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Neurodegenerative diseases are a function of matrix breakdown:how to rebuild extracellular matrix and intracellular matrix

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Neural Regeneration Research 2018年第7期13卷 1185-1186页

作者： Greg Maguire bio Regenerative Sciences Inc.Neo GenesisInc.

Matrix within cells,the cytoskeleton,and that which surrounds cells,the extracellular matrix（ECM）,are connected to one another through a number of receptors inc.uding those in primary cilia,serving as an important chemical and physical signaling system：Mechanical forces generated through the matrix play a critical role in determining the form and function of tissues（Hughes et al.,2018）.

关键词： ECM Neurodegenerative diseases are a function of matrix breakdown:how to rebuild extracellular matrix and intracellular matrix

Systems biology approach to developing S^2RM-based “systems therapeutics” and naturally induced pluripotent stem cells

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World Journal of Stem Cells 2015年第4期7卷 745-756页

作者： Greg Maguire Peter Friedman SRM Living Foundry Bio Regenerative Sciences Inc.

The degree to, and the mechanisms through, which stem cells are able to build, maintain, and heal the body have only recently begun to be understood. Much of the stem cell's power resides in the release of a multitude of molecules, called stem cell released molecules(SRM). A fundamentally new type of therapeutic, namely "systems therapeutic", can be realized by reverse engineering the mechanisms of the SRM processes. Recent data demonstrates that the composition of the SRM is different for each type of stem cell, as well as for different states of each cell type. Although systemsbiology has been successfully used to analyze multiple pathways, the approach is often used to develop a small molecule interacting at only one pathway in the system. A new model is emerging in biology where systems biology is used to develop a new technology acting at multiple pathways called "systems therapeutics". A natural set of healing pathways in the human that uses SRM is instructive and of practical use in developing systems therapeutics. Endogenous SRM processes in the human body use a combination of SRM from two or more stem cell types, designated as S2 RM, doing so under various state dependent conditions for each cell type. Here we describe our approach in using statedependent SRM from two or more stem cell types, S2 RM technology, to develop a new class of therapeutics called "systems therapeutics." Given the ubiquitous and powerful nature of innate S2RM-based healing in the human body, this "systems therapeutic" approach using S2 RM technology will be important for the development of anti-cancer therapeutics, antimicrobials, wound care products and procedures, and a number of other therapeutics for many indications.

关键词： Stem cell Paracrine Growth factors Pluripotency Wound healing Cancer

Amyotrophic lateral sclerosis as a protein level,non-genomic disease:Therapy with S2RM exosome released molecules

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World Journal of Stem Cells 2017年第11期9卷 187-202页

作者： Greg Maguire bio Regenerative Sciences Inc.La JollaCA 92037United States

Amyotrophic lateral sclerosis(ALS) is a rapidly progressing neurodegenerative disease that leads to death. No effective treatments are currently available. Based on data from epidemiological, etiological, laboratory, and clinical studies, I offer a new way of thinking about ALS and its treatment. This paper describes a host of extrinsic factors, inc.uding the exposome, that disrupt the extracellular matrix and protein function such that a spreading, prionlike disease leads to neurodegeneration in the motor tracts. A treatment regimen is described using the stem cell released molecules from a number of types of adult stem cells to provide tissue dependent molecules that restore homeostasis, inc.uding proteostasis, in the ALS patient. Because stem cells themselves as a therapeutic are cumbersome and expensive, and when implanted in a host cause aging of the host tissue and often fail to engraft or remain viable, only the S2 RM molecules are used. Rebuilding of the extracellular matrix and repair of the dysfunctional proteins in the ALS patient ensues.

关键词： Amyotrophic lateral sclerosis Proteins Stem cells Extracellular matrix Exposome Heat shock protein Stem cell molecules

5-Hexylidene-4-propylamino-1,5-dihydroimidazol-2-one formed from Cu-catalyzed oxidation with implication for the structure of a His-Lys cross-link

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Chinese Chemical Letters 2007年第9期18卷 1025-1028页

作者： Yahu A.Liu Medicinal Chemistry ChemBridge Research Laboratories Inc.San DiegoCA 92127USA

Cu-catalyzed oxidation of 5-hexyl-1,3-dihydroimidazo-2-one (1) in the presence of propylamine, as surrogates for the oxidized His side chain and Lys side chain, was investigated. 5-Hexylidene-4-propylamino-1,5-dihydroimidazol-2-one (2), a model His-Lys cross-link product, was isolated and structurally characterized by NMR and mass spectrometry.

关键词： His-Lys cross-link Metal-catalyzed protein oxidation Oxidative stress

Development of a physiologically relevant dripping analytical method using simulated nasal mucus for nasal spray formulation analysis

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Journal of Pharmaceutical Analysis 2016年第5期6卷 283-291页

作者： Tina Masiuk Parul Kadakia Zhenyu Wang Respiratory Product Development Merck Research Laboratories Merck & Co.Inc.

Current methods for nasal spray formulations have been elementary evaluating the dripping characteristics of a formulation and have not assessed the behavior of the nasal formulation in the presence of varying types of mucus depending on the indication or diseased state. This research investigated the effects of nasal mucus on the dripping behavior of nasal formulations and focused on developing an improved in vitro analytical test method that is more physiologically relevant in characterizing nasal formulation dripping behavior. Method development was performed using simulated nasal mucus preparations for both healthy and diseased states as coatings for the dripping experiment representing a wide range of viscosity. Factors evaluated during development of this in vitro test method inc.uded amount of mucus, application of mucus, drying times, and compatibility of the mucus on a C18 Thin Layer Chromatography(TLC) substrate. The dripping behavior of nasal formulations containing a range of 1%Avicel to 3.5% Avicel was assessed by actuating the nasal spray on a perpendicular TLC plate coated with either healthy or diseased simulated nasal mucus. After actuation of the nasal spray, the dripping of the formulation on the coated TLC plate was measured after the plate was repositioned vertically. The method that was developed generated reproducible results on the dripping behavior of nasal formulations and provided critical information about the compatibility of the formulation with the nasal mucus for different diseased states, aiding in nasal spray formulation development and physical characterization of the nasal spray.

关键词： Nasal spray Nasal spray dripping method Nasal mucus Nasal formulation dripping

Nogo receptor expression in microglia/macrophages during experimental autoimmune encephalomyelitis progression

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Neural Regeneration Research 2018年第5期13卷 896-907页

作者： Amani A.Alrehaili Jae Young Lee Maha M.Bakhuraysah Min Joung Kim Pei-Mun Aui Kylie A.Magee Steven Petratos Department of Neuroscience Central Clinical School Monash University Department of Clinical laboratories College of Applied Medical Sciences Taif University Toolgen inc. Gasan Digital-Ro

Myelin-associated inhibitory factors within the central nervous system（CNS） are considered to be one of the main obstacles for axonal regeneration following disease or injury. The nogo receptor 1（NgR1） has been well documented to play a key role in limiting axonal regrowth in the injured and diseased mammalian CNS. However, the role of nogo receptor in immune cell activation during CNS inflammation is yet to be mechanistically elucidated. Microglia/macrophages are immune cells that are regarded as pathogenic contributors to inflammatory demyelinating lesions in multiple sclerosis（MS）. In this study, the animal model of MS, experimental autoimmune encephalomyelitis（EAE） was induced in ngr1^＋/＋ and ngr1^–/– female mice following injection with the myelin oligodendrocyte glycoprotein（MOG_（35–55）） peptide. A fatemap analysis of microglia/macrophages was performed throughout spinal cord sections of EAE-induced mice at clinical scores of 0, 1, 2 and 3, respectively（inc.easing locomotor disability） from both genotypes, using the CD11 b and Iba1 cell markers. Western immunoblotting using lysates from isolated spinal cord microglia/macrophages, along with immunohistochemistry and flow cytometric analysis, was performed to demonstrate the expression of nogo receptor and its two homologs during EAE progression. Myelin protein engulfment during EAE progression in ngr1^＋/＋ and ngr1^–/– mice was demonstrated by western immunblotting of lysates from isolated spinal cord microglia/macrophages, detecting levels of Nogo-A and MOG. The numbers of M1 and M2 microglia/macrophage phenotypes present in the spinal cords of EAE-induced ngr1^＋/＋ and ngr1^–/– mice, were assessed by flow cytometric analysis using CD38 and Erg-2 markers. A significant difference in microglia/macrophage numbers between ngr1^＋/＋ and ngr1^–/– mice was identified during the progression of the clinical symptoms of EAE, in the white versus gray matter regions of the spinal cord. This difference was unrelated to the e

关键词： microglia experimental autoimmune encephalomyelitis nogo receptor myelin-associated inhibitory factors (MAIFs) Nogo A neural regeneration

Simultaneous determination of kolliphor HS15 and miglyol 812 in microemulsion formulation by ultra-high performance liquid chromatography coupled with nano quantity analyte detector

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Journal of Pharmaceutical Analysis 2016年第1期6卷 11-17页

作者： Honggen Zhang ZhenyuWang Oscar Liu Merck Research laboratories Merck & Co. Inc. Rahway NJ 07065 USA

A novel method for simultaneous determination of kolliphor HS15 and miglyol 812 in microemulsion formulation was developed using ultra-high performance liquid chromatography coupled with a nano quantitation analytical detector （UHPLC-NQAD）. All components in kolliphor HS15 and miglyo1812 were well separated on an Acquity BEH C18 column. Mobile phase A was 0.1% trifluoroacetic acid （TFA） in water and mobile phase B was acetonitrile. A gradient elution sequence was programed initially with 60% organic solvent, slowly inc.eased to 100% within 8 min. The flow rate was 0.7 mL/min. Good linearity （r 〉 0.95） was obtained in the range of 27.6-1381.1 μg/mL for polyoxyl 15 hydroxystearate in kolliphor HS15, 0.8-202.0 μg/mL for caprylic acid triglyceride and 2.7-221.9μg/mL for capric acid triglyceride in miglyol 812. The relative standard deviations （RSD） ranged from 0.6% to 1.7% for intra-day precision and from 0.4% to 2.7% for inter-day precision. The overall recoveries （accuracy） were 99.7%-101.4% for polyoxyl 15 hydroxystearate in kolliphor HS15, 96.7%-99.6% for caprylic acid triglyceride, and 94.1%- 103.3% for capric acid triglyceride in miglyol 812. Quantification limits （QL） were determined as 27.6 μg/ mL for polyoxy115 hydroxystearate in kolliphor HS15, 0.8 μg/mL for caprylic acid triglyceride, and 2.7 μg/ mL for capric acid triglyceride in miglyol 812. No interferences were observed in the retention time ranges of kolliphor HSI5 and miglyol 812. The method was validated in terms of specificity, linearity, precision, accuracy, QL, and robustness. The proposed method has been applied to microemulsion for- mulation analyses with good recoveries （82.2%-103.4%）.

关键词： Kolliphor HS15 Miglyol 812 Ultra-high performance liquid chromato-graphy (UHPLC) Nano quantitation analytical detector(NQAD)

Development and validation of a GC–FID method for quantitative analysis of oleic acid and related fatty acids

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Journal of Pharmaceutical Analysis 2015年第4期5卷 223-230页

作者： Honggen Zhang Zhenyu Wang Oscar Liu Merck Research laboratories Merck & Co. Inc.

Oleic acid is a common pharmaceutical excipient that has been widely used in various dosage forms. Gas chromatography （GC） has often been used as the quantitation method for fatty acids normally requiring a derivatization step. The aim of this study was to develop a simple, robust, and derivatization-free GC method that is suitable for routine analysis of all the major components in oleic acid USP-NF （United States Pharmacopeia-National Formulary） material. A gas chromatography-flame ionization detection （GC-FID） method was developed for direct quantitative analysis of oleic acid and related fatty acids in oleic acid USP-NF material. Fifteen fatty acids were separated using a DB-FFAP （nitroterephthalic acid modified polyethylene glycol） capillary GC column （30 m × 0.32 mm i.d.） with a total run time of 20 rain. The method was validated in terms of specificity, linearity, precision, accuracy, sensitivity, and robust- ness. The method can be routinely used for the purpose of oleic acid USP-NF material analysis.

关键词： Oleic acidFatty acidsGas chromatographyMethod developmentDerivatization-ffee

Growth performance,gastrointestinal weight,microbial metabolites and apparent retention of components in broiler chickens fed up to 11%rice bran in a corn-soybean meal diet without or with a multi-enzyme supplement

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Animal Nutrition 2019年第1期5卷 41-48页

作者： Juan Sanchez Aizwarya Thanabalan Tanka Khanal Rob Patterson Bogdan A.Slominski Elijah Kiarie Department of Animal biosciences University of Guelph Canadian bio-Systems inc. Department of Animal Science University of Manitoba

We investigated the effects of adding up to 11% rice bran(RB) in corn-soybean meal diets fed to broiler chickens without or with a multi-enzyme supplement(MES). The MES supplied xylanase, b-glucanase invertase, protease, cellulase, a-amylase and mannanase with targeted activity of 2,500, 300, 700, 10,0001,200, 24,000, and 20 U/kg of feed, respectively. The study used a two-phase feeding program(starter d 0 to 24; finisher, d 25 to 35) with RB added at 5% and 11%, respectively creating 4 diets in each phase Diets were iso-caloric and iso-nitrogenous and contained phytase(500 FTU/kg) and TiO_2 as a digestibility marker. Three hundred and sixty d-old male Ross 708 broiler chicks were placed in cages based on BW(15 birds/cage) and allocated to 4 diets(n = 6). Birds had free access to feed and water. Body weight and feed intake were recorded. Excreta samples were collected 3 d prior to the end of each phase for apparent retention(AR) of components. Samples of birds were sacrificed on d 24 and 35 for gut weight and ceca digesta for organic acid content. There was no interaction(P > 0.10) between RB and MES on BWG and FCR in starter or finisher phase. In finisher phase, birds fed MES had better BWG(961 versus 858 g) and FCR(1.69 versus 1.86) than birds fed non-MES diets(P inc.eased gizzard weight and reduced final BW whereas MES improved growth and energy utilization.

关键词： Broiler chickens Growth performance Gut weight Microbial activity Multi-enzyme supplement Rice bran