A preliminary report on stem cell therapy for neuropathic pain in humans

E Russell Vickers,1 Elisabeth Karsten,2 John Flood,3 Richard Lilischkis21Sydney Oral and Maxillofacial Surgery, NSW, Australia; 2Regeneus Ltd, Gordon, NSW, Australia; 3St Vincents Hospital, Sydney, NSW, AustraliaObjective: Mesenchymal stem cells (MSCs) have been shown in animal models to attenuate chronic neuropathic pain. This preliminary study investigated if: i) injections of autologous MSCs can reduce human neuropathic pain and ii) evaluate the safety of the procedure.Methods: Ten subjects with symptoms of neuropathic trigeminal pain underwent liposuction. The lipoaspirate was digested with collagenase and washed with saline three times. Following centrifugation, the stromal vascular fraction was resuspended in saline, and then transferred to syringes for local injections into the pain fields. Outcome measures at 6 months assessed reduction in: i) pain intensity measured by standard numerical rating scale from 0–10 and ii) daily dosage requirements of antineuropathic pain medication.Results: Subjects were all female (mean age 55.3 years ± standard deviation [SD] 14.67; range 27–80 years) with pain symptoms lasting from 4 months to 6 years and 5 months. Lipoaspirate collection ranged from 102–214 g with total cell numbers injected from 33 million to 162 million cells. Cell viability was 62%–91%. There were no systemic or local tissue side effects from the stem cell therapy (n=41 oral and facial injection sites). Clinical pain outcomes showed that at 6 months, 5/9 subjects had reduced both pain intensity scores and use of antineuropathic medication. The mean pain score pre-treatment was 7.5 (SD 1.58) and at 6 months had decreased to 4.3 (SD 3.28), P=0.018, Wilcoxon signed-rank test. Antineuropathic pain medication use showed 5/9 subjects reduced their need for medication (gabapentin, P=0.053, Student's t-test).Conclusion: This preliminary open-labeled study showed autologous administration of stem cells for neuropathic trigeminal pain significantly reduced pain intensity at 6 months and is a safe and well tolerated intervention.Keywords: adipose, stem cells, neuropathic, orofacial, trigemina

Vertical ozone distribution in the marine atmsophere over the central Atlantic Ocean (56 S - 50 N)

Abstract. The vertical ozone distribution over the Atlantic Ocean has been measured in situ by shipborne ozone soundings during three RV Polarstern meridional transects in January/February 1993, October/November 1993, and May/June 1994. The low ozone column densities measured by Nimbus 7 and Meteor 3 satellites in 1993 could be confirmed by our investigations. We observed distinct differences in the vertical distribution pattern of tropospheric ozone between the northern and the southern hemisphere: The ozone mixing ratio gradients were flat in the northern hemisphere and ozone mixing ratios in the free troposphere never did exceed 80 parts per billion by volume (ppbv) up to the tropopause, while the southern hemisphere exhibited a pronounced vertical gradient. Extremely dry air masses with enhanced ozone amounts up to 120 ppbv have been found in the tropical free troposphere of the southern hemisphere between 0° and 20°S. The vertical ozone stratification in the troposphere of the southern hemisphere was dominated by this large-scale feature. Photochemical ozone production as a consequence of the emissions of natural fires or intrusions of stratospheric air masses are the most probable sources for these ozone-rich layers. On the basis of our results, a stringent differentiation between both alternatives could not be given

Scalable fabrication and application of nanoscale IDE-arrays as multi-electrode platform for label-free biosensing

© 2018 Elsevier B.V. The continuous progress in the construction of advanced, miniaturized electrodes provides a promising route towards compact and sensitive biological and chemical sensor platforms. We present a combined micro- and nanofabrication process at wafer-scale with nanoimprint lithography and subsequent photolithography for the realization of ultra-small, interdigitated electrode arrays. Several chips of gold nanoelectrode arrays (NEA) in a 4 × 4 configuration designed as interdigitated electrodes (NEA-IDEs) with finger structures measuring 14 μm in length and 600 nm in width with 600 nm spacing were fabricated simultaneously on 4-inch wafers. Our process involved a nanoimprint lithography step, wet-etching, metal evaporation and nano lift-off followed by optical lithography for metal contact lines and passivation layers. The optimized procedure yielded high-quality NEA-IDEs with reliable electrochemical behavior as inferred from voltammetric and impedimetric analysis. The final array allows the control of all 16 NEA-IDEs in parallel, which can be beneficial for multi-analyte detection. In a proof-of-concept assay, to demonstrate the applicability of the NEA-IDEs for biosensing, the nanostructures were modified with short DNA molecules as recognition elements for the detection of hybridization via impedance spectroscopy. Stable impedance signals were found using the redox system ferri-/ferrocyanide. After hybridization with complementary target DNA the sensors showed an enhancement of the charge transfer resistance. Experiments with different target DNA concentrations demonstrated a dynamic detection range of 1–100 nM. The main advantage of these NEA-IDE structures is that they are small enough to be integrated into typical microchannel dimensions of 50–100 μm for miniaturized lab-on-a-chip biosensor devices in future.status: publishe

Scalable fabrication and application of nanoscale IDE-arrays as multi-electrode platform for label-free biosensing

International audienceThe continuous progress in the construction of advanced, miniaturized electrodes provides a promising route towards compact and sensitive biological and chemical sensor platforms. We present a combined micro- and nanofabrication process at wafer-scale with nanoimprint lithography and subsequent photolithography for the realization of ultra-small, interdigitated electrode arrays. Several chips of gold nanoelectrode arrays (NEA) in a 4 x 4 configuration designed as interdigitated electrodes (NEA-IDES) with finger structures measuring 14 mu m in length and 600 nm in width with 600 nm spacing were fabricated simultaneously on 4-inch wafers. Our process involved a nanoimprint lithography step, wet-etching, metal evaporation and nano lift-off followed by optical lithography for metal contact lines and passivation layers. The optimized procedure yielded high-quality NEA-IDES with reliable electrochemical behavior as inferred from voltammetric and impedimetric analysis. The final array allows the control of all 16 NEA-IDES in parallel, which can be beneficial for multi-analyte detection. In a proof-of-concept assay, to demonstrate the applicability of the NEA-IDES for biosensing, the nanostructures were modified with short DNA molecules as recognition elements for the detection of hybridization via impedance spectroscopy. Stable impedance signals were found using the redox system ferri-/ferrocyanide. After hybridization with complementary target DNA the sensors showed an enhancement of the charge transfer resistance. Experiments with different target DNA concentrations demonstrated a dynamic detection range of 1-100 nM. The main advantage of these NEA-IDE structures is that they are small enough to be integrated into typical microchannel dimensions of 50-100 mu m for miniaturized lab-on-a-chip biosensor devices in future. (C) 2018 Elsevier B.V. All rights reserved

Mad1 function in cell proliferation and transcriptional repression is antagonized by cyclin E/CDK2

The transcription factors of the Myc/Max/Mad network play essential roles in the regulation of cellular behavior. Mad1 inhibits cell proliferation by recruiting an mSin3-corepressor complex that contains histone deacetylase activity. Here we demonstrate that Mad1 is a potent inhibitor of the G1 to S phase transition, a function that requires Mad1 to heterodimerize with Max and to bind to the corepressor complex. Cyclin E/CDK2, but not cyclin D and cyclin A complexes, fully restored S phase progression. In addition inhibition of colony formation and gene repression by Mad1 were also efficiently antagonized by cyclin E/CDK2. This was the result of cyclin E/CDK2 interfering with the interaction of Mad1 with HDAC1 and reducing HDAC activity. Our findings define a novel interplay between the cell cycle regulator cyclin E/CDK2 and Mad1 and its associated repressor complex and suggests an additional mechanism how cyclin E/CDK2 affects the G1 to S phase transition

Tissue-Specific DNA Methylation Patterns in Forensic Samples Detected by Pyrosequencing®

In certain circumstances the outcome of a trial may hinge on the ability of a forensic laboratory to determine the identity of biological stains present at crime scenes. An example of such a situation would be the detection of blood, saliva, vaginal fluid, or other body fluid in a specific location whereby its presence would reinforce the victim’s or suspect’s version of the events that happened during the commission of a crime. However, current serological methods used for identifying body fluids may lack the sensitivity and specificity to identify these fluids, particularly for trace levels. New procedures using proteomic methods and RNA-based gene expression show promise in addressing this issue; however, concerns about stability and relative levels of gene expression remain. An alternative approach is to utilize patterns of epigenetic DNA methylation. DNA methylation is an epigenetic mechanism that regulates the specificity of genes being expressed or silenced in cells. Regions in the human genome referred to as tissue-specific differentially methylated regions account for unique patterns of DNA methylation that are specific for each cell type. This chapter addresses the application of bisulfite-modified PCR combined with Pyrosequencing® to detect tissue-specific DNA methylation patterns and perform trace serological analysis. The quantitative nature and precision available with Pyrosequencing presents major advantages in these studies as it permits detection of and contrast between cells with differential levels of methylation. The procedure can be applied to a variety of biological fluids which may be present at crime scenes.https://nsuworks.nova.edu/cnso_bio_facbooks/1016/thumbnail.jp