News Article | April 13, 2016
All non-transgenic, transgenic and control mice used in this study were derived from in house breeding colonies backcrossed > 12 generations onto C57/BL6 backgrounds. All mice used were young adult females between two and four months old at the time of spinal cord injury. All transgenic mice used have been previously well characterized or are the progeny of crossing well-characterized lines: (1) mGFAP-TK transgenic mice line 7.115, 16, 49; (2) mGFAP-Cre-STAT3-loxP mice generated by crossing STAT3-loxP mice with loxP sites flanking exon 22 of the STAT3 gene50 with mGFAP-Cre mice line 73.1217, 18; (3) loxP-STOP-loxP-DTR (diphtheria toxin receptor) mice21; (4) mGFAP-Cre-RiboTag mice generated by crossing mice with loxP-STOP-loxP-Rpl22-HA (RiboTag)26 with mGFAP-Cre mice line 73.1217, 18; (5) loxP-STOP-loxP-tdTomato reporter mice51. All mice were housed in a 12-h light/dark cycle in a specific-pathogen-free facility with controlled temperature and humidity and were allowed free access to food and water. All experiments were conducted according to protocols approved by the Animal Research Committee of the Office for Protection of Research Subjects at University of California, Los Angeles. All surgeries were performed under general anaesthesia with isoflurane in oxygen-enriched air using an operating microscope (Zeiss, Oberkochen, Germany), and rodent stereotaxic apparatus (David Kopf, Tujunga, CA). Laminectomy of a single vertebra was performed and severe crush spinal cord injuries (SCI) were made at the level of T10 using No. 5 Dumont forceps (Fine Science Tools, Foster City, CA) without spacers and with a tip width of 0.5 mm to completely compress the entire spinal cord laterally from both sides for 5 s16, 17, 18. For pre-conditioning lesions, sciatic nerves were transected and ligated one week before SCI. Hydrogels were injected stereotaxically into the centre of SCI lesions 0.6 mm below the surface at 0.2 μl per minute using glass micropipettes (ground to 50–100 μm tips) connected via high-pressure tubing (Kopf) to 10-μl syringes under control of microinfusion pumps, two days after SCI52. Tract tracing was performed by injection of biotinylated dextran amine 10,000 (BDA, Invitrogen) 10% wt/vol in sterile saline injected 4 × 0.4 μl into the left motor cerebral cortex 14 days before perfusion to visualize corticospinal tract (CST) axons, or choleratoxin B (CTB) (List Biological Laboratory, Campbell, CA) 1 μl of 1% wt/vol in sterile water injected into both sciatic nerves three days before perfusion to visualize ascending sensory tract (AST) axons33. AAV2/5-GfaABC1D-Cre (see below) was injected either 3 or 6 × 0.4 μl (1.29 × 1013 gc ml−1 in sterile saline) into and on either side of mature SCI lesions two weeks after SCI, or into uninjured spinal cord after T10 laminectomy. All animals received analgesic before wound closure and every 12 h for at least 48 h post-injury. Animals were randomly assigned numbers and evaluated thereafter blind to genotype and experimental condition. Adeno-associated virus 2/5 (AAV) vector with a minimal GFAP promoter (AAV2/5 GfaABC1D) was used to target Cre-recombinase expression selectively to astrocytes53, 54, 55. Diblock co-polypeptide hydrogel (DCH) K L was fabricated, tagged with blue fluorescent dye (AMCA-X) and loaded with growth factor and antibody cargoes as described38, 39, 52. Cargo molecules comprised: human recombinant NT3 and BDNF were gifts (Amgen, Thousand Oaks, CA, (NT3 Lot#2200F4; BDNF Lot#2142F5A) or were purchased from PeproTech (Rocky Hill, NJ; NT3 405-03, Lot#060762; BDNF 405-02 Lot#071161). Function blocking anti-CD29 mouse monoclonal antibody was purchased from BD Bioscience (San Diego, CA) as a custom order at 10.25 mg ml−1 (product #BP555003; lot#S03146). Freeze dried K L powder was reconstituted on to 3.0% or 3.5% wt/vol basis in sterile PBS without cargo or with combinations of NT3 (1.0 μg μl−1), BDNF (0.85 μg μl−1) and anti-CD29 (5 μg μl−1). DCH mixtures were prepared to have G′ (storage modulus at 1 Hz) between 75 and 100 Pascal (Pa), somewhat below that of mouse brain at 200 Pa (refs 38, 39). GCV (Cytovene-IV Hoffman LaRoche, Nutley, NJ), 25 mg kg−1 per day dissolved in sterile physiological saline was administered as single daily subcutaneous injections starting immediately after surgery and continued for the first 7 days after SCI. Bromodeoxyuridine (BrdU, Sigma), 100 mg kg−1 per day dissolved in saline plus 0.007 M NaOH, was administered as single daily intraperitoneal injections on days 2 through 7 after SCI. Diphtheria toxin A (DT, Sigma #DO564) 100 ng in 100 μl sterile saline was administered twice daily as intraperitoneal injections for ten days starting three weeks after injection of AAV2/5-GfaABC1D-Cre to loxP-DTR mice (which was 5 weeks after SCI) (see timeline in Extended Data Fig. 1d). Two days after SCI, all mice were evaluated in open field and mice exhibiting any hindlimb movements were not studied further. Mice that passed this pre-determined inclusion criterion were randomized into experimental groups for further treatments and were thereafter evaluated blind to their experimental condition. At 3, 7, 14 days and then weekly after SCI, hindlimb movements were scored using a simple six-point scale in which 0 is no movement and 5 is normal walking17. After terminal anaesthesia by barbiturate overdose mice were perfused transcardially with 10% formalin (Sigma). Spinal cords were removed, post-fixed overnight, and cryoprotected in buffered 30% sucrose for 48 h. Frozen sections (30 μm horizontal) were prepared using a cryostat microtome (Leica) and processed for immunofluorescence as described16, 17, 18. Primary antibodies were: rabbit anti-GFAP (1:1,000; Dako, Carpinteria, CA); rat anti-GFAP (1:1,000, Zymed Laboratories); goat anti-CTB (1:1,000, List Biology Lab); rabbit anti-5HT (1:2,000, Immunostar); goat anti-5HT (1:1,000, Immunostar); mouse anti-CSPG22 (1:100, Sigma); rabbit-anti haemagglutinin (HA) (1:500 Sigma); mouse-anti HA (1:3,000 Covance); sheep anti-BrdU (1:6,000, Maine Biotechnology Services, Portland, ME); rabbit anti-laminin (1:80, Sigma, Saint Louis, MO); guinea pig anti-NG2 (CSPG4) (E. G. Hughes and D. W. Bergles56, Baltimore, MA); goat anti-aggrecan (1:200, NOVUS); rabbit anti-brevican (1:300, NOVUS); mouse anti-neurocan (1:300, Milipore); mouse anti-phosphacan (1:500, Sigma); goat anti-versican (1:200, NOVUS); rabbit anti-neurglycan C (CSPG5) (1:200, NOVUS). Fluorescence secondary antibodies were conjugated to: Alexa 488 (green) or Alexa 350 (blue) (Molecular Probes), or to Cy3 (550, red) or Cy5 (649, far red) all from (Jackson Immunoresearch Laboratories). Mouse primary antibodies were visualized using the Mouse-on-Mouse detection kit (M.O.M., Vector). BDA tract-tracing was visualized with streptavidin-HRP plus TSB Fluorescein green or Tyr-Cy3 (Jackson Immunoresearch Laboratories). Nuclear stain: 4′,6′-diamidino-2-phenylindole dihydrochloride (DAPI; 2 ng ml−1; Molecular Probes). Sections were coverslipped using ProLong Gold anti-fade reagent (InVitrogen, Grand Island, NY). Sections were examined and photographed using deconvolution fluorescence microscopy and scanning confocal laser microscopy (Zeiss, Oberkochen, Germany). Axons labelled by tract tracing or immunohistochemistry were quantified using image analysis software (NeuroLucida, MicroBrightField, Williston, VT) operating a computer-driven microscope regulated in the x, y and z axes (Zeiss) by observers blind to experimental conditions. Using NeuroLucida, lines were drawn across horizontal spinal cord sections at SCI lesion centres and at regular distances on either side (Fig. 1a) and the number of axons intercepting lines was counted at 63× magnification under oil immersion by observers blind to experimental conditions. Similar lines were drawn and axons counted in intact axon tracts 3 mm proximal to SCI lesions and the numbers of axon intercepts in or near lesions were expressed as percentages of axons in the intact tracts in order to control for potential variations in tract-tracing efficacy or intensity of immunohistochemistry among animals. Two sections at the level of the CST or AST, and three sections through the middle of the cord for 5HT, were counted per mouse and expressed as total intercepts per location per mouse. To determine efficacy of axon transection after SCI, we examined labelling 3 mm distal to SCI lesion centres, with the intention of eliminating mice that had labelled axons at this location on grounds that these mice may have had incomplete lesions. However, all mice that had met the strict behavioural inclusion criterion of no hindlimb movements two days after severe crush SCI, exhibited no detectable axons 3 mm distal to SCI lesions regardless of treatment group. Sections stained for GFAP, CSPG or laminin were photographed using constant exposure settings. Single-channel immunofluorescence images were converted to black and white and thresholded (Fig. 1d and Extended Data Fig. 2b) and the amount of stained area measured in different tissue compartments using NIH ImageJ software. Areas are shown in graphs as mean values plus or minus standard error of the means (s.e.m.). Statistical evaluations of repeated measures were conducted by ANOVA with post hoc, independent pairwise analysis as per Newman-Keuls (Prism, GraphPad, San Diego, CA). Power calculations were performed using G*Power Software v220.127.116.11 (ref. 57). For quantification of histologically derived neuroanatomical outcomes such as numbers of axons or percentage of area stained for GFAP or CSPG, group sizes were used that were calculated to provide at least 80% power when using the following parameters: probability of type I error (α) = 0.05, a conservative effect size of 0.25, 2–8 treatment groups with multiple measurements obtained per replicate. Using Fig. 5j as an example, evaluation of n = 5 biological replicates (with multiple measurements per replicate) in each of 8 treatment groups provided greater than 88% power. For dot blot immunoassay of chondroitin sulfate proteoglycans (CSPG), spinal cord tissue blocks were lysed and homogenized in standard RIPA (radio-immunoprecipitation assay) buffer. LDS (lithium dodecyl sulfate) buffer (Life Technologies) was added to the post-mitochondrial supernatant and 2 μl containing 2 μg μl−1 protein was spotted onto a nitrocellulose membrane (Life Technologies), set to dry and incubated overnight with mouse anti-chondroitin sulfate antibody (CS56, 1:1000, Sigma Aldrich), an IgM-monoclonal antibody that detects glyco-moieties of all CSPGs22. CS56 immunoreactivity was detected on X-ray film with alkaline phosphatase-conjugated secondary antibody and chemiluminescent substrate (Life Technologies). Densitometry measurements of CS56 immunoreactivity were obtained using ImageJ software (NIH) and normalized to total protein (Poncau S) density58. Densities are shown in graphs as mean values plus or minus standard error of the means (s.e.m.). Two weeks after SCI, spinal cords of wild-type control (GFAP-RiboTag) and STAT3-CKO (GFAP-STAT3CKO-RiboTag) mice were rapidly dissected out of the spinal canal. The central 3 mm of the lower thoracic lesion including the lesion core and 1 mm rostral and caudal were then rapidly removed and snap frozen in liquid nitrogen. Haemagglutinin (HA) immunoprecipitation (HA-IP) of astrocyte ribosomes and ribosome-associated mRNA (ramRNA) was carried out as described26. The non-precipitated flow-through (FT) from each IP sample was collected for analysis of non-astrocyte total RNA. HA and FT samples underwent on-column DNA digestion using the RNase-Free Dnase Set (Qiagen) and RNA purified with the RNeasy Micro kit (Qiagen). Integrity of the eluted RNA was analysed by a 2100 Bioanalyzer (Agilent) using the RNA Pico chip, mean sample RIN = 8.0 ± 0.95. RNA concentration determined by RiboGreen RNA Assay kit (Life Technologies). cDNA was generated from 5 ng of IP or FT RNA using the Nugen Ovation 2 RNA-Seq Sytstem V2 kit (Nugen). 1 μg of cDNA was fragmented using the Covaris M220. Paired-end libraries for multiplex sequencing were generated from 300 ng of fragmented cDNA using the Apollo 324 automated library preparation system (Wafergen Biosystems) and purified with Agencourt AMPure XP beads (Beckman Coulter). All samples were analysed by an Illumina NextSeq 500 Sequencer (Illumina) using 75-bp paired-end sequencing. Reads were quality controlled using in-house scripts including picard-tools, mapped to the reference mm10 genome using STAR59, and counted using HT-seq60 with mm10 refSeq as reference, and genes were called differentially expressed using edgeR61. Individual gene expression levels in the Fig. 4e histogram are shown as mean FPKM (fragments per kilobase of transcript sequence per million mapped fragments). Additional details of differential expression analysis are described in the legends of Fig. 4 and Extended Data Figs 3 and 4. Raw and normalized data have been deposited in the NCBI Gene Expression Omnibus and are accessible through accession number GSE76097. To ensure the widespread distribution of these datasets, we have created a user-friendly website that enables searching for individual genes of interest https://astrocyte.rnaseq.sofroniewlab.neurobio.ucla.edu.
Shin Y.S.,National Jewish Health |
Takeda K.,National Jewish Health |
Ohnishi H.,National Jewish Health |
Jia Y.,National Jewish Health |
And 8 more authors.
Annals of Allergy, Asthma and Immunology | Year: 2011
Background: Asthma is a chronic airway inflammatory disease that is associated with a large influx of inflammatory cells. Several chemokines and chemokine receptors play critical roles in the development of allergic airway inflammation. Objective: Because polarized human TH2 cells express a functional CXCR3 chemokine receptor, we evaluated the effects of a selective CXCR3 inhibitor in a mouse model of allergic airway disease. Methods: Ovalbumin-specific CD8+ T effector cells were generated from OT-1 mice in the presence of interleukin 2. The activity of a CXCR3 inhibitor was examined in vitro by monitoring Ca2+ influx after receptor ligation. In vivo, the activity was assessed in sensitized and challenged mice by monitoring airway function, inflammatory parameters, including cellular infiltrates and cytokines in the bronchoalveolar lavage fluid. Results: Approximately 40% of CD8+ T effector cells expressed the CXCR3 receptor. In vitro, CXCR3 antagonism reduced Ca2+ influx after receptor engagement. In contrast, the CXCR3 antagonist had little to no effect on airway function or inflammatory parameters despite adequate exposure levels. Conclusions: CXCR3 antagonism did not prevent allergen-induced airway hyperresponsiveness or airway inflammation in a mouse allergy model despite having activity in in vitro functional assays. © 2011 American College of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.
Zelent B.,University of Pennsylvania |
Buettger C.,University of Pennsylvania |
Grimsby J.,Hoffman LaRoche |
Sarabu R.,Hoffman LaRoche |
And 3 more authors.
Biochimica et Biophysica Acta - Proteins and Proteomics | Year: 2012
We investigated how glycerol, urea, glucose and a GKA influence kinetics and stability of wild-type and mutant GK. Glycerol and glucose stabilized GK additively. Glycerol barely affected the TF spectra of all GKs but decreased k cat, glucose S 0.5 and K D values and ATP K M while leaving cooperativity unchanged. Glycerol sensitized all GKs to GKA as shown by TF. Glucose increased TF of GKs without influence of glycerol on the effect. Glycerol and GKA affected kinetics and binding additively. The activation energies for thermal denaturation of GK were a function of glucose with K Ds of 3 and 1 mM without and with glycerol, respectively. High urea denatured wild type GK reversibly at 20 and 60 °C and urea treatment of irreversibly heat denatured GK allowed refolding as demonstrated by TF including glucose response. We concluded: Glycerol stabilizes GK indirectly without changing the folding structure of the apoenzyme, by restructuring the surface water of the protein, whereas glucose stabilizes GK directly by binding to its substrate site and inducing a compact conformation. Glucose or glycerol (alone or combined) is unable to prevent irreversible heat denaturation above 40 °C. However, urea denatures GK reversibly even at 60 °C by binding to the protein backbone and directly interacting with hydrophobic side chains. It prevents irreversible aggregation allowing complete refolding when urea is removed. This study establishes the foundation for exploring numerous instability mutants among the more than 600 variant GKs causing diabetes in animals and humans. © 2012 Elsevier B.V. All rights reserved.
Lakshman K.M.,Boston University |
Kaplan B.,Lahey Clinic Medical Center |
Travison T.G.,Boston University |
Basaria S.,Boston University |
And 5 more authors.
Journal of Clinical Endocrinology and Metabolism | Year: 2010
Background: During testosterone (T) therapy, T is partly converted to 17β-estradiol (E 2) and 5α-dihydrotestosterone (DHT). Effects of age, testosterone dose, and body composition on total and free E 2 and DHT levels are unknown. Objective: We evaluated age and dose-related differences in E 2 and DHT levels in response to graded doses of testosterone enanthate in young and older men. Methods: Fifty-one young (aged 19-35 yr) and 52 older (aged 59-75 yr) men completed treatment with monthly injections of a GnRH agonist plus randomly assigned weekly doses of testosterone enanthate (25, 50, 125, 300, or 600 mg) for 5 months. Results: During testosterone administration, total and free E 2 levels increased dose-dependently (dose effect, P<0.001) in both young and older men. Total and free E 2 levels and E 2:T ratios during T administration were higher in older than young men, but age-related differences in free E 2 and free E 2:T ratioswerenot significant after adjusting for testosterone levels, percentage fat mass,and SHBG. DHT levels and DHT:T ratios were dose-related but did not differ between young and older men. Mechanistic modeling of free hormone data revealed that the conversions of T to E 2 and DHT were both consistent with saturable Michaelis-Menten kinetics. The in vivo K m values were estimated to be 1.83 nM for aromatase and 3.35 nM for 5α-reductase, independent of age. The V max parameter for E 2 was 40% higher in older men than younger men, but V max for DHT was not significantly different between age groups. Conclusions: During imtestosterone administration, E 2 and DHT levels exhibit saturable increases with dose. The rate of whole body aromatization is higher in older men, partly related to their higher percentage fat mass, SHBG, and testosterone levels. Copyright © 2010 by The Endocrine Society.
Amit O.,Glaxosmithkline |
Mannino F.,Glaxosmithkline |
Stone A.M.,Astrazeneca |
Bushnell W.,Glaxosmithkline |
And 3 more authors.
European Journal of Cancer | Year: 2011
Purpose: Progression free survival (PFS) is increasingly used as a primary end-point in oncology clinical trials. This paper provides observations and recommendations on the use of a blinded independent central review (BICR) for progression. Patients and methods: The findings and recommendations are based on extensive simulations and a meta-analysis based on 27 previously conducted randomised phase III trials with BICR performed by the Pharmaceutical Research and Manufacturers Association (PhRMA) sponsored PFS Independent Review Working Group. Results: Results of the meta-analysis demonstrate a strong correlation between LE and BICR estimates of treatment effect (R = 0.947). Further, differences between treatment groups in discordance rates predict the differences between LE and BICR estimates of treatment effect supporting the use of a sample-based BICR on a subgroup of patients. Conclusion: The meta-analysis demonstrates that local evaluation (LE) provides a reliable estimate of the treatment effect with little evidence for systematic evaluation bias. Therefore, when a trial is blinded or a large effect on PFS is observed a BICR may not be warranted. When a BICR is warranted, a sample-based BICR may provide a reduction in operational complexity without compromising the credibility of trial results. While for large trials that are not adequately blinded a sample-based BICR may be recommended. A full BICR should be considered in the case of smaller trials or in situations in which there is a particular need to increase the confidence in the LE results. © 2011 Elsevier Ltd. All rights reserved.
White R.E.,XenoBiotic Laboratories Inc. |
Evans D.C.,Janssen Research and Development LLC |
Hop C.E.C.A.,Genentech |
Moore D.J.,Hoffman LaRoche |
And 3 more authors.
Xenobiotica | Year: 2013
The necessity of conducting traditional radiolabeled absorption, distribution, metabolism and excretion (ADME) studies in animals during development of new investigative agents has been questioned in a recent review. We present a compilation of the benefits of such studies in the understanding of the in vivo pharmacological activity and disposition of new drug candidates, including interpretation of preclinical toxicology findings, characterization of circulating metabolites, and determination of principal pathways of clearance. This understanding is valuable in anticipating the human disposition of the drugs and the planning of the clinical development program. Because of new technologies, evolving regulatory expectations, and increased scientific understanding of the disposition of drugs, the traditional design and timing of both animal and human ADME studies should be reviewed. Innovative "fit-for-purpose" studies may well be a better choice in a particular drug development program than a standard animal ADME "package". However, we submit that, at this time, radiolabeled animal ADME studies still provide a definitive and irreplaceable component of our understanding of the in vivo actions and behaviors of drugs and should continue to be performed prior to the exposure of large numbers of human subjects to investigative drugs. © 2013 Informa UK, Ltd.
PubMed | Hoffman LaRoche, University of Washington, Fred Hutchinson Cancer Research Center, Moffitt Cancer Center and 9 more.
Type: | Journal: Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation | Year: 2016
Daclizumab, a humanized monoclonal antibody, binds CD25 and blocks formation of the IL-2 receptor on T cells. A study of daclizumab as acute graft-versus-host disease (GVHD) prophylaxis after unrelated bone marrow transplantation was conducted before the importance of CD25
News Article | December 17, 2016
Receive press releases from Strathmore Who's Who: By Email Strathmores Whos Who Honors John T. Spitznagel as a 2016 Professional of the Year John T. Spitznagel has recently been recognized as a 2016 Professional of the Year by Strathmores Whos Who for his outstanding contributions and achievements in the field of Healthcare Sales and Marketing. New York, NY, December 17, 2016 --( John T. Spitznagel is Chief Executive Officer of NovaSom Inc., a leader in Obstructive Sleep Apnea (OSA) home testing located in Glen Burnie, Maryland. Mr. Spitznagel received a B.A. in English from Rider University and an M.B.A. in Marketing from Fairleigh Dickinson University. Mr. Spitznagel has over 45 years of experience as a healthcare executive, with extensive knowledge of sales, marketing, management and business development. Mr. Spitznagel was Chair and CEO of ESP Pharmaceutical, a start-up hospital focused pharmaceutical company. He was Chair and CEO of Esprit Pharmaceutical, a start-up urology focused pharmaceutical company. He was also CEO of Roberts Pharmaceutical, President of Reed & Cormick Pharmaceuticals,a division of Block Drug Co., CEO of the drug discovery company Biocryst Pharmaceutical, Senior Vice President of Marketing and Sales for Wyeth Labs and Marketing Director of Roche Labs, a division of Hoffman LaRoche. Mr. Spitznagel was the recipient of the Entrepreneur of the Year Award in 2005 and 2010 and the Winner of Director Marketing Association Best Direct Mail Campaign. In his leisure time, Mr. Spitznagel enjoys swimming, fishing and sailing. About Strathmore’s Who’s Who Strathmore's Who's Who publishes an annual two thousand page hard cover biographical registry, honoring successful individuals in the fields of Business, the Arts and Sciences, Law, Engineering and Government. Based on one's position and lifetime of accomplishments, we honor professional men and women in all academic areas and professions. Inclusion is limited to individuals who have demonstrated leadership and achievement in their occupation, industry or profession. New York, NY, December 17, 2016 --( PR.com )-- About John T. SpitznagelJohn T. Spitznagel is Chief Executive Officer of NovaSom Inc., a leader in Obstructive Sleep Apnea (OSA) home testing located in Glen Burnie, Maryland. Mr. Spitznagel received a B.A. in English from Rider University and an M.B.A. in Marketing from Fairleigh Dickinson University. Mr. Spitznagel has over 45 years of experience as a healthcare executive, with extensive knowledge of sales, marketing, management and business development. Mr. Spitznagel was Chair and CEO of ESP Pharmaceutical, a start-up hospital focused pharmaceutical company. He was Chair and CEO of Esprit Pharmaceutical, a start-up urology focused pharmaceutical company. He was also CEO of Roberts Pharmaceutical, President of Reed & Cormick Pharmaceuticals,a division of Block Drug Co., CEO of the drug discovery company Biocryst Pharmaceutical, Senior Vice President of Marketing and Sales for Wyeth Labs and Marketing Director of Roche Labs, a division of Hoffman LaRoche. Mr. Spitznagel was the recipient of the Entrepreneur of the Year Award in 2005 and 2010 and the Winner of Director Marketing Association Best Direct Mail Campaign. In his leisure time, Mr. Spitznagel enjoys swimming, fishing and sailing. www.novasom.com About Strathmore’s Who’s WhoStrathmore's Who's Who publishes an annual two thousand page hard cover biographical registry, honoring successful individuals in the fields of Business, the Arts and Sciences, Law, Engineering and Government. Based on one's position and lifetime of accomplishments, we honor professional men and women in all academic areas and professions. Inclusion is limited to individuals who have demonstrated leadership and achievement in their occupation, industry or profession. Click here to view the list of recent Press Releases from Strathmore Who's Who
News Article | November 4, 2016
Honor Roll Member, John T. Spitznagel Has Been Recognized by Americas Registry of Outstanding Professionals John T. Spitznagel of Glen Burnie, Maryland is an Honor Roll Member of Americas Registry of Outstanding Professionals. He has now been selected as the Professional of the Year 2016 for his accomplishments in the field of Healthcare Sales and Marketing. Farmingdale, NY, November 04, 2016 --( The Professional of the Year membership in America’s Registry is awarded to those candidates who have achieved recognition in their respective industry or profession for their accomplishments and who have established a commendable reputation. This membership honors individuals who have reached the highest standards of business practices and, as such, should be recognized for their standings in the business world. America’s Registry is pleased to honor John T. Spitznagel for his accomplishments in the field of Healthcare Sales and Marketing. Farmingdale, NY, November 04, 2016 --( PR.com )-- John T. Spitznagel was born in Auburn, New York. He earned his B.S. in English at Rider University and his M.B.A. in Marketing at Fairleigh Dickinson University. Currently, he is the Chief Executive Officer of NovaSom Inc., www.novasom.com . Mr. Spitznagel’s illustrious career began as Chair and CEO of Esprit Pharmaceutical, then Chair and CEO of ESP Pharmaceutical. He continued on to become CEO of Roberts Pharmaceutical, President of Reed & Carnrick Pharmaceuticals, a division of Block Drug Company, CEO of Biocryst Pharmaceutical Company and then Senior Vice President of Marketing and Sales at Wyeth Labs. Mr. Spitznagel later became Marketing Director of Roche Labs, a division of Hoffman LaRoche, Product Manager and Sales Representative of Warner Chilcolt Labs, a division of Warner Lambert. Mr. Spitznagel’s honors and awards include Entrepreneur of the Year 2005 and 2010, Top 50 Best Companies to Work For, 2004, ESP and Winner of the Direct Marketing Association Best Direct Mail Campaign. His leisure time is spent swimming, fishing and sailing.The Professional of the Year membership in America’s Registry is awarded to those candidates who have achieved recognition in their respective industry or profession for their accomplishments and who have established a commendable reputation. This membership honors individuals who have reached the highest standards of business practices and, as such, should be recognized for their standings in the business world. America’s Registry is pleased to honor John T. Spitznagel for his accomplishments in the field of Healthcare Sales and Marketing.
Fielden M.R.,Hoffmann-La Roche |
Fielden M.R.,Amgen |
Hassani M.,Hoffmann-La Roche |
Hassani M.,Roger Williams Medical Center |
And 9 more authors.
Experimental and Toxicologic Pathology | Year: 2010
A number of drugs and drug candidates, including fenfluramine and ergot derivatives, are associated with valvulopathy in humans; however, these responses are poorly predicted from animal studies. In vitro and in vivo evidence suggests that these compounds exert their pathological effect through activation of serotonin 2B receptor (5HT2BR) signaling. However, the variable effect of fenfluramine and other 5HT2BR agonists in rodents has cast doubt on the relevance of animal findings to predicting human risk. Herein, a candidate compound, RO3013, induced subendocardial cell proliferation in the mitral and tricuspid valves in rats after only 3 days of daily dosing. Additionally, there was a treatment-related increase in immunostaining of the proliferation marker Ki67, and phosphorylated Smad3 in the heart indicative of TGFΒ signaling co-localized with 5HT2BR expression. To substantiate the hypothesis that RO3013-induced valvular proliferation is secondary to 5HT2BR activation, the compound was evaluated in vitro and found to bind to the human 5HT2BR with a Ki of 3.8γM; however, it was virtually devoid of agonist activity in a functional assay in human cells. By contrast, RO3013 bound to the rat 5HT2BR with a Ki of 1.2γM and activated the receptor with an EC50 of 0.5γM. This agonist potency estimate is in good agreement with the free plasma concentrations of RO3013 at which valvular proliferation was observed. These results suggest that the rat may be susceptible to 5HT2BR-mediated valvular proliferation similar to humans; yet, the significant differences between binding and functional activities can be a possible explanation for the observed species-selective receptor responses. © 2009 Elsevier GmbH.