Delivery and expression of plasmid DNA into cells by a novel non-thermal plasma source.

Medical applications such as plasma assisted gene transfer is a minimally invasive approach that can substantially reduce potential discomfort of treated area. Atmospheric pressure plasma discharge is an effective approach to deliver plasmid DNA for in vitro and in vivo applications. We investigated plasma assisted delivery in vitro in mouse melanoma cells (B16F10) using a novel surface plasma device, which is operated in air. We evaluated the influence of applied voltage and distance between the surface device and cell monolayer. We found no significant effect on the viability of cells. Highest expression following delivery of a plasmid encoding green fluorescent protein was achieved with an applied voltage of 11.25 kV at a 2 mm distance and 5 s exposure time. To better understand the influence of oxidative damages and stress on cells after plasma delivery, a mRNA expression study was performed. Our results indicated that TNFα mRNA was significantly upregulated. The mRNA response may be attributed to the RONS generated by plasma; however, this mRNA upregulation was not adequate to be reflected in a coordinate protein upregulation. From the results reported here, it is clear that this novel plasma device could be used for plasmid delivery.

Changing trends in the etiology and management of vesicovaginal fistula.

Vesicovaginal fistula has remained a scourge and of public health importance, causing significant morbidity, and psychological and social problems to the patient. Continuous wetness, odor and discomfort cause serious social issues. The diagnosis has been traditionally based on clinical evaluation, dye testing, cystoscopic examination and contrast studies. A successful repair of such fistulas requires an accurate diagnosis and timely surgical intervention using techniques that are based on basic surgical principles with or without the use of interpositional flaps. The method of repair depends on the type and location of the fistula, and the surgeon's training and expertise. The main complications are recurrence and stress/urge incontinence. Prevention must include universal education, improvement in the social and nutritional status of women, discouraging early marriages, and the provision of improved accessible healthcare services.

Activated air produced by shielded sliding discharge plasma mediates plasmid DNA delivery to mammalian cells.

Cold plasma is emerging as a potential method for medical applications. The current study assessed the efficacy of a novel cold plasma reactor based on shielded sliding discharge producing cathode-directed streamers generated in ambient air for the delivery of plasmid DNA. Experiments were performed with mouse melanoma cells (B16F10) and human keratinocyte cells (HaCaT) inoculated with plasmid DNA encoding luciferase. Quantitative results measured over a 72-h period displayed luciferase expression levels as high as 5-fold greater in cells exposed to plasma-activated air (PAA) than levels obtained from the inoculation of plasmid DNA alone (P < 0.05, P < 0.01). No effect on cell viability was observed. Delivery of plasmid encoding GFP to HaCaT cells seeded on polycaprolactone (PCL) scaffolds was confirmed by immunostaining. The use of cold plasma for DNA delivery is attractive as it provides a non-viral, non-invasive method where the electrode or the plasma itself never directly contacts the exposed site. The current device design provides localized DNA transfer using a novel technology. Our report suggests PAA warrants further exploration as an alternative or supplemental approach for DNA transfer.

A comparison of the Glidescope, Pentax AWS, and Macintosh laryngoscopes when used by novice personnel: a manikin study.

PURPOSE: Direct laryngoscopic tracheal intubation is a potentially lifesaving procedure, but a difficult skill to acquire and maintain. The consequences of poorly performed intubation attempts are potentially severe. The Pentax AWS and the Glidescope are indirect laryngoscopes that may require less skill to use. We therefore hypothesized that AWS and Glidescope would prove superior to the Macintosh laryngoscope when used by novices in the normal and simulated difficult airway. METHODS: In this prospective randomized crossover trial following standardized didactic instruction, medical students with no prior experience of laryngoscopy performed tracheal intubation using each device. Each student was allowed up to three attempts to intubate in a Laerdal Intubation Trainer in two laryngoscopy scenarios and in a Laerdal SimMan manikin in one scenario. The students then performed tracheal intubation of the normal airway a second time to characterize the learning curve for each device. RESULTS: The Pentax AWS provided better intubation conditions than the Glidescope or the Macintosh, resulting in greater success of intubation, particularly in the difficult laryngoscopy scenarios. The Glidescope demonstrated advantages over the Macintosh, particularly in the more difficult scenarios. Both the AWS and the Glidescope decreased the duration of intubation attempts, reduced the number of maneuvers required, and reduced the potential for dental trauma. In direct comparisons, the AWS provided the best intubation conditions. CONCLUSIONS: The Pentax AWS appears to constitute a better alternative to the Macintosh for novice personnel to acquire the skills of tracheal intubation.