Detection of Mycoplasma species in cell culture by PCR and RFLP based method: effect of BM-cyclin to cure infections

PURPOSE: A two-stage nested polymerase chain reaction (PCR) assay system was described that amplifies the 16S-23S rRNA spacer region sequences of Mycoplasma and Acholeplasma infections in cell cultures and virus stocks. METHODS: Established cell lines and virus stocks were screened for the presence of Mycoplasma by using nested PCR using two sets of outer and inner primers, amplifies 16S-23S rRNA. PCR and restriction fragment length polymorphism (RFLP) assay was used to detect and identify most of the species-specific Mycoplasmas involved in cell cultures and virus stock contaminants. Infected cultures detected by PCR-RFLP were further treated with BM-cyclin (5 microg/mL) and passaged for three times and tested for Mycoplasma infections by PCR-RFLP. RESULTS: Mycoplasma pirum and Mycoplasma orale infections were detected by nested PCR. Species specificity was identified by using RFLP of Vsp I, Cla I and Hin dIII restriction enzymes. Mycoplasma infections were cured by treatment with BM-cyclin. This was further confirmed by non-amplification of PCR amplimers in BM-cyclin treated vs. non-treated cultures. CONCLUSIONS: Regular monitoring of cell cultures for Mycoplasma infections and identification of species-specific Mollicutes will identify the source of contaminations. This approach can be used for quality control of the biological reagents used in cell culture and virology laboratories.     

Cervical cancer prevention-related knowledge and attitudes among female undergraduate students from different ethnic groups within China,a survey-based study

ABSTRACT

The purpose of this study was to understand cervical cancer prevention-related knowledge and attitudes among female undergraduate students from different ethnic groups within China. We conducted a survey among ethnically diverse female students from the Minzu University of China, in Beijing in October, 2014. Results: Questionnaires from 493 participants aged from 16 to 26 years were included in the final database. The seven ethnic groups included in the final analysis were Han, Korean, Mongolian, Uyghur, Tibetan, Hui, and Tujia. Compared to the Han Chinese, the members of the other six ethnic groups had lower cervical cancer knowledge levels. The knowledge scores of Mongolian and Korean students were significantly lower than those of the Han Chinese. The willingness to accept cervical cancer prevention efforts also differed across different ethnic groups. After adjusting for age and place of residence, the acceptance of cervical cancer screening among the Tibetan, Uyghur, and Korean groups was significantly lower than among the Han Chinese, with different related decision-making factors in each group. Cervical cancer prevention-related public education is an urgent need in China. Extra consideration of ethnic differences should be taken into account when designing and improving new current cervical cancer prevention programs.     

A Novel Way of Standardization of ICG Lymphangiography Reporting

Background  Indocyanine green (ICG) lymphangiography is being increasingly employed to assess the severity of lymphedema, locate the areas of patent linear lymphatics and dermal backflow and plan treatment. This study suggests a novel method of reporting ICG findings in extremities to enable easy understanding among surgeons and physiotherapists and avoid repeat testing when a patient visits a disparate lymphedema center or clinician. Methods  A reporting protocol was developed in the lymphedema clinic of the plastic surgery department, and patients were asked to bring along the report in every subsequent review. The ICG findings were recorded on the fluorescence imaging system as well. The report was prepared by one and analyzed by two different clinicians without repeating the test on 10 consecutive patients. Results  The interrater reliability of findings in the report was found to be 98.7% among the three clinicians. Conclusion  The reporting system was found to be illustratable and reproducible     

Formulation and In Vitro Evaluation of Ofloxacin Tablets using Natural Gums as Binders

Natural gums are economical, easily available, and useful as tablet binders. In the present investigation, an attempt was made to formulate Ofloxacin tablets using three natural binders, namely Acacia arabica, Hibiscus esculentus, and xanthan gum. Such six batches of Ofloxacin tablets were prepared by using different types and amounts of the natural binders by the wet granulation method. The tablets were analyzed for their hardness, friability, and weight variation, and in vitro release was performed in a phosphate buffer at pH 6.8. The prepared tablets were also evaluated for their various release kinetics and similarity factors f₂. The physical properties of the tablets containing the natural binders showed sufficient hardness, desirable disintegration time, and low friability. Their better percentage of drug release was observed as compared to the marketed formulation showing more than 85% drug release within 45 minutes. The in vitro release data was well-fitted into zero-order and the values of release exponent ‘n’ were between 0.303 and 0.514. The high similarity factor f₂ of 64.50 was achieved with the best batch in comparison to the marketed tablets. The results obtained indicated that the gum Acacia arabica performed as well as gelatin compared to the other binders for the Ofloxacin tablet formulation.     

Microscopic Visualization of Metabotropic Glutamate Receptors on the Surface of Living Cells Using Bifunctional Magnetic Resonance Imaging Probes

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