions showed bands with different sizes compared with those generated from the three classical breakpoints, namely bcr1, bcr2 and bcr3. However, sequence analysis confirmed the presence of a bcr2 transcript with an atypical breakpoint within exon 6. The patient responded well to treatment and is now in complete remission. However, suggesting a favorable prognosis associated with such a rare transcript is difficult as more similar cases are needed to confirm such a conclusion. This article also stresses the importance of sequencing unusual polymerase chain reaction products to confirm their nature.

Allele and Genotype Frequencies of the Polymorphic Methylenetetrahydrofolate Reductase and Colorectal Cancer among Jordanian Population

BACKGROUND Methylenetetrahydrofolate reductase (MTHFR) is involved in DNA synthesis and repair. We here aimed to investigate two common polymorphisms, C677T and A1298C, with genotype and haplotype frequencies in colorectal cancer (CRC) cases among Jordanian. MATERIALS AND METHODS 131 CRC cases were studied for MTHFR C677T and A1298C polymorphisms, compared to 117 controls taken from the general population, employing the PCR-RFLP technique. RESULTS We found the frequency of the three different genotypes of MTHFR C677T among Jordanians to be CC: 61.7%, CT: 35.2%, and TT 3.1% among CRC cases and 50.9%, 38.8% and 10.3% among controls. Carriers of the TT genotype were less likely to have CRC (OR=0.25; 95%CI: 0.076-0.811; p=0.021) as compared to those with the CC genotype. Genotype analysis of MTHFR A12987C revealed AA: 38.9%, AC: 45%, and CC 16% among CRC cases and 37.4%, 50.4% and 12.2% among controls. There was no significant association between genetic polymorphism at this site and CRC. Haplotype analysis of MTHFR polymorphism at the two loci showed differential distribution of the TA haplotype (677T-1298A) between cases and controls. The TA haplotype was associated with a decreased risk for colorectal cancer (OR=0.6; 95% CI: 0.4-0.9, p=0.03). CONCLUSIONS The genetic polymorphism of MTHFR at 677 and the TA haplotype may modulate the risk for CRC development among the Jordanian population. Our findings may reflect an importance of genes involved in folate metabolism in cancer risk.

Allele and Genotype Frequencies of the Polymorphic Methylenetetrahydrofolate Reductase and Lung Cancer in ther Jordanian Population: a Case Control Study.

BACKGROUND Methylenetetrahydrofolate reductase (MTHFR) is involved in amino acid synthesis and DNA function. Two common polymorphisms are reported, C677T and A1298C, that are implicated in a number of human diseases, including cancer. OBJECTIVE The association between MTHFR C677T and A1298C genotype and haplotype frequencies in risk for lung cancer (LC) was investigated in the Jordanian population. MATERIALS AND METHODS A total of 98 LC cases were studied for MTHFR C677T and A1298C polymorphisms, compared to 89 controls taken from the general population, employing the PCR-RFLP technique. RESULTS The frequency of the genotypes of MTHFR C677T among Jordanians was: CC, 59.6%, CT, 33%; and TT, 7.4% among LC cases and 49.4%, 40.2% and 10.3% among controls. No significant association was detected between genetic polymorphism at this site and LC. At MTHFR A12987C, the genotype distribution was AA, 29.5%; AC, 45.3%, and CC 25.3% among LC cases and 36.8%, 50.6% and 12.6% among controls. Carriers of the CC genotype were more likely to have LC (OR=2.5; 95%CI: 1.04-6; p=0.039) as compared to AA carriers. Smokers and males with the CC genotype were 9.9 and 6.7 times more likely to have LC, respectively (ORsmokers=9.9; 95%CI: 1.2-84.5, p=0.018; ORmen=6.6; 95%CI: 1.7-26.2, p=0.005). Haplotype analysis of MTHFR polymorphism at the two loci showed differential distribution of the CC haplotype (677C-1298C) between cases and controls. The CC haplotype was associated with an increased risk for lung cancer (OR=1.6; 95% CI: 1.03-2.4, p=0.037). CONCLUSIONS The genetic polymorphism of MTHFR at 1298 and the CC haplotype (risk is apparently lower with the C allele at position 677) may modulate the risk for LC development among the Jordanian population. Risk associated with the 1298C allele is increased in smokers and in males. The results indicate that a critical gene involved in folate metabolism plays a modifying role in lung cancer risk, at least in the Jordanian population.

Fatal encephalitis due to BK virus in a patient with common variable immunodeficiency: a case report.

Encephalitis due to BK virus is a rare condition. Here, we describe a young male patient with common variable immunodeficiency who developed fatal encephalitis due to BK virus. The patient presented initially with ocular symptoms that were followed by behavioral changes and spastic quadriparesis. Diagnosis was made by the compatible clinical findings and detection of viral DNA by polymerase chain reaction in the cerebrospinal fluid. To the best of our knowledge, this is the first report of BK virus encephalitis in a patient with common variable immunodeficiency. We suggest that BK virus should be suspected in cases of encephalitis; particularly in patients with immunodeficiency.

Physicochemical and Microstructural Characterization of Injectable Load-Bearing Calcium Phosphate Scaffold

Injectable load-bearing calcium phosphate scaffolds are synthesized using rod-like mannitol grains as porogen. These degradable injectable strong porous scaffolds, prepared by calcium phosphate cement, could represent a valid solution to achieve adequate porosity requirements while providing adequate support in load-bearing applications. The proposed process for preparing porous injectable scaffolds is as quick and versatile as conventional technologies. Using this method, porous CDHA-based calcium phosphate scaffolds with macropores sizes ranging from 70 to 300 μm, micropores ranging from 5 to 30 μm, and 30% open macroporosity were prepared. The setting time of the prepared scaffolds was 15 minutes. Also their compressive strength and e-modulus, 4.9 MPa and 400 MPa, respectively, were comparable with those of the cancellous bone. Finally, the bioactivity of the scaffolds was confirmed by cell growth with cytoplasmic extensions in the scaffolds in culture, demonstrating that the scaffold has a potential for MSC seeding and growth architecture. This combination of an interconnected macroporous structure with pore size suitable for the promotion of cell seeding and proliferation, plus adequate mechanical features, represents a porous scaffold which is a promising candidate for bone tissue engineering.

Prevalence of blood-borne viral infections among autopsy cases in Jordan

Background: Morgues are high risk areas for the spread of infection from cadavers to staff during the post-mortem examination. Infection can spread from corpses to workers by airborne transmission, by direct contact, or through needle and sharp object injuries. Objective: Knowledge about the prevalence of these infections on autopsy is essential to determine the risk of transmission and to further enforce safety measures. Methods: This is a descriptive study. All autopsies performed in the Department of Forensic Medicine at Jordan University Hospital during the study period were tested for the serology of human immunodeficiency, hepatitis B and C viruses. Positive tests were confirmed by nucleic acid testing. Results: A total of 242 autopsies were tested. Age ranged from 3 days to 94 years (median 75.5 years, mean 45.3 years (21.9 ± SD)). There were 172 (71%) males. The cause of death was considered natural in 137 (56.6%) cases, accidental in 89 (36.8%), homicide in nine (3.7%), suicide in four (1.7%), and unknown in three (1.2%) cases. Hepatitis B surface antigen was positive in five (2.1%) cases. Hepatitis C virus antibody was positive in five (2.1%) cases and the hepatitis C virus polymerase chain reaction was positive in two (0.8%) cases. HIV antibody was not detected in any of the cases. The infection status of cases was not associated with age, sex, nationality, or cause of death. Conclusion: The study findings indicated that there is a low prevalence of virus-infected autopsies in Jordan. However, the risk of transmission remains a potential threat and therefore the necessary precautions should always be taken during autopsy.

Evaluation of the growth and osteogenic differentiation of ASCs cultured with PL and seeded on PLGA scaffolds

Abstract Scaffold serves as an important component of tissue engineering, which facilitates cell attachment, proliferation and differentiation of cultured cells. In this study we aimed to use platelet lysates as a substitute for FBS in culturing and proliferation of human adipose tissue-derived stromal cells (ASCs), which constitute a promising source for cell therapy. We characterized ASCs in the presence of PL, and then we seeded them onto poly(lactic-co-glycolic acid) (PLGA) scaffolds, osteogenic media was used to induce their proliferation and osteogenic differentiation. Gene expression analysis revealed higher expression of osteogenic related genes, immunohistochemical staining showed proper cell attachment, growth and collagen matrix formation with the ability to induce vascularization. In conclusion, expansion of ASCs in PL-supplemented medium could promote cell proliferation and osteogenic differentiation of cells seeded on PLGA scaffolds, therefore it could be considered as a suitable and effective substitute for FBS to be used in clinical applications.

ABL kinase domain mutations in patients with chronic myeloid leukemia in Jordan

Mutations of the BCR-ABL tyrosine kinase domain constitute a major cause of resistance to tyrosine kinase inhibitors in patients with chronic myelogenous leukemia (CML). In this study, we analyzed peripheral blood samples from 185 Jordanian CML patients for ABL mutations, who were on imatinib for a minimum of 6 months regardless of their disease status and over a period of 5 years. Mutations were detected by nested RT-polymerase chain reaction, followed by direct sequencing of the ABL kinase domain. Twelve different point mutations were detected 25 times in 21 patients. The resultant mutations were as follows: four patients have T315I, three of each of the following: L248V, F317L, and G250E, two of each of the following: H396R, M244V, and T277A, and one of each of the following: F311I, M318T, Q252H, F359A, F359I, and Y326H. After patient follow-up, the mutation had disappeared in 12 patients; 3 patients died; 3 patients were not retested; and 3 patients had persistent mutation. The finding of our study is in line with what has been described in the literature. Detecting ABL mutations in chronic phase may lead to positive outcome by modifying treatment.

Aptamers: Promising Molecules for Cancer Stem Cell Targeting

The term Cancer Stem Cells (CSCs) has been coined to refer to a subpopulation of tumor cells that has the ability to self-renew and to generate the diverse cell pool of a given tumor. In the last decade CSCs have been receiving a lot of interest due to their cancer initiating and maintaining capabilities making them the real driving force within a malignant mass that pushes towards more aggressive proliferation and more resistance against anticancer drugs. These cells have been linked to different specific markers in an attempt to achieve efficient isolation and characterization. Such markers include CD44, EpCAM and CD133 among others. Aptamers are synthetic single stranded oligonucleotides selected from a huge pool of random sequences which can fold and bind to a wide range of targets with high affinity and specificity. They are not immunogenic or toxic and have good clearance rates, qualities that make aptamers a good rival to monoclonal antibodies in their diagnostic as well as therapeutic applications. This review explores the potential applications of aptamers selected to target the main markers of CSCs. Such applications include diagnostic assays and analytical platforms where such aptamers can be utilized to detect and characterize the behavior of these cells in addition to predicting and monitoring disease course and response to different treatment regimens. The potential therapeutic applications are also discussed in details, where such anti- CSC aptamers can be used in a variety of formulations to efficiently target the initiating cores of tumors.