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Archives of Medical Laboratory Sciences - Volume:2 Issue: 1, Winter 2016

Archives of Medical Laboratory Sciences
Volume:2 Issue: 1, Winter 2016

  • تاریخ انتشار: 1394/12/11
  • تعداد عناوین: 7
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  • Solaleh Emamgholipour, Mohammad Ali Sahraian, Somayeh Shapourizadeh, Mohammad Ansari Page 1
    Background
    Little in known regarding the clinical relevance of SIRT1 in multiple sclerosis (MS). Here, we aimed to evaluate mRNA expression, protein level and enzyme activity of SIRT1 in peripheral blood mononuclear cells (PBMCs) isolated from relapsing –remitting MS patients (RRMS) and healthy controls.
    Materials And Methods
    Twenty patients with RR-MS and twenty two age- and sex-matched healthy subjects were enrolled in this case-control study. Following PBMCs isolation, mRNA expression was evaluated by real time-PCR. SIRT1 activity and SIRT1 protein level were measured using a fluorometric assay and an enzyme-linked immunosorbent assay (ELISA) respectively, in PBMC lysates.
    Results
    There was no statistically significant difference in the mRNA expression of SIRT1 (p=0.56) and its protein levels (p=0.15) between MS patients and healthy subjects. By contrast, SIRT1 enzyme activity were significantly (p=0.008) lower in RRMS patients compared with that in healthy subjects.
    Conclusion
    Our findings demonstrated that enzyme activity of SIRT1 is significantly lower in PBMCs of RRMS patients in comparison with healthy subjects. However, more investigations are essential to clarify the role of SIRT1 in MS pathogenesis.
    Keywords: enzyme activity, multiple sclerosis, pathogenesis
  • Farzaneh Sheikholeslami, Niloufar Sadat Kalaki, Seyed Abdolhamid Angaji, Sahar Eshghjoo, Ali Reza Janani, Samereh Gholami Moghadam, Mehrdad Araiinejad, Kazem Baesi, Minoo Mohraz, Asghar Abdoli Page 7
    Background
    Designing novel therapeutic agents has been a critical challenge for HIV disease.
    Materials And Methods
    In current study a DNA sequence which was encoded the Tat protein was synthesized and inserted in pET 28 vector. Vector was cloned in BL21-DE3 E. coli and cultured in TB media. After protein expression, recombinant Tat protein was purified by NTA affinity chromatography. The Tat purified protein efficiency and confirmed by SDS-PAGE and Western blot, respectively. We were immunized the camel against HIV-1 Tat recombinant protein to made a camelid antibody library. Total RNA was extracted from camel lymphocytes and VHH fragments synthesized and amplified using RT-PCR and Nested- PCR methods by special primers.
    Results
    The 350- 450 bp VHH gene fragment was produced by RT-PCR and Nested- PCR and extracted from agarose gel 1%. Then gel extraction was performed and pure fragments were inserted in HEN-4 vector by T4 DNA ligase.
    Conclusion
    The library can be applied for biopanning and isolation of nanobody against HIV-1 Tat Protein. Nanobody small size may be a useful drug for treatment of HIV disease because give them the potency of the recognizing the cryptic epitopes of tat and neutralized the virus.
    Keywords: HIV-1, Tat Protein, VHH, Nanobody
  • Samaneh Keshavarz, Mohammad Reza Shams Ardekani, Maliheh Safavi, Bahram Chahardouli, Fatemeh Nadali Page 12
    Background
    Urtica dioica is one of the medicinal herbs with many uses in treating various diseases. In some studies, its antiproliferative and apoptotic effects on cancer cell lines have been shown. Therefore, the evaluation of U. dioica effect was performed on KG-1 cell line for acute myelogenous leukemia (AML) for the first time in this study.
    Materials And Methods
    KG-1 cell line was treated by various extracts (aqueous, hydroalcoholic, chloroform and ethyl acetate) of U. dioica aerial parts and roots in different concentrations. Metabolic activity of extracts on cell line was assessed by MTT assay. To evaluate the percentage of apoptotic cells, the flow cytometry was performed by FITC Annexin V-PI apoptosis detection kit in KG-1 cell line treated with root chloroform (UDC-R) and ethyl acetate (UDE-R) extracts. The results have been reported as percentage of cell viability and IC50.
    Results
    Based on MTT results, the strongest IC50 in KG-1 cell line (219.361μg/ml) was related to UDC-R. The flow cytometric analysis showed that UDC-R and UDE-R in IC50 concentration induced early (53.6% and 57.4%, respectively) and late (27% and 33.2%, respectively) apoptosis in KG-1 cells after 24 hrs. The inhibition of cell proliferation by various extracts of U. dioica was dependent on concentration (p=0.000).
    Conclusion
    Flow cytometric analysis confirmed that UDC-R and UDE-R extracts affect on proliferation reduction of KG-1 cells by activating the apoptotic pathway.
    Keywords: Urtica dioica extract, acute myelogenous leukemia, KG-1, IC50, apoptosis
  • Sharif Maraghi, Manuchehr Naini-Kashani, Mitra Masroupour, Ali Sameri, Yasser Jeddi Page 16
    Background
    To report a case of orbital dirofilariasis from a patient in Abadan city Southwest Iran.
    Case report: A 54- year- old woman referred to Abadan oil company hospital, Southwest Iran with the symptoms of tearing, irritation, swelling, and itching of the right eye. In ophthalmoscopy, a live worm was observed in sub- conjunctival space. The worm was removed and sent to the laboratory. The worm was white in color with a length of 105 mm. Based on morphological characterization identified as Dirofilaria repns.
    Results
    We found that vitamin D3 suppresses the mRNA expression of TLR2 and TLR4 in patients with type II diabetes. TLR2 and TLR4 expression in the patients exposed to vitamin D3 were significantly decreased in comparison with patients who were not treated with vitamin D3.
    Conclusion
    This is the second report of ocular dirofilarisis from Southwestern tropical region of Iran. Increasing of human dirofilariasis may be attributed to environmental changes with global warming, humidity and increase of mosquito vectors and breeding, agricultural development and change in social conditions, traveling and outdoor living.
    Keywords: Dirofilariasis, conjunctivitis, Dirofilaria repns. Iran
  • Zahra Meshkat, Nayere Khadem Ghaebi, Mohammad Khajedaluee, Zahra Aghili, Sina Rostami Page 19
    Background
    Recurrent miscarriage is defined as two or more recurrent spontaneous miscarriages. Several causes have been suggested, among which, chromosomal abnormalities in couples is considered to have a role in this regard. However, its significance varies among different populations. The present study was carried out to evaluate the prevalence of chromosomal aberrations in couples with recurrent miscarriages in the city of Mashhad.
    Materials And Methods
    A retrospective study was performed on patient records at Medical Genetics Clinic of Imam Reza hospital in Mashhad (north-east of Iran) between 2003 and 2006.
    Results
    Of 151 records of recurrent miscarriages, 59 couples had undergone Karyotyping testing. Among those who had Karyotyping results, only one (1.7%) had chromosomal abnormality. The observed abnormality was associated with chromosome 9 inversion. The prevalence of consanguineous marriage among these couples was 59.0%.
    Conclusion
    In our study, unlike similar studies in other countries, the prevalence of chromosomal abnormalities was much lower. This could be interpreted either due to laboratory errors in our clinic or the real reduction in the association of chromosomal abnormalities with recurrent miscarriages in our population. Regarding our data, it seems that, at least among our population, costly Karyotyping testing is not necessary to predict further miscarriages or it could be limited to fewer cases having other associated factors.
    Keywords: Recurrent miscarriages, Karyotyping disorders, Chromosomal abnormalities, Iran
  • Mona Alaee, Heyder Farahani, Fathollah Mohaghegh Page 24
    Background
    It seems adipocytokines play an important role in breast cancer. Omentin-1 is a novel discovered plasma adipocytokine produced mainly in visceral adipose tissue. The aim of present study was to estimate. Circulating levels of omentin-1 in postmenopausal breast cancer patients as compare to healthy postmenopausal subjects.
    Materials And Methods
    This study consisted of 30 postmenopausal women with newly diagnosed breast cancer and 30 healthy postmenopausal subjects aged 45 to 70 years. Serum levels of omentin-1 were measured by enzyme-linked immunosorbent assay.
    Results
    The mean age of the case group was 54.3±8.3 years, and the control group was 50.3±3.2 years. The mean serum levels of omentin-1 in the case group were 73.1±29.7 ng/mL and in the control group were 108.8±65.4 ng/mL. The mean serum omentin-1 levels were found to be significantly lower in breast cancer patients (P=0.009). We also demonstrated the negative correlation of Omentin-1 with BMI in both groups (p=0.005), however, we did not find any association between age and serum omentin-1 levels (P=NS).
    Conclusion
    Our studied showed that Omentin-1 level was significantly reduced in the breast cancer patients as compare to healthy controls. Decreased omentin-1 levels may contribute to the development of cancer, however determination of its role and mechanisms needs more investigations. To sum up, our data suggest that omentin-1 can be used as a marker for early detection of breast cancer in postmenopausal women.
    Keywords: Breast cancer, omentin, serum
  • Mohammad Ali Esmaeili, Fatemeh Yari, Ali Amini, Mohammad Reza Rezvani Page 29
    This article reviews will focus on the concept and formation of micro particles (MPs) in circulation and their role in transfusion medicine and immune system. MPs are cell membrane derived vesicles which express markers of their parent cells and are found in circulation at low levels. Exact functions of MPs are unclear. In here, Physiological almost all types of circulating MPs including platelets MPs (PMPs), leukocytes MPs (LMPs), red blood cells MPs (RMPs) and endothelial cells MPs (EMPs) have been discussed. Furthermore, MPs present in plasma and blood products and their levels increase during storage. Thus, it can be stated that MPs are likely to cause transfusion reactions, particularly thrombotic complications and Transfusion-Related Acute Lung Injury (TRALI). Also, it is shown that the MPs may affect the immune system. However, to prove these, more and extensive studies both in vivo and in vitro need to be done.
    Keywords: Microparticles, platelets, transfusion medicine, adaptive immunity