Sonoporation-mediated gene transfer into adult rat dorsal root ganglion cells

<p>Abstract</p> <p>Background</p> <p>Gene transfer into many cell types has been successfully used to develop alternative and adjunct approaches to conventional medical treatment. However, effective transfection of postmitotic neurons remains a challenge. The aim of this study was to develop a method for gene transfer into rat primary dorsal root ganglion neurons using sonoporation.</p> <p>Methods</p> <p>Dissociated cells from adult rat dorsal root ganglion (DRG) cells were sonicated for 1-8 s at 2.5-10 W to determine the optimal ultrasound duration and power for gene transfection and cell survival. Transfection efficiency was compared between sonoporation, liposome and lentiviral vector gene transfer techniques.</p> <p>Results</p> <p>The optimum ultrasound intensity was 5 W for 2 s and yielded an efficiency of gene transfection of 31% and a survival rate of 35%.</p> <p>Conclusions</p> <p>Sonoporation can be optimized to minimize cell death and yield a high percentage of transfected neurons and that this technique can be easily applied to primary cultures of rat dorsal root ganglion neurons.</p

Tear secretion dysfunction among women workers engaged in light-on tests in the TFT-LCD industry

BACKGROUND: The TFT-LCD (thin film transistor liquid crystal display) industry is rapidly growing in Taiwan and many other countries. A large number of workers, mainly women, are employed in the light-on test process to detect the defects of products. At the light-on test workstation, the operator is generally exposed to low humidity (in the clean room environment), flashing light, and low ambient illumination for long working hours. Many workers complained about eye discomfort, and therefore we conducted a study to evaluate the tear secretion function of light-on test workers of a TFT-LCD company. METHODS: We recruited workers engaged in light-on tests in the company during their periodical health examination. In addition to a questionnaire survey of demographic characteristics and ophthalmic symptoms, we evaluated the tear secretion function of both eyes of each participant using the Schirmer's lacrimal basal secretion test with anaesthesia. A participant with one or both eyes yielding abnormal test results was defined as a case of tear secretion dysfunction. RESULTS: During the study period, a total of 371 light-on test workers received the health examination at the clinic of the park, and 52 of them were excluded due to having ophthalmic diseases and other systemic diseases that may affect ophthalmic function. All the remaining 319 qualified workers agreed to participate in this study, and they were all females working by 4-shift rotations. The average age was 24.2 years old (standard deviation [SD] = 3.8), and the average employment duration was 13.6 months (SD = 5.7). Among the 11 ophthalmic symptoms evaluated, eye dryness was the most prevalent (prevalence = 43.3%). In addition, the prevalence of tear secretion dysfunction in at least one eye was 40.1% (128 cases), and contact lens users had an odds ratio of 1.73 (95% confidence interval = 1.02–2.94) in comparison with non-contact lens users. Comparing the Schirmer's test results of those who also participated in the screening in the previous year, we found 40 of the 156 participants (17.2%) with normal test results in the previous year turned abnormal in 2001. In contrast, only 21 of the 76 participants (9.1%) with abnormal test results in the previous year turned normal, and the difference was statistically significant (p = 0.02 for McNemar's test). CONCLUSION: The prevalence of tear secretion dysfunction in woman workers engaged in light-on tests is high and increases with a one-year duration of employment. The use of contact lens may further increase the risk

N348I in the Connection Domain of HIV-1 Reverse Transcriptase Confers Zidovudine and Nevirapine Resistance

Analyzing HIV sequences from a Canadian cohort, Gilda Tachedjian and colleagues identify a common mutation in a little-studied domain of reverse transcriptase that confers resistance to two drug classes

Electromagnetic Wave Theory and Applications

Contains table of contents for Section 3, reports on six research projects and a list of publications and conference papers.Joint Services Electronics Program Contract DAAL03-89-C-0001National Science Foundation Grant ECS 86-20029Schlumberger- Doll ResearchU.S. Army Research Office Contract DAAL03 88-K-0057U.S. Navy - Office of Naval Research Contract N00014-90-J-1002National Aeronautics and Space Administration Grant NAGW-1617U.S. Navy - Office of Naval Research Grant N00014-89-J-1107National Aeronautics and Space Administration Grant NAGW-1272National Aeronautics and Space Administration Agreement 958461U.S. Army - Corps of Engineers Contract DACA39-87-K-0022U.S. Air Force - Electronic Systems Division Contract F19628-88-K-0013U.S. Navy - Office of Naval Research Grant N00014-89-J-1019Digital Equipment CorporationIBM CorporationU.S. Department of Transportation Contract DTRS-57-88-C-00078Defence Advanced Research Projects Agency Contract MDA972-90-C-002

Electromagnetic Wave Theory and Applications

Contains table of contents for Section 3 and reports on five research projects.U.S. Department of Transportation Contract DTRS-57-88-C-00078TTD13U.S. Department of Transportation Contract DTRS-57-88-C-00078TTD30Defense Advanced Research Projects Agency Contract MDA972-90-C-0021Digital Equipment CorporationIBM CorporationJoint Services Electronics Program Contract DAAL03-89-C-0001Joint Services Electronics Program Contract DAAL03-92-C-0001Schlumberger-Doll ResearchU.S. Navy - Office of Naval Research Grant N00014-90-J-1002U.S. Navy - Office of Naval Research Grant N00014-89-J-1019National Aeronautics and Space Administration Grant NAGW-1617National Aeronautics and Space Administration Grant 958461National Aeronautics and Space Administration Grant NAGW-1272U.S. Army Corp of Engineers Contract DACA39-87-K-0022U.S. Navy - Office of Naval Research Grant N00014-89-J-110

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

A MEMS µ-Preconcentrator Employing a Carbon Molecular Sieve Membrane for Highly Volatile Organic Compound Sampling

This paper presents the synthesis and evaluation of a carbon molecular sieve membrane (CMSM) grown inside a MEMS-fabricated μ-preconcentrator for sampling highly volatile organic compounds. An array of µ-pillars measuring 100 µm in diameter and 250 µm in height were fabricated inside a microfluidic channel to increase the attaching surface for the CMSM. The surface area of the CMSM was measured as high as 899 m2/g. A GC peak amplification factor &gt;2 × 104 was demonstrated with gaseous ethyl acetate. Up to 1.4 L of gaseous ethanol at the 100 ppb level could be concentrated without exceeding the capacity of this microchip device. Sharp desorption chromatographic peaks (&lt;3.5 s) were obtained while using this device directly as a GC injector. Less volatile compounds such as gaseous toluene, m-xylene, and mesitylene appeared to be adsorbed strongly on CMSM, showing a memory effect. Sampling parameters such as sample volatilities, sampling capacities, and compound residual issues were empirically determined and discussed

Molecular Mechanism by Which the K70E Mutation in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Confers Resistance to Nucleoside Reverse Transcriptase Inhibitors

The K70E mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) has become more prevalent in clinical samples, particularly in isolates derived from patients for whom triple-nucleoside regimens that include tenofovir (TNV), abacavir, and lamivudine (3TC) failed. To elucidate the molecular mechanism by which this mutation confers resistance to these nucleoside RT inhibitors (NRTI), we conducted detailed biochemical analyses comparing wild-type (WT), K70E, and K65R HIV-1 RT. Pre-steady-state kinetic experiments demonstrate that the K70E mutation in HIV-1 RT allows the enzyme to discriminate between the natural deoxynucleoside triphosphate substrate and the NRTI triphosphate (NRTI-TP). Compared to the WT enzyme, K70E RT showed 2.1-, 2.3-, and 3.5-fold-higher levels of resistance toward TNV-diphosphate, carbovir-TP, and 3TC-TP, respectively. By comparison, K65R RT demonstrated 12.4-, 12.0-, and 13.1-fold-higher levels of resistance, respectively, toward the same analogs. NRTI-TP discrimination by the K70E (and K65R) mutation was primarily due to decreased rates of NRTI-TP incorporation and not to changes in analog binding affinity. The K65R and K70E mutations also profoundly impaired the ability of RT to excise 3′-azido-2′,3′-dideoxythymidine monophosphate (AZT-MP) and other NRTI-MP from the 3′ end of a chain-terminated primer. When introduced into an enzyme with the thymidine analog mutations (TAMs) M41L, L210W, and T215Y, the K70E mutation inhibited ATP-mediated excision of AZT-MP. Taken together, these findings indicate that the K70E mutation, like the K65R mutation, reduces susceptibility to NRTI by selectively decreasing NRTI-TP incorporation and is antagonistic to TAM-mediated nucleotide excision