Cryopreserved tissue engineered mucosa.

Single cells suspensions used for grafting in the clinical setting may be reliably cryopreserved by established protocols. However, for tissue engineered constructs which now also get used as grafts in the clinic such established protocols and assays which indicate graft viability and their function as graft do not exist. a) The purpose was to develop a cryoprotocol and an animal model to test the efficacy of tissue engineered to act as graft after cryopreservation. b) Therefore, tissue engineered mucosa grafts consisting of keratinocytes and fibroblasts grown in a collagen sponge were cryopreserved and grafted in the nude rat to test its efficacy to function as mucosa graft. At different points after cryopreservation the mucosa was grafted into the nude rats. Healing of grafts was allowed for one or three weeks. c) Sufficient cells survived the cryopreservation allowing for the development of epithelial-fibroblast tissue in the collagen sponge. After three weeks of healing the formation of mucosa tissue was more complete and more collagen sponge had disappeared. d) The nude rat model is suitable to assess the efficacy of tissue engineered mucosa to function as graft after cryopreservation. The formation of human epithelial-fibroblast tissue in vivo has to be interpreted as proof of principle that the approach of cryopreservation of tissue engineered grafts is working

Assay of Vitamin B6 (Pyridoxine Hydrochloride) Utilizing Isocratic Reversed Phase High Performance Liquid Chromatography

Aims: To demonstrate an analysis for vitamin B6 from commercial aqueous nutritional drinks and solid tablets, utilizing isocratic conditions with high performance liquid chromatography (HPLC) and UV detection at 290 nm. Study Design: Vitamin B6 in the form of pyridoxine hydrochloride is assayed by HPLC from various samples. Place and Duration of Study: Department of Chemistry, Durham Science Center, University of Nebraska, Omaha Nebraska from May to August 2016. Methodology: Utilizing a reversed-phase C-18 column with eluent solvent (19% ethanol, 77% water, 4% acetic acid), the samples were prepared in sample solvent (19% ethanol, 81% distilled water). Detection of vitamin B6 was accomplished at 290 nm. Analysis of samples was done following solubilizing in aqueous conditions having ethanol at 10% to 20% (v/v). Column pressure at 1900 psig, rise time 0.1 with flow rate 1.0 mL/minute. Elution peak for vitamin B6 occurred consistently at 1.6 minutes. Nutritional samples, aqueous samples, and solid pills were prepared in aqueous solvent with various levels of ethanol. Results: Levels of vitamin B6 detected were as low as 4.4029x10-5 molar to 7.8081x10-4 molar. Sensitivity for vitamin B6 was highest at 290 nm. Reverse phase isocratic conditions is shown to be effective for determination of vitamin B6 in aqueous based samples. Standard curves applied are highly linear in range from zero to 7.8081x10-4 molar (y = 112,521,145.5x + 2,818.6), having coefficient of determination (R2 = 0.9948) with very strong positive correlation coefficient (r= 0.9974). Percent recovery of vitamin ranged from 95% to 105%. Amounts of vitamin present in drinks from same manufacturer were consistent. Conclusion: Utilizing reversed phase column, isocratic solvent conditions with ethanol in water, and a UV detector set at 290 nm is effective for determination of vitamin B6. Ethanol-water solvent system is effective. Vitamin B6 was found in various amounts in nutritional drinks tested. Percent recovery of vitamin averaged 101% with a standard deviation of 2.4%. Pyridoxine hydrochloride was effectively assayed from aqueous samples, vitamin preparations, and vitamin tablets. The methodology presented in this study will be useful for quality control analysis for commercial production. Analysis methods for vitamin assay are a necessary objective to ensure quality control of commercial products and medicinal applications

A Knotted Meta-molecule with 2-D Isotropic Optical Activity Rotating the Incident Polarization by 90{\deg}

Optical activity is the ability of chiral materials to rotate linearly-polarized (LP) electromagnetic waves. Because of their intrinsic asymmetry, traditional chiral molecules usually lack isotropic performance, or at best only possess a weak form of chirality. Here we introduce a knotted chiral meta-molecule that exhibits optical activity corresponding to a 90{\deg} polarization rotation of the incident waves. More importantly, arising from the continuous multi-fold rotational symmetry of the chiral torus knot structure, the observed polarization rotation behavior is found to be independent of how the incident wave is polarized. In other words, the proposed chiral knot structure possesses two-dimensional (2-D) isotropic optical activity as illustrated in Fig. 1, which has been experimentally validated in the microwave spectrum. The proposed chiral torus knot represents the most optically active meta-molecule reported to date that is intrinsically isotropic to the incident polarization

Effectiveness of Ledipasvir/Sofosbuvir with/without Ribavarin in Liver Transplant Recipients with Hepatitis C.

Background and Aims: Recurrent infection of hepatitis C virus (HCV) in liver transplant (LT) recipients is universal and associated with significant morbidity and mortality. Methods: We retrospectively evaluated the safety and efficacy of ledipasvir/sofosbuvir with and without ribavirin in LT recipients with recurrent genotype 1 hepatitis C. Results: Eighty-five LT recipients were treated for recurrent HCV with ledipasvir/sofosbuvirwith and without ribavirin for 12 or 24 weeks. The mean (± standard deviation [SD]) time from LT to treatment initiation was 68 (±71) months. The mean (± SD) age of the cohort was 63 (±8.6) years old. Most recipients were male (70%). Baseline alanine transaminase, total bilirubin, and HCV ribonucleic acid (RNA) values (± SD) were 76.8 (±126) mg/dL, 0.8 (±1.3) U/L, and 8,010,421.9 (±12,420,985) IU/mL, respectively. Five of 43 recipients who were treated with ribavirin required drug cessation due to side effects, with 4 of those being anemia complications. No recipient discontinued the ledipasvir/sofosbuvir. Eighty-one percent of recipients had undetectable viral levels at 4 weeks after starting therapy, and all recipients had complete viral suppression at the end of therapy. The sustained viral response at 12 weeks after completion of therapy was 94%. Conclusion : Ledipasvir and sofosbuvir with and without ribavirin therapy is an effective and well-tolerated interferon-free treatment for recurrent HCV infection after LT. Anemia is not uncommon in LT recipients receiving ribavirin

Statistical Learning in Chip (SLIC) (Invited Paper)

Abstract-Despite best efforts, integrated systems are "born" (manufactured) with a unique 'personality' that stems from our inability to precisely fabricate their underlying circuits, and create software a priori for controlling the resulting uncertainty. It is possible to use sophisticated test methods to identify the bestperforming systems but this would result in unacceptable yields and correspondingly high costs. The system personality is further shaped by its environment (e.g., temperature, noise and supply voltage) and usage (i.e., the frequency and type of applications executed), and since both can fluctuate over time, so can the system's personality. Systems also "grow old" and degrade due to various wear-out mechanisms (e.g., negative-bias temperature instability), and unexpectedly due to various early-life failure sources. These "nature and nurture" influences make it extremely difficult to design a system that will operate optimally for all possible personalities. To address this challenge, we propose to develop statistical learning in-chip (SLIC). SLIC is a holistic approach to integrated system design based on continuously learning key personality traits on-line, for selfevolving a system to a state that optimizes performance hierarchically across the circuit, platform, and application levels. SLIC will not only optimize integrated-system performance but also reduce costs through yield enhancement since systems that would have before been deemed to have weak personalities (unreliable, faulty, etc.) can now be recovered through the use of SLIC

Standardized Methods for Enhanced Quality and Comparability of Tuberculous Meningitis Studies

Tuberculous meningitis (TBM) remains a major cause of death and disability in tuberculosis-endemic areas, especially in young children and immunocompromised adults. Research aimed at improving outcomes is hampered by poor standardization, which limits study comparison and the generalizability of results. We propose standardized methods for the conduct of TBM clinical research that were drafted at an international tuberculous meningitis research meeting organized by the Oxford University Clinical Research Unit in Vietnam. We propose a core dataset including demographic and clinical information to be collected at study enrollment, important aspects related to patient management and monitoring, and standardized reporting of patient outcomes. The criteria proposed for the conduct of observational and intervention TBM studies should improve the quality of future research outputs, can facilitate multicenter studies and meta-analyses of pooled data, and could provide the foundation for a global TBM data repository

Adjunctive dexamethasone for the treatment of HIV-uninfected adults with tuberculous meningitis stratified by Leukotriene A4 hydrolase genotype (LAST ACT): Study protocol for a randomised double blind placebo controlled non-inferiority trial [version 1; referees: 2 approved]

Background: Tuberculosis kills more people than any other bacterial infection worldwide. In tuberculous meningitis (TBM), a common functional promoter variant (C/T transition) in the gene encoding leukotriene A4 hydrolase (LTA4H), predicts pre-treatment inflammatory phenotype and response to dexamethasone in HIV-uninfected individuals. The primary aim of this study is to determine whether LTA4H genotype determines benefit or harm from adjunctive dexamethasone in HIV-uninfected Vietnamese adults with TBM. The secondary aim is to investigate alternative management strategies in individuals who develop drug induced liver injury (DILI) that will enable the safe continuation of rifampicin and isoniazid therapy.  Methods: We will perform a parallel group, randomised (1:1), double blind, placebo-controlled,  multi-centre Phase III non-inferiority trial, comparing dexamethasone versus placebo for 6-8 weeks in addition to standard anti-tuberculosis treatment in HIV-uninfected patients with TBM stratified by LTA4H genotype. The primary endpoint will be death or new neurological event. The trial will enrol approximately 720 HIV-uninfected adults with a clinical diagnosis of TBM, from two hospitals in Ho Chi Minh City, Vietnam. 640 participants with CC or CT- LTA4H genotype will be randomised to either dexamethasone or placebo, and the remaining TT- genotype participants will be treated with standard-of-care dexamethasone. We will also perform a randomised comparison of three management strategies for anti-tuberculosis DILI. An identical ancillary study will also be perfomed in the linked randomised controlled trial of dexamethasone in HIV-infected adults with TBM (ACT HIV).  Discussion: Previous data have shown that LTA4H genotype may be a critical determinant of inflammation and consequently of adjunctive anti-inflammatory treatment response in TBM. We will stratify dexamethasone therapy according to LTA4H genotype in HIV-uninfected adults, which may indicate a role for targeted anti-inflammatory therapy according to variation in LTA4H C/T transition. A comparison of DILI management strategies may allow the safe continuation of rifampicin and isoniazid

Expression of Nestin by Neural Cells in the Adult Rat and Human Brain

Neurons and glial cells in the developing brain arise from neural progenitor cells (NPCs). Nestin, an intermediate filament protein, is thought to be expressed exclusively by NPCs in the normal brain, and is replaced by the expression of proteins specific for neurons or glia in differentiated cells. Nestin expressing NPCs are found in the adult brain in the subventricular zone (SVZ) of the lateral ventricle and the subgranular zone (SGZ) of the dentate gyrus. While significant attention has been paid to studying NPCs in the SVZ and SGZ in the adult brain, relatively little attention has been paid to determining whether nestin-expressing neural cells (NECs) exist outside of the SVZ and SGZ. We therefore stained sections immunocytochemically from the adult rat and human brain for NECs, observed four distinct classes of these cells, and present here the first comprehensive report on these cells. Class I cells are among the smallest neural cells in the brain and are widely distributed. Class II cells are located in the walls of the aqueduct and third ventricle. Class IV cells are found throughout the forebrain and typically reside immediately adjacent to a neuron. Class III cells are observed only in the basal forebrain and closely related areas such as the hippocampus and corpus striatum. Class III cells resemble neurons structurally and co-express markers associated exclusively with neurons. Cell proliferation experiments demonstrate that Class III cells are not recently born. Instead, these cells appear to be mature neurons in the adult brain that express nestin. Neurons that express nestin are not supposed to exist in the brain at any stage of development. That these unique neurons are found only in brain regions involved in higher order cognitive function suggests that they may be remodeling their cytoskeleton in supporting the neural plasticity required for these functions