Electroporation enhances reporter gene expression following delivery of naked plasmid DNA to the lung

BACKGROUND: Existing methods of non-viral airway gene transfer suffer from low levels of efficiency. Electroporation has been used to enhance gene transfer in a range of tissues. Here we assess the usefulness of electroporation for enhancing gene transfer in the lungs of mice and sheep. METHODS: Naked plasmid DNA (pDNA) expressing either luciferase or green fluorescent protein (GFP) was delivered to mouse lungs by instillation. Following surgical visualisation, the lungs were directly electroporated and the level and duration of luciferase activity was assessed and cell types that were positive for GFP were identified in lung cryosections. Naked pDNA was nebulised to the sheep lung and electrodes attached to the tip of a bronchoscope were used to electroporate airway segment bifurcations, Luciferase activity was assessed in electroporated and control non-electroporated regions, after 24 h. RESULTS: Following delivery of naked pDNA to the mouse lung, electroporation resulted in up to 400-fold higher luciferase activity than naked pDNA alone when luciferase was under the control of a cytomegalovirus (CMV) promoter. Following delivery of a plasmid containing the human polyubiquitin C (UbC) promoter, electroporation resulted in elevated luciferase activity for at least 28 days. Visualisation of GFP indicated that electroporation resulted in increased GFP detection compared with non-electroporated controls. In the sheep lung electroporation of defined sites in the airways resulted in luciferase activity 100-fold greater than naked pDNA alone. CONCLUSIONS: These results indicate that electroporation can be used to enhance gene transfer in the lungs of mice and sheep without compromising the duration of expression.

CT radiation dose for computer-assisted endoscopic sinus surgery: dose survey and determination of dose-reduction limits

BACKGROUND AND PURPOSE: Computer-assisted navigation is increasingly used in functional endoscopic sinus surgery (FESS) to prevent injury to vital structures, necessitating preparative CT and, thus, radiation exposure. The purpose of our study was to investigate currently used radiation doses for CT in computer-assisted navigation in sinus surgery (CAS-CT) and to assess minimal doses required. MATERIALS AND METHODS: A questionnaire inquiring about dose parameters used for CAS-CT was sent to 30 radiologic institutions. The feasibility of low-dose registration was tested with a phantom. The influence of CAS-CT dose on technical accuracy and on the practical performance of 5 ear, nose, and throat (ENT) surgeons was evaluated with cadaver heads. RESULTS: The questionnaire response rate was 63%. Variation between minimal and maximal dose used for CAS-CT was 18-fold. Phantom registration was possible with doses as low as 1.1 mGy. No dose dependence on technical accuracy was found. ENT surgeons were able to identify anatomic landmarks on scans with a dose as low as 3.1 mGy. CONCLUSIONS: The vast dose difference between institutions mirrors different attitudes toward image quality and radiation-protection issues rather than being technically founded, and many patients undergo CAS-CT at higher doses than necessary. The only limit for dose reduction in CT for computer-assisted endoscopic sinus surgery is the ENT surgeon's ability to cope with impaired image quality, whereas there is no technically justified lower dose limit. We recommend, generally, doses used for the typical diagnostic low-dose sinus CT (120 kV/20-50 mAs). When no diagnostic image quality is needed, even a reduction down to a third is possible.

Measuring the elastic modulus of polymers using the atomic force microscope

Testing a new method of nanoindentation using the atomic force microscope (AFM) was the purpose of this research. Nanoindentation is a useful technique to study the properties of materials on the sub-micron scale. The AFM has been used as a nanoindenter previously; however several parameters needed to obtain accurate results, including tip radius and cantilever sensitivity, can be difficult to determine. To solve this problem, a new method to determine the elastic modulus of a material using the atomic force microscope (AFM) has been proposed by Tang et al. This method models the cantilever and the sample as two springs in a series. The ratio of the cantilever spring constant (k) to diameter of the tip (2a) is treated in the model as one parameter (α=k/2a). The value of a, along with the cantilever sensitivity, are determined on two reference samples with known mechanical properties and then used to find the elastic modulus of an unknown sample. To determine the reliability and accuracy of this technique, it was tested on several polymers. Traditional depth-sensing nanoindentation was preformed for comparison. The elastic modulus values from the AFM were shown to be statistically similar to the nanoindenter results for three of the five samples tested.

High precision range estimation techniques for multi-path wireless environments

Range estimation is the core of many positioning systems such as radar, and Wireless Local Positioning Systems (WLPS). The estimation of range is achieved by estimating Time-of-Arrival (TOA). TOA represents the signal propagation delay between a transmitter and a receiver. Thus, error in TOA estimation causes degradation in range estimation performance. In wireless environments, noise, multipath, and limited bandwidth reduce TOA estimation performance. TOA estimation algorithms that are designed for wireless environments aim to improve the TOA estimation performance by mitigating the effect of closely spaced paths in practical (positive) signal-to-noise ratio (SNR) regions. Limited bandwidth avoids the discrimination of closely spaced paths. This reduces TOA estimation performance. TOA estimation methods are evaluated as a function of SNR, bandwidth, and the number of reflections in multipath wireless environments, as well as their complexity. In this research, a TOA estimation technique based on Blind signal Separation (BSS) is proposed. This frequency domain method estimates TOA in wireless multipath environments for a given signal bandwidth. The structure of the proposed technique is presented and its complexity and performance are theoretically evaluated. It is depicted that the proposed method is not sensitive to SNR, number of reflections, and bandwidth. In general, as bandwidth increases, TOA estimation performance improves. However, spectrum is the most valuable resource in wireless systems and usually a large portion of spectrum to support high performance TOA estimation is not available. In addition, the radio frequency (RF) components of wideband systems suffer from high cost and complexity. Thus, a novel, multiband positioning structure is proposed. The proposed technique uses the available (non-contiguous) bands to support high performance TOA estimation. This system incorporates the capabilities of cognitive radio (CR) systems to sense the available spectrum (also called white spaces) and to incorporate white spaces for high-performance localization. First, contiguous bands that are divided into several non-equal, narrow sub-bands that possess the same SNR are concatenated to attain an accuracy corresponding to the equivalent full band. Two radio architectures are proposed and investigated: the signal is transmitted over available spectrum either simultaneously (parallel concatenation) or sequentially (serial concatenation). Low complexity radio designs that handle the concatenation process sequentially and in parallel are introduced. Different TOA estimation algorithms that are applicable to multiband scenarios are studied and their performance is theoretically evaluated and compared to simulations. Next, the results are extended to non-contiguous, non-equal sub-bands with the same SNR. These are more realistic assumptions in practical systems. The performance and complexity of the proposed technique is investigated as well. This study’s results show that selecting bandwidth, center frequency, and SNR levels for each sub-band can adapt positioning accuracy.

Temporary neurological dysfunction after surgery of the thoracic aorta: a predictor of poor outcome and impaired quality of life

BACKGROUND: Transient neurological dysfunction (TND) consists of postoperative confusion, delirium and agitation. It is underestimated after surgery on the thoracic aorta and its influence on long-term quality of life (QoL) has not yet been studied. This study aimed to assess the influence of TND on short- and long-term outcome following surgery of the ascending aorta and proximal arch. METHODS: Nine hundred and seven patients undergoing surgery of the ascending aorta and the proximal aortic arch at our institution were included. Two hundred and ninety patients (31.9%) underwent surgery because of acute aortic dissection type A (AADA) and 617 patients because of aortic aneurysm. In 547 patients (60.3%) the distal anastomosis was performed using deep hypothermic circulatory arrest (DHCA). TND was defined as a Glasgow coma scale (GCS) value <13. All surviving patients had a clinical follow up and QoL was assessed with an SF-36 questionnaire. RESULTS: Overall in-hospital mortality was 8.3%. TND occurred in 89 patients (9.8%). As compared to patients without TND, those who suffered from TND were older (66.4 vs 59.9 years, p<0.01) underwent more frequently emergent procedures (53% vs 32%, p<0.05) and surgery under DHCA (84.3% vs 57.7%, p<0.05). However, duration of DHCA and extent of surgery did not influence the incidence of TND. In-hospital mortality in the group of patients with TND compared to the group without TND was similar (12.0% vs 11.4%; p=ns). Patients with TND suffered more frequently from coronary artery disease (28% vs 20.8%, p=ns) and were more frequently admitted in a compromised haemodynamic condition (23.6% vs 9.9%, p<0.05). Postoperative course revealed more pulmonary complications such as prolonged mechanical ventilation. Additional to their transient neurological dysfunction, significantly more patients had strokes with permanent neurological loss of function (14.6% vs 4.8%, p<0.05) compared to the patients without TND. ICU and hospital stay were significantly prolonged in TND patients (18+/-13 days vs 12+/-7 days, p<0.05). Over a mean follow-up interval of 27+/-14 months, patients with TND showed a significantly impaired QoL. CONCLUSION: The neurological outcome following surgery of the ascending aorta and proximal aortic arch is of paramount importance. The impact of TND on short- and long-term outcome is underestimated and negatively affects the short- and long-term outcome.

The Calcite Veins of the Livingston Formation

An interesting group of calcite veins occur near Livingston Montana in a zone about eight miles wide and forty miles long in the edge of the Plains region of Montana in front of the Main ranges. The zone extends from the Boulder River south of Big Timber, through Springdale and Hunters Hot Springs, to Potter's Basin just north of Wilsall in Park County. The group of veins is particularly interesting because they cut relatively flat lying strata, suggest a structural relation­ship to one another, and they are nearly pure calcite. The present investigation was to determine the position of the calcite veins by plotting them on a base map of the district. In addition, it was planned to determine the mineralogy end origin of the veins and their structural and stratigraphical relationship to the rocks of the Livingston formation which they cut.

Warranty vs. Non-Warranty Road Options

Comparing the cost effectiveness of warranty vs. non-warranty road construction options for a 100 mile section of road over a 20 year period.

The Acropolitan - v. 5, no. 4

In this issue...Debate Team, K. C. Gym, Constitution, John F. Rickards, Coach McAuliffe, Mines Basketball, Bureau of Mines, Butte, Montana

The Amplifier - v. 3, no. 3

In this issue...Engineering Day, M-Club, Gorden Ziesing, S&H Green Stamps,Shell Oil Company, Dave Johnson, school honor system, B. W. Brown, Stanolind

Innate and adaptive immunity cooperate flexibly to maintain host-microbiota mutualism

Commensal bacteria in the lower intestine of mammals are 10 times as numerous as the body's cells. We investigated the relative importance of different immune mechanisms in limiting the spread of the intestinal microbiota. Here, we reveal a flexible continuum between innate and adaptive immune function in containing commensal microbes. Mice deficient in critical innate immune functions such as Toll-like receptor signaling or oxidative burst production spontaneously produce high-titer serum antibodies against their commensal microbiota. These antibody responses are functionally essential to maintain host-commensal mutualism in vivo in the face of innate immune deficiency. Spontaneous hyper-activation of adaptive immunity against the intestinal microbiota, secondary to innate immune deficiency, may clarify the underlying mechanisms of inflammatory diseases where immune dysfunction is implicated.

Political Debates Strengthen Our Democracy

A Montana Public Radio Commentary by Evan Barrett. Published newspaper columns written by Evan Barrett on this topic, which vary somewhat in content from this commentary, appeared in the following publications: Billings Gazette, September 3, 2014 Montana Standard, September 5, 2014 Missoulian, September 5, 2014 Independent Record, September 7, 2014