Tehran University Medical Journal

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Macrophage colony stimulating factor (M-CSF) has previously been shown to affect the differentiation of cells of the mono-nuclear phagocytic line. More recent studies indicate that M-CSF may have a role in pregnancy. In the present study, the expression of M-CSF in the human placenta was demonstrated. Placental mRNA was isolated and used as template for synthesis of complementary DNA (cDNA). The presence of M-CSF related sequences in the cDNA was shown by PCR and RT-PCR reactions in which M-CSF specific primers were used. In addition, it was shown that a 2.4 kb cDNA after electrophoresis and transfer to a nylon filter, hybridized with a digoxygenin labeled M-CSF specific probe.

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Background: Tuberculosis is still one of the most important causes of mortality and morbidity in many countries and is the second only to human immunodeficiency virus as a cause of death worldwide resulting from a single infectious agent. In 1993, the World Health Organization declared tuberculosis a global public health emergency. Conven-tional methods for the diagnosis of Mycobacterium tuberculosis (MTB) infections are time consuming, as MTB culture requires 3-8 weeks for growth. To determine the sensitivity of polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMC), we have evaluated Mycobacterium tuberculosis DNA in peripheral blood samples with PCR technique in adults with new cases of pulmonary and extra-pulmonary tuberculosis. Setting: Department of Infectious disease of Imam Khomeini Hospital, 2004- 2005, Tehran, Iran.

Methods: In this cross-sectional study, we evaluated MTB DNA extracted from 3ml citrated peripheral blood samples from 95 adults with new cases of pulmonary and extra-pulmonary tuberculosis. DNA extraction was performed using a commercial PCR kit with IS1081 primers. For prevention of cross contamination and reduction of false positives, all steps were performed under laminar hood.

Results: The 95 patients, 59 of whom were male, had a mean age 44.44 years (SD±20.26) 69 cases had pulmonary and 26 had extra-pulmonary tuberculosis. PCR was positive in 32 (33.7%) patients and negative in 63 (66.3%) cases. The overall sensitivity and accuracy of the PCR assay was 44.1% for pulmonary, 19.2% for extra-pulmonary and 10% for disseminated tuberculosis, respectively.

Conclusion: The low sensitivity of the IS1081 primer MTB-PCR assay on PBMC may pose problems for the rapid diagnosis of tuberculosis. However, further studies are needed to confirm this technique as an alternative test for the diagnosis of tuberculosis.

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Background: Mycoplasma contamination in cell cultures is considered as a major economic, research and production problem. In this study, mycoplasma-infected Vero cell lines were treated by various dilutions of ciprofloxacin and enrofloxacin in a timely manner. Removal of mycoplasma contamination from infected cell cultures was evaluated and demonstrated by polymerase chain reaction (PCR) method. Methods: This study was done from October 2013 to May 2014, in Human Rabies Vaccine Laboratory, Pasteur Institute Production and Research Complex, Tehran, Iran. Different dilutions of ciprofloxacin and enrofloxacin were used in sequential passages for treatment of infected Vero cell line. Based on lowest passages of the cell line, antibiotic treatment with ciprofloxacin and enrofloxacin was done. Amelioration of the infection and removal of mycoplasma contamination was confirmed in each step by PCR method. The technique for order of preference by similarity to ideal solution, TOPSIS method, was used to suggest the most efficient concentration of ciprofloxacin and enrofloxacin. Results: Proposed concentration of ciprofloxacin is 20 μg/ml, and in the second order is 200 μg/ml. For enrofloxacin the best proposed concentrations are 30, 300 and 3 μg/ml respectively. Ciprofloxacin and enrofloxacin and ability of them for removal of mycoplasma and also the time of treatment were verified by evaluation of the recurrence of infection through consecutive subcultures of the treated cell line. Conclusion: Our results showed that 20 μg/ml of ciprofloxacin was the dilution of choice for mycoplasma elimination followed by 200 μg/ml of ciprofloxacin. Concentrations of 3, 30 and 300 of enrofloxacin, respectively, are appropriate for mycoplasma removal. More detailed works would be needed to verify the authenticity of the proposed simple and affordable way of mycoplasma elimination.