text:
The work of the association relies on the many contributions of members such as you. Volunteer leaders are the mainstay of any professional organization.
The Academy's Nutrition Care Manual is a comprehensive online resource that covers all aspects of nutrition management. NCM's user-friendly resources include:
Learn more »
Special Feature
The Nutrition Care Process in Pediatric Practice
This easy-to use guide shows how to incorporate standardized language—the International Dietetics & Nutrition Terminology (IDNT)—into everyday pediatric practice.
« Back To Eat Right Weekly - April 6
The National Fruit and Vegetable Alliance, of which ADA is a member, reported recently on its major undertaking, "Let's Move Salad Bars 2 Schools." This project helps extend efforts of ADA's School Nutrition Services DPG, providing grants to schools to give students access to healthy fruit and vegetables.
The NFVA reported on "Let's Move Salad Bars 2 Schools" at the 20th annual meeting of the Produce for Better Health Foundation. Headed by ADA member Elizabeth E. Pivonka, RD, the PBH is a partnership of growers and processors. Known for the successful More Matters programs, PBH is committed to finding ways for Americans to enjoy fruits and vegetables.
Copyright © 1995-2014 | Academy of Nutrition and Dietetics, All Rights Reserved. Privacy Statement | Terms and Conditions | Editorial Policy | Advertising & Sponsorship | Careers | Contact Us


text:
Skip to content
In association with
By MCN -
New bikes
27 April 2012 09:23
The new Suzuki GSX-R1000 is one of only two current superbikes (the other the Honda Fireblade) that doesn't have traction control.
For most of us mere mortals, that means having to be a little more careful on the throttle than on an electronics-laiden Aprilia RSV4, or any of the other 1000cc superbikes.
For 500GP winner Simon Crafar it means he can perform full-lock powerslides while exiting a second gear 80mph corner at Almeria.
MCN's Adam Child road the bike at the launch at the Miami Speedway track: ""Though peak power and torque is unchanged, the bulge of midrange has been moved lower down the rev-range which makes riding it fast very, very easy.
"Coming out of slow corners - corners that would mean first gear on an S1000RR or ZX-10R – you just leave it in second and drive.
"The brakes on the old bike were a weak point on track but the new ones are really strong. I've had no fade, and there are some massive braking areas here.
Upload stories, photos or videos direct to the site, or email email@example.com.
Or call us on 01733 468842 (+44 1733 468842)
Add your comment – Video: Simon Crafar powerslides the new Suzuki GSX-R1000
Sign in You must be signed in to submit a comment.
I can do that! On my Playstation that is.
27 January 2013 18:10
Cool video and I kind of get where they're coming from. Instead of TC I'd like to see a 2WD Superbike. I remember a few years back someone put a hydraulic drive system onto the front of an R1, they reckoned it could lap a fair bit quicker than a standard RWD R1 in the rain. That would help with traction and wheelies. Seems like that might be a better way to go than building a bike with 200bhp then using electronics to slow it down again.
03 May 2012 16:54
seriously skilful stuff
don't try this at home kids !!
27 April 2012 19:19
Gee Ma are those real slides?
27 April 2012 15:59
27 April 2012 14:30
Save money by comparing quotes. It's quick and easy
With over 18,000 products, MCN shop is the best place to buy your biking kit
Motorcycle Cover | Cheap Motorcycle Insurance | Moped Insurance | Scooter Insurance | 125 Insurance | Work Experience | Complaints Policy
Car Magazine | Empire | FHM | Parkers | LRO | Today's Golfer | Zoo
Bauer Consumer Media Ltd (company number 01176085, 1 Lincoln Court, Lincoln Road, Peterborough, England PE1 2RF)

subset: commoncrawl


text:
Better Than: That time where I could've seen Ryan Adams open for Oasis in 2008 and passed on it like "Eh, they'll both be back around soon enough" and then waited six years for a chance to see Adams for the first time. Ryan Adams is a charmer. That's something that got lost in the shuffle back whe ... More >>
Fact: Any band who uses Times New Roman on their album cover isn't going to make it in the music industry. (Feel free to prove us wrong, though). Smart typography selection is as important to a band's image as having a memorable logo. In 1937, industrial designer and typographer Gerry Powell, desig ... More >>
Your copy of Purple Rain sounds terrible. That's no knock against the music, that urgent neon bug-funk zeitgeist-bomb Prince straddled and launched and screamed on as it blew up the world, kind of like what Slim Pickens' did to that nuke in Dr. Strangelove but with more cream. That music still sti ... More >>
Cruise through this index of the Voice's top ten albums from 1971 through 2013, according to its annual Pazz & Jop critics' poll, started by Robert Christgau. The poll has run every year since '71, except for a three-year hiatus after that first year. The prominence of legacy artists on the inaugu ... More >>
Today and Saturday, Jay-Z and Justin Timberlake take the stage at Yankee Stadium for their Legends of Summer tour. While they're both certainly living legends in their own right, it's interesting to look back on how truly different both Hov and Timberlake's summers have been over the years, and yet ... More >>
Kings of the Stone Age
Gagosian Gallery hosts a display of the late artist's spot paintings
The Black Angels take time off from planning their festival to play ours
Bekah CopeJamin and Jake With their savage follicles and beer-soaked, bruised take on punk, Nashville's JEFF The Brotherhood seem like the type of boys that would steal both your whiskey and your sister's virtue, and still earn a standing invitation to play your basement. But this might be an unfai ... More >>
Yes In My Backyard is a semiweekly column showcasing MP3s from new and emerging local talent. O.C. and A.G. are New York hip-hop legends holding their own. The two members of longstanding pan-borough collective Diggin' In The Crates (the same one that spawned Fat Joe and Big L) still sound d ... More >>
Mountain Dew's MP3-as-single imprint Green Label Sound has an exclusive download of the Amazing Baby single "Supreme Being." The psych-pop track was a shimmering stand-out from the Brooklyn band's 2008 four-song EP Infinite Fucking Cross, so it was surprising when the moony floater didn't make th ... More >>
--Noel Gallagher has posted a statement regarding his reasons for quitting Oasis minutes before the band was to go onstage at the Rock en Seine festival in France. On the band's website, Gallagher wrote that "the level of verbal and violent intimidation towards me, my family, friends and comr ... More >>
all photos by Rebecca Smeyne While you were busy shrieking at the East River stormclouds this past weekend, some of our locals were overseas for Fuji Rock, a gorgeously run three-day Japanese music festival. There were approximately 130,000 attendees, Oasis, Weezer, and Franz Ferdinand as headliner ... More >>
Catching up with our finest purveyor of impromptu noise-punk trilogies
Indeed. Earlier this week came the terrible news of Adam Yauch's cancer diagnosis and the subsequent cancellation of all upcoming Beastie Boys tour dates -- including their opening-night headlining spot at next week's All Points West festival in Jersey City. Their replacement: Jay-Z, making "his U.S ... More >>
Weezer Wednesday September 24 Madison Square Garden Photos by David Atlas There's a 1988 evaluation sheet from Rivers Cuomo's pre-Weezer metal band on Weezed.com. Not sure who wrote it, because there are three kinds of handwriting on tips like "gotta have a core following," and "need something s ... More >>
Everything greatand everything you forgot was lousyabout British guitar-rock
Vivid art-rockers Apollo Heights craft 'soul songs for people who hate our music'
Dancefloor chic with England's latest and greatest
Brit lads get things like naked girlfriends out in the open
With the Brit avant-garde, rust-belt saxophone, Braxton standards, Dylan standards
Musicians settle with Knitting Factory after protest over backstock, royalties, masters
Add to the many things not to be thankful for this year 13 CDs that will make your gorge rise
Sun City Girls tune their static into freaky Asian frequencies, keep changing the channel
Peppy Songs With Hooks and Beatsthe New Underground Craze!
Blues You Can Use Before History Uses You
DJ Music Builds Its Way Out of the Velvet-Rope Underground


text:
- freely available
- re-usable
Marine Drugs 2010, 8(9), 2493-2516; doi:10.3390/md8092493
Published: 15 September 2010
Abstract: N-Acetylglucosamine (GlcNAc) is a monosaccharide that usually polymerizes linearly through (1,4)-β-linkages. GlcNAc is the monomeric unit of the polymer chitin, the second most abundant carbohydrate after cellulose. In addition to serving as a component of this homogeneous polysaccharide, GlcNAc is also a basic component of hyaluronic acid and keratin sulfate on the cell surface. In this review, we discuss the industrial production of GlcNAc, using chitin as a substrate, by chemical, enzymatic and biotransformation methods. Also, newly developed methods to obtain GlcNAc using glucose as a substrate in genetically modified microorganisms are introduced. Moreover, GlcNAc has generated interest not only as an underutilized resource but also as a new functional material with high potential in various fields. Here we also take a closer look at the current applications of GlcNAc, and several new and cutting edge approaches in this fascinating area are thoroughly discussed.
1. Introduction
Carbohydrates, with the general formula Cm(H2O)n, are the most abundant organic compounds in living organisms. Also known as saccharides, carbohydrates are involved in a variety of biological events. Fox example, they serve as the main source and storage form of energy as well as structural components. Monosaccharides, the monomers of carbohydrates, serve as the main source of energy for metabolism (e.g., glucose) and are used in biosynthesis [1]. Their derivatives are also important components of coenzymes and the backbone of genetic molecules. Furthermore, carbohydrates are essential in cell interactions [2], signal transduction [3,4], antibody recognition [5], tumor metastasis [6], hormones [7] and musculoskeletal physiology [8,9].
N-Acetylglucosamine (GlcNAc), 2-acetamino-2-deoxy-β-d-glucose or 2-(acetylamino)-2-deoxy-d-glucose, is a monosaccharide derivative of glucose and is widely distributed worldwide. The molecular formula of this amino monosaccharide is C8H15NO6, and its molecular weight is 221.21. In general, it is a white and slightly sweet powder that melts at 221 °C. The solubility of GlcNAc is 25% in water, and 1% aqueous solutions are colorless and clear.
GlcNAc polymerizes linearly with (1,4)-β-linkages and is the monomeric unit of the polymer chitin, the second most abundant carbohydrate after cellulose. Chitin is present in arachnids, most fungal cell walls, insect exoskeletons, the shells of crustaceans and parts of invertebrates [10,11]. It is also present as an extracellular polymer of some microbes [12,13]. Isolated chitin exists as two allomorphs, namely α-chitin and β-chitin. α-chitin is the most abundant by far and is present in shrimp and crab shells. The rarer β-chitin is found in squid pens and is commercially more expensive. To date, it is not possible to obtain β-chitin either from solution or by in vitro biosynthesis.
In addition to being a structural component of homogeneous polysaccharides like chitin, GlcNAc is also a constituent of heterogeneous polysaccharides, such as murein [10–14] and hyaluronic acid (also called hyaluronan or hyaluronate, HA) [14]. Murein is the basic component of the bacterial cell wall and consists of crosslinked peptide chains with repeating GlcNAc and muramic acid residues [14]. HA is a linear heteropolysaccharide that is composed of repeating d-glucuronic acid and GlcNAc residues [14]. As early as 1943, HA was discovered in bovine eyes, and its defect causes disease. In fact, it is the major component of the extracellular matrix and is extensively distributed in connective, epithelial and neural tissues [15–20]. HA performs numerous roles in cell motility, inflammation and cancer metastasis [21]. In addition to its contributions in eukaryotes, HA is also a component of cell coatings in some bacteria [22]. HA can be produced by biochemical engineering techniques [23,24].
Chondroitin is a glycosaminoglycan that is composed of the same repeats as HA but is sulfated at position C-4 or C-6 of the N-acetylgalactosamine residues. It is also plentiful in connective tissues, particularly in blood vessels, bone and cartilage. In fact, GlcNAc and sulfated GlcNAc can be isolated from heparin or keratin sulfate [25,26]. Heparin, which is produced by basophils and mast cells, consists of a variably-sulfated repeating disaccharide unit and acts as an anticoagulant. The major repeating disaccharide unit of keratin sulfate, also called keratosulfate, is composed of galactose and sulfated GlcNAc. Keratin sulfate is distributed in the cornea, cartilage and bone and usually acts as a cushion in joints to absorb mechanical shock.
Moreover, GlcNAc molecules with various functional group substitutions serve as ingredients of cell surface decoration and/or are directly involved in cell interactions [5]. Indeed, these carbohydrates may increase the specificity and interaction force between cells. For example, blood type usually depends on the oligosaccharides present of the surface of erythrocytes in human beings, and GlcNAc appears to be the basic element of the H antigen in ABO blood grouping [5]. GlcNAc was also identified in the two epitopes Sialy-Lewis X and Sialy-Lewis A. They are ligands that capture selectins for adhesion, but they also contribute significantly to metastasis [6,27–29].
In addition, glycoproteins that contain GlcNAc can be found in the mucous membranes of the digestive tract. However, saccharide modification may promote pathogen binding. For example, GlcNAc-β-(1–4)-GlcNAc is present in membrane proteins of brain microvascular endothelial cells (BMECs) and acts as the ligand for Escherichia coli K1 outer membrane protein A (OmpA) to mediate bacterial transport through the blood-brain barrier in neonates [30,31].
Generally, oligosaccharide modification is proposed to play a role in structural stability and interaction with other molecules. In particular, O-GlcNAcylation affects performance, including activity, stability and/or localization [32]. O-linked GlcNAc usually modifies nuclear pore proteins that are involved in nuclear pore assembly and function [33]. However, N-GlcNAcylated glycoproteins are cleared more rapidly by lysosomal glycosidases [34].
In plants, GlcNAc has been found in bromelain, ricin agglutinin and abrus agglutinin [7,35–37]. In humans, GlcNAc is frequently observed in glycoproteins, such as tissue plasminogen activator [38]. It is also detected in mammalian growth factors and hormones [35], including follicle-stimulating hormone (FSH), luteinizing hormone (LH), human menopausal gonadotropin (hMP), pregnant mare serum gonadotropin, thyroid-stimulation hormone (TSH) and human chorionic gonadotropin (hCG).
Recently, GlcNAc and its derivates been utilized in dietary supplements and for therapeutic development [8,39–41] due to its unique characteristics. Toxicity tests reveal that GlcNAc is non-toxic, supporting the essential safety issue, and the half-life of GlcNAc with 20 g intravenous injection is 220 min [42]. While 800 mg administrated per os, the mean maximum concentration (Cmax) of GlcNAc in plasma was 162.7 ± 125.2 ng/mL and the time for maximum concentration (Tmax) was 1.56 ± 1.23 h [43]. It was further demonstrated that 54% of the administered glucosamine (GlcN) was excreted into the urine in one day, indicating the clearance pathway of GlcNAc [42]. Therefore, GlcNAc can potentially be utilized in many significant applications.
2. Production of N-Acetyl-d-glucosamine
In mammals, GlcNAc and GlcN are components of glycoproteins, proteoglycans, glycosaminoglycans (GAGs) and other connective tissue building blocks [15–20]. Despite being a building block of biomacromolecules, GlcNAc seldom exists in free form, except in human milk (at 600–1500 mg/mL) [44,45]. Because GlcNAc not only play a role in plant organogenesis and invertebrate embryogenesis [46] but also has therapeutic potential in the treatment of a wide variety of diseases, economic feed-stocks and high efficiency processes for producing GlcNAc are urgently needed in industry. Every year, about 100 billion tons of chitin are produced in nature [47–49], making chitin a suitable biomass resource for the production of GlcNAc. Using chitin as feed-stock, GlcNAc can be prepared through a process that is based on chitin hydrolysis. Figure 1 summarizes the processes of GlcNAc production to date.
2.1. Chemical methods to produce GlcNAc
GlcNAc has historically been commercially prepared by several companies via a process based on the acid hydrolysis of crude chitin. Generally, the chemical degradation of chitin can be performed through hydrolysis using a strong acid, such as HCl. The processing temperature and concentration of the acid should be carefully selected. These parameters must be high enough to sufficiently degrade chitin but not so high that the GlcNAc product is destroyed. Suitable reaction conditions include 15–36% HCl and about 40–80 °C. In this procedure, up to 6.42 g/L GlcNAc can be produced in 1 h [50]. However, there appears to be several problems in producing GlcNAc by the direct acid hydrolysis of chitin, including high cost, low yield (below 65%) and acidic waste created by the use of HCl [51].
An alternative process involves the use of GlcN. Chitin can be dissolved in concentrated HCl and heated in boiling water for 3 h. In such drastic conditions, the acetyl group of GlcNAc is removed. An additional N-acetylation reaction must then be performed to produce GlcNAc. In a typical preparation, GlcN is dissolved in 10% methanol and is N-acetylated with acetic anhydride in the presence of Dowex 1 (carbonate form). After filtration and crystallization, this process can generate an overall yield of 43% GlcNAc [52,53]. Another improved N-acetylation procedure was described by Zhan (2007). GlcN is N-acetylated using pyridine as a solvent in the presence of tributylamine and acetic anhydride. After a series of purification procedures, ~99% pure GlcNAc can be obtained. The yield of this method has been reported to be higher than 70% [54].
Although the production of GlcNAc by chemical methods is estimated to be sufficiently economic, the product is not considered a natural material due to its chemical modification [51]. In addition to N-acetylated products, trace amounts of O-acetylated and di-acetylated products have also been detected by LC-MS. Furthermore, solvent and tributylamine are present in the product, as determined by GC-MS (unpublished data). Thus, chemically produced GlcNAc usually tastes bitter due to these residual substances.
Several efforts have been made to improve the purity of GlcNAc produced by chemical methods. Ryosuke et al. (2000) combined the acid hydrolysis of chitin and ion-exchange membrane electrophoresis to prepare natural GlcNAc [55]. Zhan (2007) established a novel purification and recrystallization procedure to prepare GlcNAc with purity that was higher than 99.95% [54]. In addition, the ozone treatment has been developed [56]. Thus, it will be possible to use chemically modified GlcNAc as an inexpensive resource in food additives, cosmetics and pharmaceuticals in the future.
2.2. Enzymatic methods to produce GlcNAc
GlcNAc produced by chemical methods is not widely commercialized, not only due to technical reasons but also because of environment concerns. The large quantities of chemical waste resulting from chemical processes are not environmentally friendly. However, the enzymatic hydrolysis of chitin can produce GlcNAc under mild conditions.
The enzymes degrading chitin, chitinases, distributes in various organisms [10]. They involves in the physiological and pathological functions [57–62]. The ensemble of chitinolytic enzymes can contain endochitinases (EC 188.8.131.52), exochitinases (EC 22.214.171.124), chitobiosidases (EC 126.96.36.199) and N-acetylglucosaminidases (NAGases) (EC 184.108.40.206) that are produced and secreted from either a prokaryotic or eukaryotic organism [10]. Usually, the endochitinases within microbes cleave chitin randomly at internal sites on the glycosidic bonds between GlcNAc residues, and such generates the soluble and low molecular weight oligosaccharides (e.g., tetramers, trimers and dimers). To increase the solubilization of chitin is essential as the first step. Then, the exochitinases or chitobiosidases in some organisms catalyze the progressive release of dimers, starting at the non-reducing end of chitin. Finally, NAGases cleave the oligomers and dimers, which are produced by endochitinases and exochitinases, to generate GlcNAc [63,64]. Several crude enzymes isolated from Trichoderma viride, Aspergillus niger, Carcica papaya L. and Aeromonium have been found to degrade β-chitin and to produce GlcNAc efficiently [51]. In contrast, some other microbes, such as Trichoderma hamatum AB10282 strain (FERM BP-10623) or Trichoderma harzianum AB10283 strain (FERM BP-10624), produce GlcNAc in the medium without the presence of chitin or chitin derivatives [65].
For the industrial production of GlcNAc, a large quantity of enzymes can be purified from the mass production of microbes or genetically engineered microorganisms [66–68]. However, owing to the low physiological content of chitinase in organisms, the direct purification of chitinase, even from the fermentation broth of genetically engineered microbes, is expensive. Some commercial, unspecific, crude enzymes, such as cellulose, lysozyme, papain and lipase, also degrade chitin due to the presence of endo- and exochitinase in crude enzyme preparations [69–71]. These unusual enzymes are inexpensive, and large amounts of them can be easily obtained commercially.
Several enzymatic processes can be performed to produce GlcNAc. Sashiwa et al. (2002) described the effective production of GlcNAc from flake type α-chitin by crude enzymes derived from Aeromonas hydrophila H2330 [72]. In this study, the selective production of GlcNAc from α-chitin was achieved with a yield up to 77% in 10 days. Although high yield and purity was achieved, low productivity was the crucial defect of this method. Another process has been described by Pichyangkura et al. (2002) in which crude chitinases from Bacillus licheniformis SK-1 were used to digest α-chitin powder for the production of GlcNAc. However, only a 41% yield of GlcNAc was achieved along with the production of chitobiose [73].
Poor yields were reported in these studies using α-chitin as a substrate. The major impediment of the enzymatic hydrolysis process is the extremely low susceptibility of natural α-chitin due to its high crystallinity. In another experiment, the hydrolysis of β-chitin, which shows good swelling properties compared with α-chitin, was accomplished by crude enzymes (e.g., cellulose T and A), and a yield of nearly 100% was achieved [74]. It is worth noting that the amorphous portion of chitin is easily hydrolyzed, and the remaining tightly packed chitin is hydrolyzed slowly.
Almost 100% pure GlcNAc was produced using crude enzymes from A. hydrophila H2330 [72], while chitobiose was synergistically produced using Bacillus chitinases [73]. This finding indicates that endochitinase, exochitinase and NAGase are necessary for the complete digestion of chitin, and higher NAGase activity results in a higher purity of GlcNAc. Thus, the chitin crystalline structure and the enzymatic composition of the reaction are two crucial factors in the production of GlcNAc through enzymatic methods.
2.3. Improved enzymatic methods to produce GlcNAc
The ability of chitin to act as a substrate for enzymatic hydrolysis might be improved by treatments that lead to its decrystallization. Several productive processes have been used to improve the purity and productivity of GlcNAc. Most of these processes focus on the modification of the chitin crystal structure because decrystallized chitins are more susceptible to enzymatic hydrolysis than natural chitin.
The crystallographic parameters of chitin reveal that there are two anti-parallel molecules per unit cell in α-chitin, while only one is present in β-chitin. The proposed crystal structures of α-chitin and β-chitin are organized into sheets that are tightly held by a number of intra-sheet hydrogen bonds. In addition, some inter-sheet hydrogen bonds are also present in α-chitin but not in β-chitin. Thus, α-chitin is thermodynamically more stable than β-chitin [75]. When β-chitin crystals are destroyed during acid swelling, new crystals of α-chitin are produced during recrystallization. The crystallinity of different chitin substances can be detected by X-ray diffractograms [76]. The limited enzymatic hydrolysis of natural chitin may be attributed to a limited accessibility of β-glycosidic bonds in the interior of the chitin crystal structure. Thus, β-chitin is degraded much more readily than α-chitin due to its weak intermolecular forces [77]. Sashiwa et al. (2001) obtained GlcNAc from 1% β-chitin by hydrolysis with various commercial enzymes, such as cellulose, hemicellulase, papain, lipase and pectinase. The highest yield (~76%) was obtained after eight days of hydrolysis with 20 mg/mL of cellulose [51].
Heat-treating chitin in a hydrophobic solvent or surfactant is suggested to weaken the hydrogen bonding or hydrophobic interactions of the crystal structure and, thus, facilitate the action of enzymes for decomposition. Chitin colloidized is better access for enzymes digestion [78]. Kawasaki et al. treated chitin using straight-chain hydrocarbons, surfactants (e.g., Triton X-100 or Tween 20) and urea to loosen the chitin structure and then heated or ultrasonicated the mixtures. These treatments increase the conversion ratio of chitin [79].
It is also possible to efficiently decompose chitin by exposing it to compounds that destroy its crystal structure. However, hydrogen bonding and hydrophobic interactions are generally necessary to maintain the three-dimensional structure of enzymes as well. Thus, chitin-degrading enzymes may be inactivated under these strict conditions. Enzyme reactions must be performed under conditions that their catalytic activities remain, such as with low concentrations of chemical agents. Recently, some novel sodium dodecyl sulfate (SDS)-resistant endochitinases and thermostable chitinases were found in the fermentation broth of bacteria [80–82]. These enzymes could be applied to improve the hydrolytic efficiency of chitin in the future.
The most common source of chitin for producing GlcNAc is shellfish biomass. However, the supply of shrimp and crab shells is highly dependent on seasonal and environmental factors, leading to unpredictable limitations on production capacity. Another option to stabilize the chitin source for the production of GlcNAc is to utilize a non-shellfish chitin source. In several typical fungal species, at least 15% of chitin exists in their cell walls. Chen et al. established a method to produce chitin by culturing an Actinomucor taiwanensis fungus. The highest yield of chitin was achieved at 1.5 g/L using their fermentation method [83]. The crystal structure of fungal chitin has not yet been characterized, although it is hypothesized to be highly susceptible to enzymatic degradation.
GlcNAc can be produced using fungal biomass as a substrate. Bohlman et al. (2004) obtained GlcNAc of high purity by degrading fungal chitin using either acid or enzymes. The concentration of GlcNAc obtained by acid degradation was 6.42 mg/mL, while that obtained by enzymatic treatment was 4.04 mg/mL [50,84]. Raetz et al. (1998) compared several chitins from different sources and tested their susceptibility to enzymes isolated from Penicillium janthinellum P9. Their results also confirm that fungal chitin has a higher susceptibility to hydrolysis than arthropod chitin [85].
A continuous production system used to obtain GlcNAc from chitin was described by Louise et al. This system involves five essential unit operations: (1) the fermentation unit, which is required for both cell culture and the production of chitinolytic enzymes; (2) the enzyme recovery unit; (3) the pretreatment of the chitin substrate; (4) the two-stage chitin-hydrolysis reactor; and (5) the final purification unit to obtain GlcNAc [86].
In the first unit, Serratia marcescens QM B1466 is used to produce chitinolytic enzyme ensembles. After induction for four to six days, the chitinase activity in the medium exceeds 105 U/mL, and the NAGase activity in the medium is two-fold higher than that of chitinase. In the second unit, chitinolytic enzyme ensembles are concentrated using colloidal chitin adsorption or ultrafiltration. In the third unit, mechanical methods, such as ball milling and hammer milling, are used to increase the surface area of chitin. Physical-chemical methods, such as autoclaving and steam exploding, are then used to swell the chitin particles, and treatment with dimethyl acetimide and weak cosolvents are explored to disrupt the chitin inter-chain hydrogen bonds.
In the fourth unit, a novel two-stage chitin-hydrolysis bioreactor was designed to maintain a high rate of chitin hydrolysis and GlcNAc production. The configuration of the bioreactor is illustrated in Figure 2. Enzyme ensembles are pumped into the first reactor, which contains pretreated chitin particles as a packed bed or fluidized bed. Chitinases are adsorbed onto chitin beads, and the beads are steadily hydrolyzed. NAGase, which is not adsorbed onto the chitin beads, and the synergetic soluble fractions are released into the second stirred reactor. The solid content is recycled back to the first reactor. The soluble fractions are completely hydrolyzed by NAGase, making GlcNAc the sugar product of the two-stage reactor. The enzyme catalyst stream is continuously recycled into the reactor after removing GlcNAc by cross-flow filtration.
In the fifth unit, a continuous cross-flow ultrafiltration cell is used to separate the GlcNAc product from the chitinolytic enzyme ensemble. GlcNAc, with a purity greater than 98%, is recovered in the filtrate, and no protein contaminants are detectable. However, the primary oligosaccharide contaminant in the product is chitobiose, as detected by HPLC. Further purification procedures are necessary to recover GlcNAc from the filtrate.
This continuous production system is capable of producing GlcNAc at an average productivity of 78 g/L/h. The two-stage bioreactors are operated continuously over a 10-day period, reaching steady-state production levels after 28 h, and this system is estimated to be an economic process for the production of GlcNAc.
2.4. Production of GlcNAc by biotransformation
Although economic processes to produce GlcNAc by enzymatic hydrolysis have been established, the necessity to arrange so many units is a limiting factor. Efforts to screen indigenous microorganisms from the soil for their ability to degrade chitin have been performed. Li et al. (2005) isolated a microbe, which was identified as Aeromonas caviae DYU-BT4, from a soil sample in Taiwan. In a fermentation test using chitin particles as a substrate, high chitinase activity (588.6 U/L) was detected, and the concentration of reducing sugar in the medium was 0.88 g/L. The major substance in the hydrolysate of chitin was GlcNAc. After optimization of the fermentation process, about 7.8 g/L of GlcNAc was produced using 2% of colloidal chitin as a substrate. This result indicates that it is possible to produce GlcNAc using whole microbes [87].
A novel chitin-degrading microbe named Chitinibacter tainanensis was isolated from a soil sample that was collected in southern Taiwan and was proven to produce GlcNAc [88]. The GlcNAc yield was 75% using α-chitin as a substrate and greater than 98% using β-chitin as a substrate. A needle-like crystal was produced after the broth was concentrated and crystallized, and the purity of the product was determined to be greater than 99%. It is suggested that some factors, called chitin-degrading factors (CDFs), can hydrolyze chitin and are released during the proliferation of C. tainanensis in the presence of chitin, with the concomitant production of acidic metabolite. Interestingly, the microbe is killed due to the acidic condition of the medium. However, CDFs still hydrolyze chitin, and the final product, GlcNAc, accumulates in the medium without being consumed by the microbe because it is dead [89].
Further studies indicate that CDFs are located on the surface of C. tainanensis. Thus, even if the microbes die, CDFs are still docked on the bacterial debris and maintain their chitinolytic activity. To determine how CDFs hydrolyze chitin, their endochitinase and NAGase activities have been measured. Both of these enzyme activities are found in live bacteria and in dead cell debris. The specific activity of NAGase is much higher than that of endochitinase. Therefore, GlcNAc is the only product of this biotransformation method. The mechanism of CDFs is similar to that of cellulosome from mesophilic bacteria [90] and is illustrated in Figure 3.
Another way to produce GlcNAc is through the genetic modification of microorganisms using glucose as a substrate. Metabolic pathways for GlcN and GlcNAc synthesis can be modified by genetic engineering techniques to increase enzyme activities, overexpress proteins, reduce product inhibition and increase the affinity of the substrate. A genetically engineered E. coli strain has been developed by Den et al. [91] in which GlcN-6-P acetyltransferase, GlcN-6-P synthase and GlcN-1-P acetyltransferase are overexpressed, while GlcN-6-P and GlcNAc-1-P uridyltransferase are suppressed. This microorganism contains additional genetic modifications that increase phosphoglucoisomerase, glutamine synthetase and glucose 6-P dehydrogenase activities and inactivate phosphofructokinase and genes encoding the enzymes responsible for glycogen synthesis to control the flux of metabolites. Because glucose is the major substrate for GlcNAc production, fructose is supplemented to maintain the energy supply for bacterial proliferation. In this process, a high concentration (exceeding 120 g/L) of GlcNAc was obtained after fermentation for 60 h and is, thus, the most efficient process by far.
3. Applications of GlcNAc
GlcNAc belongs to a large class of amino sugars that serve a number of functions and are located throughout the human system. For example, GlcNAc and its analogues, such as chitin and chitosan, are known as the carbon and/or nitrogen sources for creatures. In fact, they also could be used as a biomass. It has been shown that Rhodotorula glutinis is able to convert GlcNAc to biofuel [92,93]. The bioethanol could be yielded with the GlcNAc from chitin waste as a C6 carbon source [94].
GlcNAc is also a component of glycoproteins, proteoglycans, GAGs and other connective tissue building blocks [42]. GAGs and glycoproteins act as substrate materials for tissue repair and anti-inflammatory reactions [95]. When GlcNAc is present in the medium of pig gastric mucosal cells, it is incorporated into high molecular weight glycoproteins [96]. Additionally, after 168 h of oral administration of GlcNAc, about 25% of GlcNAc accumulates in tested animals [97]. This finding indicates that GlcNAc is an important component of biomacromolecular synthesis in the body.
Certain diseases appear to be related to abnormalities in the formation and utilization of amino sugars. Cell membranes, intercellular fluids and cell regeneration could all be affected by GlcNAc. In recent years, GlcNAc has been discovered to be a valuable pharmacological agent in the treatment of a wide variety of ailments.
3.1. Safety of GlcNAc
A large dose of GlcNAc (20 g) given intravenously to human volunteers results in neither toxicity nor alteration of blood glucose concentration [42,43]. A lack of insulin resistance was also found after oral administration of GlcNAc, even at a high dose. Systematic toxicological tests have been performed in animals, including the acute toxicity test, Ames test, micronucleus test of mouse bone marrow cells, abnormality test of mouse sperm, aberration test of mouse testis chromosomes, chronic lethal test, sub-chronic toxicity test (for 90 days) and traditional deformity-inducing test [98,99]. These experiments indicate that GlcNAc is a valuable compound with high safety in invaded, oral and topical usage. GlcNAc has been widely utilized as a nutritional supplement for therapeutic usage. It is classified as described in the following sections.
3.2. GlcNAc is used to treat joint damage
Articular cartilage is a matrix of proteoglycans, chondrocytes and collagens. It absorbs shock from mechanical forces and provides a smooth surface so that bone ends may glide easily across one another. Chondrocytes are known to contribute to the synovial fluid, which bathes the articular cartilage. Lack of proteoglycan precursors and synovial fluid leads to defects in the structure and function of skeletal joints and osteoarthritis occurs. The incidence increases with age and is found in most people over the age of 65 [100,101].
Many clinical trials have been performed to treat patients with joint disorders, including arthritic diseases, osteoarthritis, rheumatoid arthritis, cartilage damage, joint injury and degenerative joint disease (DJD). Preparations containing GlcNAc are delivered by parenteral, oral, transmucosal and topical administration. The results from these deliveries indicate that GlcNAc significantly enhances the prevention of joint damage [102–107]. Moreover, GlcNAc also inhibits elastase activity and superoxide release from human polymorphonuclear leukocytes [108,109].
3.3. GlcNAc is a potential candidate to treat inflammatory bowel disease (IBD)
Mucous membranes of the entire digestive tract are composed of epithelial cells, which have a high rate of turnover. The synthesis of glycoproteins and GAGs by the mucosa is considered to provide protection for the lining of bowel. IBD is a generic diagnosis that encompasses a number of bowel diseases, including ulceracolitis, chronic proctitis and Crohn's disease. IBD can involve the inflammatory disruption of vascular and matrix GAGs, and tumor necrosis factor-α appears to play an important role in inflammation [110–112]. The loss of GAGs from the intestinal wall results in fibrosis of the bowel. Tissue defects in the digestive tract result in food intolerance, in which food allergens are absorbed by the gastrointestinal tract, followed by the development of psoriasis [113,114].
GlcNAc is capable of enhancing the release of acid mucopolysaccharides by fibroblasts and restoring the formation of the protective structure of the gastrointestinal tract [115]. In addition, GlcNAc increases the elasticity of perivascular tissue, resulting in an increase in arterio-capillary blood flow. GlcNAc acts as a cytoprotective agent for restoring the integrity and normal function of the mucous membrane in humans [116,117]. A pilot clinical trial has been performed to identify the therapeutic effect of GlcNAc on IBD. Children with symptomatic Crohn's disease showed clear improvement after either oral or rectal administration of GlcNAc [118]. Therefore, GlcNAc shows promise as an inexpensive and non-toxic treatment for chronic IBD. Recently, a phase III clinical trial has just been finished [43].
3.4. Application of GlcNAc in cosmetics
Human skin contains stratum corneum and dermis to protect the body from harsh environmental conditions, such as dryness and UV irradiation. Stratum corneum plays a key role in maintaining the moisture and firmness of the skin. It consists of flattened dead cells, corneocytes and a complex of lipid matrix and natural moisture factors [119]. Dermis, the inner layer of the skin, is composed of collagen, elastin and GAGs, which are created by fibroblasts. Collagen is responsible for the resilience, strength, durability, elasticity, smoothness and plump appearance of healthy skin. Mucopolysaccharides, such as HA and proteoglycans, have a high water retention capacity and are important components in maintaining skin moisture.
When the amount of mucopolysaccharides decreases with age, the water retention and resilience of the skin decreases, and thereby causes rough skin and fine wrinkles. Cosmetics, commercially available topical preparations have been designed to increase the moisture content of the skin. Most cosmetics provide only a short-term replenishment of moisture-providing ingredients. Skin care ingredients that are derived from natural substances with high safety ratings are regarded as more beneficial.
HA, which is produced mainly by fibroblasts and keratinocytes, is a well known ingredient that holds water in the stratum corneum and dermis [120]. HA content is reported to decline with age, which may contribute to wrinkle formation and the decrease in elasticity of the skin [121]. HA is suggested to be a valuable ingredient in cosmetics. However, HA absorb inefficiently by either topical or oral administration due to its high molecular weight. Addition of GlcNAc, which is the building unit of HA, to cultured keratinocytes results in a dose-dependent increase in the production of HA [122], but has no effect on HA production of skin fibroblasts. Instead, GlcNAc enhances the proliferation and collagen expression of fibroblasts [123].
Topical administration of GlcNAc is used to improve skin quality. GlcNAc has been shown to be a good skin penetrant based on in vitro Franz cell testing. This result is consistent with the joint pain relieving effect of GlcNAc by topical administration [106]. In addition to its moisturizing effects, GlcNAc has been found to reduce the appearance of facial hyperpigmentation in an 8-week, double-blind clinical trial [124]. Using in vitro genomic experiments, the mechanism by which GlcNAc reduces melanin production has been found to involve the up-regulation of several genes, such as epidermal turnover genes and antioxidant-related genes, and the down-regulation of the cytoskeleton genes involved in melanosome transport. These changes are believed to be associated with pigment reduction [125]. Due to the versatile functions of GlcNAc, it is considered a valuable ingredient in cosmetics for improving skin wrinkles and color [126,127]. Because GlcNAc promotes the proliferation of keratinocytes and fibroblasts and increases the production of HA in the skin, it has also been successfully used to heal wounds [128,129].
3.5. GlcNAc is used as a substrate in sialic acid production
N-Acetylneuraminic acid (Neu5Ac) is the most common sialic acid and exists as >40 structural derivatives in mammalian and avian species [130]. Sialic acids are found at the distal ends of cell surface glycoconjugates and are major determinants of cellular recognition [131], serving as receptors for influenza viruses [131–133].
Infection of host cells by influenza viruses begins with the binding of hemagglutinin on the virus to the end of the sugar chain on the host cell. The infiltrating virus continuously proliferates in the cell. At the moment when the virus is to be released from the cell, the neuraminidase located on the surface of the virus hydrolyzes the binding site on the superficial layer of the host cell, and the freed virus binds to other non-infected cells. Novel pharmaceutical agents that are chemically synthesized from Neu5Ac to produce neuraminidase inhibitors are used to treat influenza infections. Indeed, Zanamivir, the guanidyl derivative of Neu5Ac, has been approved by several governments as an effective remedy for the treatment of the flu [132,133].
Because the Neu5Ac content in natural products is too low for the isolation of Neu5Ac with sufficient recovery and purity, novel processes had to be developed for the industrial production of this compound. Enzymatic synthesis of Neu5Ac from N-acetylmannosamine (ManNAc) and pyruvate using Neu5Ac aldolase as a catalyst has been reported. However, because ManNAc is very expensive, methods for the preparation of ManNAc from inexpensive GlcNAc, using chemical epimerization or enzymatic conversion by GlcNAc 2-epimerase, have been developed [134]. The simple and large-scale production of Neu5Ac using a two-step reaction was performed by Maru et al., in which 29 kg of Neu5Ac was obtained from 27 kg of GlcNAc, using recombinant GlcNAc 2-epimerase and Neu5Ac lyase as catalysts [135]. Lee et al. (2007) also developed a method to produce Neu5Ac using recombinant whole cells that express GlcNAc 2-epimerase and recombinant Neu5Ac lyase separately [136]. In summary, GlcNAc is an important substrate for the production of sialic acids.
3.6. Other applications of GlcNAc
A novel GlcNAc application has been developed by Xu et al. Because GlcNAc is a pure compound with a high level of safety, it is an appropriate candidate for multiple applications, especially for drug development. To test the special functions of GlcNAc, a novel bio-wave model was established with the use of Proteus mirabilis. In this model, the swarm colonies of P. mirabilis extend outward, and concentric rings are formed on the agar within 3 h as a result of the decay of metabolites [137]. In the presence of GlcNAc, many fine waves are formed in each ring, in addition to the typical concentric rings. Thus, GlcNAc was shown to portray a finer bio-wave characteristic than other compounds. This wave-promoting function may explain the special applications of GlcNAc described in Table 1. A series of cytological tests, animal tests and clinical trials have been performed to confirm the special functions of GlcNAc. Some of the successful cases have been filed as patents and are being utilized in the development of new drugs.
4. Conclusions
GlcNAc is prepared using chitin as a substrate by chemical, enzymatic and biotransformation methods. New methods using glucose as a substrate can also be applied to obtain GlcNAc using genetically modified microorganisms. Finally, the versatile functions of GlcNAc promote its use as a novel candidate for drug development.
This work was financially supported by National Science Council grant 98-2320-B-182-014-MY3 to C. R. Shen and 96-2221-E-131-008 as well as Ministry of Education Industry-Academy Cooperation grant 98B-38-006 to C. L. Liu. In addition, we appreciated American Journal Experts for the careful reading of the manuscript.
- Álvarez-Añorve, LI; Calcagno, ML; Plumbridge, J. Why Does Escherichia coli Grow More Slowly on Glucosamine Than on N-Acetylglucosamine? Effects of Enzyme Levels and Allosteric Activation of GlcN6P Deaminase (NagB) on Growth Rates. J. Bacteriol 2005, 187, 2974–2982, doi:10.1128/JB.187.9.2974-2982.2005. 15838023
- Chiu, PCN; Tsang, HY; Koistinen, H; Seppala, M; Lee, KF; Yeung, WSB. The Contribution of d-Mannose, l-Fucose, N-Acetylglucosamine, and Selectin Residues on the Binding of Glycodenlin Isoforms to Human Spermatozoa. Biol. Reprod 2004, 70, 1710–1719, doi:10.1095/biolreprod.103.023259. 14973265
- Promé, JC; Denarié, J; Truchet, G. Acylated Chitooligomers Are Molecular Signals That Mediate the Symbiotic Interactions Between Nitrogen-fixing Bacteria and Their Host Plants. Pure Appl. Chem 1998, 70, 55–60, doi:10.1351/pac199870010055.
- Oldroyd, GED; Mitra, RM; Wais, RJ; Long, SR. Evidence for Structurally Specific Negative Feedback in the Nod Factor Signal Transduction Pathway. Plant J 2001, 28, 191–199, doi:10.1046/j.1365-313X.2001.01149.x. 11722762
- Watkins, WM. Biochemistry and Genetics of the ABO, H, Lewis and P Blood Group Systems. Adv. Hum. Genet 1980, 10, 1–136. 6156588
- Izawa, M; Kumamoto, K; Mitsuoka, C; Kanamori, A; Ohmori, K; Ishida, H; Nakamura, S; Kurata-Miura, K; Sasaki, K; Nishi, T; Kannagi, R. Expression of Sialyl 6-Sulfo Lewis X Is Inversely Correlated with Conventional Sialyl Lewis X Expression in Human Colorectal Cancer. Cancer Res 2000, 60, 1410–1416. 10728707
- Kennedy, JF; White, CA. Bioactive Carbohydrates in Chemistry, Biochemistry and Biology; Ellis Horwood: Chichester, UK, 1983.
- D'Ambrosio, E; Casa, B; Bompani, R; Scali, G; Scali, M. Glucosamine Sulfate: a Controlled Clinical Investigation in Arthrosis. Pharmatherpeutica 1981, 2, 504–508.
- Laverty, S; Sandy, JD; Celeste, C; Vanchon, P; Marier, JF. Synovial Fluid Levels and Serum Pharmacokinetics in a Large Animal Model Following Treatment with Oral Glucosamine at Clinically Relevant Doses. Arthritis Rheum 2005, 52, 181–191, doi:10.1002/art.20762. 15641100
- Gooday, GW. The Ecology of Chitin Degradation. Adv. Microb. Ecol 1990, 11, 387–430.
- Hackman, RH; Goldberg, M. Light-scattering and Infrared-spectrophotometric Studies of Chitin and Chitin Derivatives. Carbohydr. Res 1974, 38, 35–45, doi:10.1016/S0008-6215(00)82336-8.
- Chitin and Chitinases; Jollès, P, Muzzarelli, RAA, Eds.; Birkhauser Verlag: Basel, Switzerland, 1999.
- Tanaka, T; Fujiwara, S; Nishikori, S; Fukui, T; Takagi, M; Imanaka, TA. Unique Chitinase with Dual Active Sites and Triple Substrate Binding Sites from the Hyperthermophilic Archaeon Pyrococcus kodakaraensis KOD1. Appl. Environ. Microbiol 1999, 65, 5338–5344. 10583986
- Ashry, ESHE; Aly, MRE. Synthesis and Biological Relevance of N-Acetylglucosamine-containing Oligosaccharides. Pure Appl. Chem 2007, 12, 2229–2242.
- DeAngelis, PL. Molecular Directionality of Polysaccharide Polymerization by the Pasteurella multocida Hyaluronan Synthase. J. Biol. Chem 1999, 274, 26557–26562, doi:10.1074/jbc.274.37.26557. 10473619
- Turely, EA; Bowman, P; Kytryk, MA. Effects of Hyaluronate and Hyaluronate Binding Proteins on Cell Motile and Contact Behaviour. J. Cell Sci 1985, 78, 133–145. 2419350
- Alho, AM; Underhill, CB. The Hyaluronate Receptor Is Preferentially Expressed on Proliferating Epithelial Cells. J. Cell Biol 1989, 108, 1557–1565, doi:10.1083/jcb.108.4.1557. 2466850
- Toole, BP; Jackson, G; Gross, J. Hyaluronate in Morphogenesis: Inhibition of Chondrogenesis in Vitro. Proc. Natl. Acad. Sci. USA 1972, 69, 1384–1386, doi:10.1073/pnas.69.6.1384. 4504347
- Brecht, M; Mayer, U; Schlosser, E; Prehm, P. Increased Hyaluronate Synthesis Is Required for Fibroblast Detachment and Mitosis. Biochem. J 1986, 239, 445–450. 3101667
- Dube, B; Luke, HJ; Aumailley, M; Prehm, P. Hyaluronan Reduces Migration and Proliferation in CHO Cells. Biochim. Biophys. Acta 2001, 1538, 283–289, doi:10.1016/S0167-4889(01)00079-9. 11336799
- Takaku, H; Ishida, HK; Fujita, M; Inazu, T; Ishida, H; Kiso, M. A Chemical Synthesis of GlcNAc[beta](1–4)GlcUA-UDP to Elucidate to Catalytic Mechanism of Hyaluronic Acid Synthesis (HAS). Synlett 2007, 5, 818–820.
- Aronson, NN; Kuranda, MJ. Lysosomal Degradation of Asn-Linked Glycoproteins. FASEB J 1989, 3, 2615–2622. 2531691
- Chien, LJ; Lee, CK. Hyaluronic Acid Production by Recombinant Lactococcus lactis. Appl. Microbiol. Biotechnol 2007, 77, 339–346, doi:10.1007/s00253-007-1153-z. 17805528
- Chien, LJ; Lee, CK. Enhanced Hyaluronic Acid Production in Bacillus subtilis by Coexpressing Bacterial Hemoglobin. Biotechnol. Prog 2007, 23, 1017–1022. 17691809
- Danishefsky, I; Steiner, H; Bella, A; Friedlander, A. Investigations on the Chemistry of Heparin: VI. Position of the Sulfate Ester Groups. J. Biol. Chem 1969, 244, 1741–1745. 5780839
- Bhavanandan, VP; Meyer, K. Mucopolysaccharides: N-Acetylglucosamine- and Galactose-6- sulfates from Keratosulfate. Science 1966, 151, 1404–1405, doi:10.1126/science.151.3716.1404. 4222407
- Scott, LJ; Clarke, NW; George, NJR; Shanks, JH; Testa, NG; Lang, SH. Interactions of Human Prostatic Epithelial Cells with Bone Marrow Endothelium: Binding and Invasion. Br. J. Cancer 2001, 84, 1417–1423, doi:10.1054/bjoc.2001.1804. 11355957
- Ugorski, M; Laskowska, A. Sialyl Lewis (a): a Tumor-Associated Carbohydrate Antigen Involved in Adhesion and Metastatic Potential of Cancer Cells. Acta Biochim. Pol 2002, 49, 303–311. 12362971
- Patel, KD; Cuvelier, SL; Wiehler, S. Selectins: Critical Mediators of Leukocyte Recruitment. Semin. Immunol 2002, 14, 73–81, doi:10.1006/smim.2001.0344. 11978079
- Parasadarao, NV; Wass, CA; Kim, KS. Endothelial Cell GlcNAc Beta 1–4GlcNAc Epitopes for Outer Membrane Protein A Enhance Traversal of Escherichia coli across the Blood-brain Barrier. Infect. Immun 1996, 64, 154–160. 8557333
- Aly, MRE; Rochaix, P; Amessou, M; Johannes, L; Florent, JC. Synthesis of Globo- and Isoglobotriosides Bearing a Cinnamoylphenyl Tag as Novel Electrophilic Thiol-Specific Carbohydrate Reagents. Carbohydr. Res 2006, 341, 2026–2036, doi:10.1016/j.carres.2006.03.044. 16777082
- Hartweck, LM; Scott, CL; Olszewski, NE. Two O-Linked N-Acetylglucosamine Transferase Genes of Arabidopsis thaliana L. Heynh. Have Overlapping Functions Necessary for Gamete and Seed Development. Genetics 2002, 161, 1279–1291. 12136030
- Holt, GD; Snow, CM; Senior, A; Haltiwanger, RS; Gerace, L; Hart, GW. Nuclear Pore Complex Glycoproteins Contain Cytoplasmically Disposed O-Linked N-Acetylglucosamine. J. Cell Biol 1987, 104, 1157–1164, doi:10.1083/jcb.104.5.1157. 3571327
- Schlesinger, PH; Rodman, JS; Doebber, TW; Stahl, PD; Lee, YC; Stowell, CP; Kuhlenschmidt, TB. The Role of Extra-hepatic Tissues in the Receptor-mediated Plasma Clearance of Glycoproteins Terminated by Mannose or N-Acetylglucosamine. Biochem. J 1980, 192, 597–606. 7236228
- Lord, JM. Precursors of Ricin and Ricinus communis Agglutinin. Eur. J. Biochem 1985, 146, 411–416, doi:10.1111/j.1432-1033.1985.tb08667.x. 3967664
- Liu, CL; Tsai, CC; Lin, SC; Wang, LI; Hsu, CI; Hwang, MJ; Lin, JY. Primary Structure and Function Analysis of the Abrus precatorius Agglutinin A Chain by Site-directed Mutagenesis. J. Biol. Chem 2000, 257, 1897–1901.
- Wu, AM; Wu, JH; Liu, JH; Chen, YY; Singha, B; Chow, LP; Lin, JY. Roles of Mammalian Structural Units, Ligand Cluster and Polyvalency in the Abrus precatorius Agglutinin and Glycoprotein Recognition Process. Mol. Immunol 2009, 16, 3427–3437.
- Spellman, MW; Basa, LJ; Leonard, CK; Chakel, JA; O'Connor, JV; Wilson, S; van Halbeek, H. Carbohydrate Structures of Human Tissue Plasminogen Activator Expressed in Chinese Hamster Ovary Cells. J. Biol. Chem 1989, 264, 14100–14111. 2503511
- Houpt, JB; McMillan, R; Wein, C; Paget-Dellio, SD. Effect of Glucosamine Hydrochloride in the Treatment of Pain of Osteoarthritis of the Knee. J. Rheumatol 1999, 26, 2423–243. 10555905
- Breidenbach, MA; Gallagher, JE; King, DS; Smart, BP; Wu, P; Bertozzi, CR. Targeted Metabolic Labeling of Yeast N-Glycans with Unnatural Sugars. Proc. Natl. Acad. Sci. USA 2010, 107, 3988–3993, doi:10.1073/pnas.0911247107. 20142501
- Shen, CR; Juang, JH; Tsai, ZT; Wu, ST; Tsai, FY; Wang, JJ; Liu, CL; Yen, TC. Preparation, Characterization and Application of Superparamagnetic Iron Oxide Encapsulated with N-[(2-Hydroxy-3-trimethylammonium) propyl] Chitosan Chloride. Carbohydr. Polym 2010, doi:10.1016/j.carbpol.2010.07.067.
- Levin, RM; Krieger, NN; Winzler, RJ. Glucosamine and Acetylglucosamine Tolerance in Man. J. Lab. Clin. Med 1961, 58, 927–932. 14464574
- Liu, Y; Li, Z; Liu, G; Jia, J; Li, S; Yu, C. Liquid Chromatography–Tandem Mass Spectrometry Method for Determination of N-Acetylglucosamine Concentration in Human Plasma. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci 2008, 862, 150–154, doi:10.1016/j.jchromb.2007.11.043. 18165162
- Kobata, A; Ginsburg, V. Oligosaccharides of Human Milk: I. Isolation and Characterization. Arch. Biochem. Biophys 1969, 130, 509–513, doi:10.1016/0003-9861(69)90063-0. 5778662
- Miller, JB; Bull, S; Miller, J; McVeagh, P. The Oligosaccharide Composition of Human Milk: Temporal and Individual Variations in Monosaccharide Components. J. Pediatri. Gastroenterol. Nutr 1994, 19, 371–376, doi:10.1097/00005176-199411000-00001.
- Bakkers, J. An Important Developmental Role for Oligosaccharides during Early Embryogenesis of Cyprinid Fish. Proc. Natl. Acad. Sci. USA 1997, 94, 7982–7986, doi:10.1073/pnas.94.15.7982. 9223299
- Tharanathan, RN; Kittur, FS. Chitin—the Undisputed Biomolecule of Great Potential. Crit. Rev. Food Sci. Nutr 2003, 43, 61–87, doi:10.1080/10408690390826455. 12587986
- Howard, MB; Ekborg, NA; Weiner, RM; Hutcheson, SW. Detection and Characterization of Chitinases and Other Chitin-modifying Enzymes. J. Ind. Microbiol. Biotechnol 2003, 30, 627–635, doi:10.1007/s10295-003-0096-3. 14610656
- Kurita, K. Chitin and Chitosan: Functional Biopolymers from Marine Crustaceans. Mar. Biotechnol 2006, 8, 203–226, doi:10.1007/s10126-005-0097-5. 16532368
- Bohlman, JA; Schisler, DO; Hwang, KO; Hennling, JP; Trinkle, JR; Anderson, TB; Steinke, JD; Vanderhoff, A. N-Acetyl-d-glucosamine and Process for Producing N-Acetyl-d-glucosamine. US Patent 6693188B2, 2004.
- Sashiwa, H; Fujishima, S; Yamano, N; Kawasaki, N; Nakayama, A; Muraki, E; Aiba, S. Production of N-Acetyl-d-glucosamine from β-Chitin by Enzymatic Hydrolysis. Chem. Lett 2001, 31, 308–309.
- Roseman, S; Ludoweig, J. N-Acetylation of Hexosamines. J. Am. Chem. Soc 1954, 76, 301–302, doi:10.1021/ja01630a098.
- Kohn, P; Winzler, RJ; Hoffmann, RC. Metabolism of d-Glucosamine and N-Acetyl-d-glucosamine in the Intact Rat. J. Biol. Chem 1962, 237, 304–308. 14457848
- Zhan, WS. Process for Preparing Refined N-Acetyl-d-aminoglucose. CN Patent 1907993, 2007.
- Ryosuke, K; Yoshiharu, M; Kazuaki, K; Kazuo, S. Production of Natural-type N-Acetyl-d-glucosamine. JP Patent 200281696, 2002.
- Seo, S; King, JM; Prinyawiwatkul, W. Simultaneous Depolymerization and Decolorization of Chitosan by Ozone Treatment. J. Food Sci 2007, 72, C522–C526, doi:10.1111/j.1750-3841.2007.00563.x. 18034714
- Gooday, BW. Biosynthesis of the Fungal Wall—Mechanisms and Implications. The First Fleming Lecture. J. Gen. Microbiol 1977, 99, 1–11. 864434
- Fuchs, RL; McPherson, SA; Drahos, DJ. Cloning of a Serratia marcescens Gene Encoding Chitinase. Appl. Environ. Microbiol 1986, 51, 504–509. 16347012
- Abeles, FB; Bosshart, RP; Forrence, LE; Habig, WH. Preparation and Purification of Glucanase and Chitinase from Bean Leaves. Plant Physiol 1971, 47, 129–134, doi:10.1104/pp.47.1.129. 16657567
- Cohen-Kupiec, R; Chet, I. The Molecular Biology of Chitin Digestion. Curr. Opin. Biotechnol 1998, 9, 270–277, doi:10.1016/S0958-1669(98)80058-X. 9650272
- Zhu, Z; Zheng, T; Homer, RJ; Kim, YK; Chen, NY; Cohn, L; Hamid, Q; Elias, JA. Acidic Mammalian Chitinase in Asthmatic Th2 Inflammation and IL-13 Pathway Activation. Science 2004, 304, 1678–1682, doi:10.1126/science.1095336. 15192232
- Yang, CJ; Liu, YK; Liu, CL; Shen, CN; Kuo, ML; Su, CC; Tseng, CP; Yen, TC; Shen, CR. Inhibition of Acidic Mammalian Chitinase by RNA Interference Suppresses OVA-Sensitized Allergic Asthma. Hum. Gene Ther 2009, 20, 1597–1606, doi:10.1089/hum.2008.092. 19548841
- Cohen-Kupiec, R; Chet, I. The Molecular Biology of Chitin Digestion. Curr. Opin. Biotechnol 1998, 9, 270–277, doi:10.1016/S0958-1669(98)80058-X. 9650272
- Lee, EA; Pan, CH; Son, JM; Kim, SI. Isolation and Characterization of Basic Exochitinase from Leaf Extract of Rehmannia glutinosa. Biosci. Biotechnol. Biochem 1999, 63, 1781–1783, doi:10.1271/bbb.63.1781.
- Mori, T; Ichikawa, W; Kita, Y; Tetsuka, Y. Method for Fermentative Production of N-Acetyl- d-glucosamine by Microorganism. US Patent 20100055746, 2010.
- Roberts, RL; Cabib, E. Serratia marcescens Chitinase: One-step Purification and Use for the Determination of Chitin. Anal. Biochem 1982, 127, 402–412, doi:10.1016/0003-2697(82)90194-4. 6762111
- Oppenheim, AB; Chet, I. Cloned Chitinase in Fungal Plant—Pathogen Control Strategies. Trends Biotechnol 1992, 10, 392–394, doi:10.1016/0167-7799(92)90281-Y.
- Pan, CH; Rim, SL; Kim, SI. Expression of Two cDNAs Encoding Class I Chitinases of Rice in Escherichia coli. Biosci. Biotech. Biochem 1996, 60, 1346–1348, doi:10.1271/bbb.60.1346.
- Yalpani, M; Pantaleone, D. An Examination of the Unusual Susceptibilities of Aminoglycans to Enzymatic Hydrolysis. Carbohydr. Res 1994, 256, 159–175, doi:10.1016/0008-6215(94)84235-3.
- Muzarelli, RAA. Depolymerization of Chitins and Chitosans with Hemicellulase, Lysozyme, Papain and Lipase. In Chitin Handbook; Muzzarelli, RAA, Peter, MG, Eds.; Atec: Grottammare, Italy, 1997; pp. 153–163.
- Aiba, S; Muraki, E. Preparation of Higher N-Acetylchitooligosaccharides in High Yields. In Advance in Chitin Science; Chen, RH, Chen, HC, Eds.; Rita Advertising Co. Ltd: Taipei, Taiwan, 1999; Volume III, pp. 89–96.
- Sashiwa, H; Fujishima, S; Yamano, N; Kawasaki, N; Nakayama, A; Muraki, E; Hiraga, K; Oda, K; Aiba, S. Production of N-Acetyl-d-glucosamine from α-Chitin by Crude Enzymes from Aeromonas hydrophila H2330. Carbohydr. Res 2002, 337, 761–763, doi:10.1016/S0008-6215(02)00034-4. 11950472
- Pichyangkura, R; Kudan, S; Kuttiyawang, K; Sukwattanasinitt, M; Aiba, S. Quantitative Production of 2-Acetoamodo-2-d-glucose from Crystalline Chitin by Bacterial Chitinase. Carbohydr. Res 2002, 337, 557–559, doi:10.1016/S0008-6215(02)00007-1. 11890893
- Sashiwa, H; Fujishima, S; Yamano, N; Kawasaki, N; Nakayama, A; Muraki, E; Sukwattanasinitt, M; Pichyangkura, R; Aiba, S. Enzymatic Production of N-Acetyl-d-glucosamine from Chitin. Degradation Study of N-Acetylchitooligosaccharide and the Effect of Mixing of Crude Enzymes. Carbohydr. Polym 2003, 51, 391–395, doi:10.1016/S0144-8617(02)00192-3.
- Rinaudo, MA. Chitin and Chitosan: Properties and Applications. Prog. Polym. Sci 2006, 31, 603–632, doi:10.1016/j.progpolymsci.2006.06.001.
- Ilankovan, P; Hein, S; Ng, C; Trung, TS; Stevens, WF. Production of N-Acetyl Chitobiose from Various Chitin Substances Using Commercial Enzymes. Carbohydr. Polym 2006, 63, 245–250, doi:10.1016/j.carbpol.2005.08.060.
- Kurita, K; Kaji, Y; Mori, T; Nishiyama, Y. Enzymatic Degradation of β-Chitin: Susceptibility and the Influence of Deacetylation. Carbohydr. Polym 2003, 42, 19–21.
- Shen, CR; Chen, YS; Yang, CJ; Chen, JK; Liu, CL. Colloid Chitin Azure Is a Dispersible, Low-cost, Substrate for Chitinase Measurements in a Sensitive, Fast, Reproducible, Assay. J. Biomol. Screen 2010, 15, 213–217, doi:10.1177/1087057109355057. 20042532
- Kawasaki, IK; Morita, T. Enzymatic Decomposition of Chitin Containing Materials. US Patent 5262310, 1993.
- Immanaka, T; Fukui, T; Fujiwara, S. Chitinase from Thermococcus kodakaraensis KOD1. Methods Enzymol 2001, 330, 319–329. 11210511
- Guo, SH; Chen, JK; Lee, WC. Purification and Characterization of Extracellular Chitinase from Aeromonas schubertii. Enzyme Microb. Technol 2004, 35, 550–556, doi:10.1016/j.enzmictec.2004.08.025.
- Liu, CL; Shen, CR; Hsu, FF; Chen, JK; Wu, PT; Guo, SH; Lee, WC; Yu, FW; Mackey, ZB; Turk, J; Gross, ML. Isolation and Identification of Two Novel SDS-Resistant Secreted Chitinases from Aeromonas schubertii. Biotechnol. Prog 2009, 25, 124–134, doi:10.1002/btpr.100. 19197977
- Chen, MH; Chan, HY; Wu, CL; Chuang, SH; Hwang, IE; Chen, YL; Yuan, GF. Production of Chitin and Chitosan. US Patent 6255085 B1, 2001.
- Barnhill, JG; Fye, CL; Reda, DJ; Harris, CL; Clegg, DO. Is All Glucosamine Alike? Clarifying the Controversies for Product Selection and Clinical Research. J. Compl. Integr. Med 2009, 6, 17.
- Raetz, E; Leuba, JL; Di Giambattista, R; Federici, F; Fenice, M. Chitinolytic Enzymes Production by Penicillium janthinellum. EP Patent 0885954 A1, 1998.
- Louise, CA; Pedro, A; Charles, AH. Process for Producing N-Acetyl-d-glucosamine. US Patent 5998173, 1999.
- Li, YL; Wu, ST; Yu, ST; Too, JR. Screening of a Microbe to Degrade Chitin. Taiwanese J. Agric. Chem. Food Sci 2005, 43, 410–418. (Chinese).
- Chern, LL; Stackebrandt, E; Lee, SF; Lee, FL; Chen, JK; Fu, HM. Chitinibacter tainanensis Gen. nov. sp. nov., a Chitin-degrading Aerobe from Soil in Taiwan. Int. J. Syst. Evol. Microbiol 2004, 54, 1387–1391, doi:10.1099/ijs.0.02834-0. 15280319
- Chen, JK; Shen, CR; Fang, BS; Huang, TL; Liu, CL. The N-Acetyl-glucosamine Obtained from Chitin with Chitinibacter tainanensis. Carbohydr. Polym 2010. submitted.
- Doi, RH; Kosugi, A; Murashima, K; Tamura, Y; Han, SO. Cellulosomes from Mesophilic Bacteria. J. Bacteriol 2003, 185, 5907–5914, doi:10.1128/JB.185.20.5907-5914.2003. 14526000
- Deng, MD; Angerer, JD; Cyron, D; Grund, AD; Jerrell, TA, Jr; Leanna, C; Mathre, O; Rossen, R; Running, J; Severson, D. Process and Materials for Producing of Glucosamine and N-Acetylglucosamine. KR Patent 20050053534, 2005.
- Yoon, SH; Rhee, JS. Lipid from Yeast Fermentation: Effects of Cultural Conditions on Lipid Production and Its Characteristics of Rhodotorula glutinis. J. Am. Oil Chem. Soc 1983, 60, 1281–1286, doi:10.1007/BF02702101.
- Hamme, JV; Singh, A; Ward, O. Physiological Aspects Part 1 in a Series of Papers Devoted to Surfactants in Microbiology and Biotechnology. Biotechnol. Adv 2006, 24, 604–620, doi:10.1016/j.biotechadv.2006.08.001. 16979315
- Wendland, J; Schaub, Y; Walther, A. N-Acetylglucosamine Utilization by Saccharomyces cerevisiae Based on Expression of Candida albicans NAG Genes. Appl. Environ. Microbiol 2009, 75, 5840–5845, doi:10.1128/AEM.00053-09. 19648376
- Vidal, Y; Plana, PR; Bizzari, D; Rovati, A. Articular Cartilage Pharmacology: I. In Vitro Studies on Glucosamine and Non-Steroidal Anti-inflammatory Drugs. Pharmacol. Res. Commun 1978, 10, 557–569, doi:10.1016/S0031-6989(78)80054-X. 151283
- Heim, HK; Oestmann, A; Thiele, H; Sewing, KH. Incorporation of N-Acetyl-[14C]d-glucosamine and [3H]l-Leucine by Isolated Pig Gastric Mucosal Cells. Digestion 1989, 44, 26–35, doi:10.1159/000199889. 2599280
- Shoji, A; Iga, T; Inagaki, S; Kobayashi, K; Matahira, Y; Sakai, K. Metabolic Disposition of [14C] N-Acetylglucosamine in Rats. Chitin Chitosan Res 1999, 5, 34–42.
- Lee, KY; Shibutani, M; Takagi, H; Arimura, T; Takigami, S; Uneyama, C; Kato, N; Hirose, M. Subchronic Toxicity Study of Dietary N-Acetylglucosamine in F344 Rats. Food Chem. Toxicol 2004, 42, 687–695, doi:10.1016/j.fct.2003.12.005. 15019194
- Xu, Q; Liu, J. Use of N-Acetyl d-glucosamine in Treatment of Organ Lesion Related to Toxicosis of Drugs or Chemicals. US Patent 2006281707, 2006.
- Felson, DT. Epidemiology of Hip and Knee Osteoarthritis. Epidemiol. Rev 1988, 10, 1–28. 3066625
- Creamer, P. Osteoarthritis Pain and Its Treatment. Curr. Opin. Rheumatol 2000, 12, 450–455, doi:10.1097/00002281-200009000-00019. 10990186
- Rovati, L; Casula, P; Mascherpa, S. N-Acetylglucosamine for Treating Degenerative Afflictions of the Joints. US Patent 3697652, 1972.
- Talent, JM; Gracy, RW. Pilot Study of Oral Polymeric N-Acetyl-d-glucosamine as a Potential Treatment for Patients with Osteoarthritis. Clin. Ther 1996, 18, 1184–1190, doi:10.1016/S0149-2918(96)80073-7. 9001835
- Chrisope, GL; Rose, R. Product and Method for Treating Joint Disorders in Vertebrates. US Patent 6344220, 1999.
- Tamai, Y; Miyatake, K; Okamoto, Y; Takamori, Y; Sakamoto, K; Minami, S. Enhanced Healing of Cartilaginous Injuries by N-Acetyl-d-glucosamine and Glucuronic Acid. Carbohydr. Polym 2003, 54, 251–262, doi:10.1016/S0144-8617(03)00170-X.
- Wilkie, D. Topical Composition for the Treatment of Joint Damage or Pain. GB Patent 2403405, 2005.
- Marcum, FD; Seanor, JW. Composition and Method for Treating Rheumatoid Arthritis. US Patent 2008003258, 2007.
- Kamel, M; Hanafi, N; Bassiouni, M. Inhibition of Elastase Enzyme Release from Human Polymorphonuclear Leukocytes by N-Acetylgalactosamine and N-Acetylglucosamine. Clin. Exp. Rheumatol 1991, 9, 17–21. 2054963
- Kamel, M; Alnahdi, M. Inhibition of Superoxide Anion Release from Human Polymorphonuclear Leukocytes by N-Acetylgalactosamine and N-Acetylglucosamine. Clin. Rheumatol 1992, 11, 254–260, doi:10.1007/BF02207968. 1319880
- Murch, SH; Braegger, CP; Walker-Smith, JA; McDonald, TT. Localization of Tumor Necrosis Factor α by Immunohistochemistry in Chronic Inflammatory Bowel Disease. Gut 1993, 34, 1705–1709, doi:10.1136/gut.34.12.1705. 8031350
- Kanazawa, M; Fukudo, S. Effects of Fasting Therapy on Irritable Bowel Syndrome. Int. J. Behav. Med 2006, 13, 214–220, doi:10.1207/s15327558ijbm1303_4. 17078771
- Monnikes, H; Schmidtmann, M; van der Voort, IR. Drug Therapy for Irritable Bowel Syndrome. What Works, What Doesn't Work and for Whom? Internist (Berl) 2006, 47.
- Burton, AF; Gislason, S. Methods and Composition for Treatment of Food Allergy. US Patent 5192750, 1993.
- Burton, AF; Mclean, DI. Method and Composition for Treating Psoriasis. US Patent 5217962, 1993.
- Karzed, K; Domenjoz, R. Effects of Hexosamine Derivatives and Uronic Acid Derivatives on Glycosaminoglycan Metabolism of Fibroblast Cultures. Pharmocology 1971, 5, 337–345.
- Burtan, AF. N-Acetyl Glucosamine as a Cytoprotective Agent. KR Patent 0145715 B, 1998.
- Burtan, AF; Freeman, HJ. N-Acetyl Glucosamine as a Gastroprotective Agent. WO Patent 9323055, 1993.
- Salvatore, S; Heuschkel, R; Tomlin, S; Davies, SE; Edwards, S; Walker-Smith, JA; French, I; Murch, SH. A Pilot Study of N-Acetyl Glucosamine, a Nutritional Substrate for Glycosaminoglycan Synthesis, in Paediatric Chronic Inflammatory Bowel Disease. Aliment. Pharmacol. Ther 2000, 14, 1567–1579, doi:10.1046/j.1365-2036.2000.00883.x. 11121904
- Wertz, PW; van der Bergh, B. The Physical, Chemical and Functional Properties of Lipid in the Skin and Other Biological Barrier. Chem. Phys. Lipids 1998, 91, 85–96, doi:10.1016/S0009-3084(97)00108-4. 9569614
- Laurent, TC; Fraser, JR. Hyaluronan. FASEB J 1992, 6, 2397–2404. 1563592
- Ghersetich, I; Lotti, T; Campanile, G; Grappone, C; Dini, G. Hyaluronic Acid in Cutaneous Intrinsic Agent. Int. J. Dermatol 1994, 33, 119–122, doi:10.1111/j.1365-4362.1994.tb01540.x. 8157393
- Sayo, T; Sakai, S; Inoue, S. Synergestic Effect of N-Acetylglucosamine and Retinoids on Hyaluronan Production in Human Keratinocytes. Skin Pharmacol. Physiol 2004, 17, 77–83, doi:10.1159/000076017. 14976384
- Chen, RH; Hsu, CN; Chung, MY; Tsai, WL; Liu, CH. Effect of Different Concentrations of Collagen, Ceramides, N-Acetyl Glucosamine, or Their Mixture on Enhancing the Proliferation of Keratinocytes, Fibroblasts and the Secretion of Collagen and/or the Expression of mRNA of Type I Collagen. J. Food Drug Anal 2008, 16, 66–74.
- Bissett, D; Robinson, LR; Raleigh, PS; Miyamoto, K; Hakozaki, T; Li, J; Klem, GR. Reduction in the Appearance of Facial Hyperpigmentation by Topical N-Acetyl Glucosamine. J. Cosmet. Dermatol 2007, 6, 20–26, doi:10.1111/j.1473-2165.2007.00295.x. 17348991
- Bissett, D; Farmer, T; McPhail, S; Reichling, T; Tiesman, JP; Juhlim, KD; Hurley, GJ; Robinson, MK. Genomic Expression Changes Induced by Topical N-Acetylglucosamine in Skin Equivalent Cultures in Vitro. J. Cosmet. Dermatol 2007, 6, 232–238, doi:10.1111/j.1473-2165.2007.00339.x. 18047607
- Riordan, NH. Skin Treatment System. US Patent 5866142, 1999.
- Hwang, JI; Kim, KS. Cosmetics Composition Comprising Extract of Natural Materials for Improving Acne and Skin Wrinkles and Whitening Skin. KR Patent 20050004355 A, 2005.
- Minami, S; Okamoto, Y. Drug for Remedy or Treatment of Wound. EP Patent 1749532, 2007.
- Flessner, MF; Lofthouse, J; Williams, A. Chronic Alteration of Sub-Peritoneal Tissue and Peritoneal Transport. Adv. Perit. Dial 2002, 18, 12–14. 12402578
- Vimr, E; Lichtensteiger, C. To Sialylate, or not to Sialylate: That Is the Question. Trends Microbiol 2002, 10, 254–257, doi:10.1016/S0966-842X(02)02361-2. 12088651
- Tanner, ME. The Enzymes of Sialic Acid Biosynthesis. Bioorg. Chem 2005, 33, 216–228, doi:10.1016/j.bioorg.2005.01.005. 15888312
- Maru, I; Ohnishi, J; Ohta, Y; Tsukada, Y. Why Is Sialic Acid Attracting Interest Now? Complete Enzymatic Synthesis of Sialic Acid with N-Acetylglucosamine 2-Epimerase. J. Biosci. Bioeng 2002, 93, 258–265. 16233198
- Gubareva, LV; Kaiser, L; Hayden, FG. Influenza Virus Neuraminidase Inhibitors. Lancet 2000, 827–835.
- Tabata, K; Koizumi, S; Endo, T; Ozaki, A. Production of N-Acetyl-d-neuraminic Acid by Coupling Bacteria Expressing N-Acetyl-d-glucosamine 2-Epimerase and N-Acetyl-d-neuraminic Acid Synthetase. Enzyme Microb. Technol 2002, 30, 327–333, doi:10.1016/S0141-0229(01)00515-4.
- Maru, I; Ohnishi, J; Ohta, Y; Tsukada, T. Simple and Large-Scale Production of N-Acetyl-neuraminic Acid from N-Acetyl-d-glucosamine and Pyruvate Using N-Acetyl-d-glucosamine 2- Epimerase and N-Acetyl-neuraminic Acid Lyase. Carbohydr. Res 1998, 306, 575–578, doi:10.1016/S0008-6215(97)10106-9. 9679278
- Lee, YC; Chien, HR; Hsu, WH. Production of N-Acetyl-neuraminic Acid by Recombinant Whole Cells Expressing Anabaena sp. CH1 N-Acetyl-d-glucosamine 2-epimerase and Escherichia coli N-Acetyl-neuraminic Acid Lyase. J. Biotechnol 2007, 129, 453–460, doi:10.1016/j.jbiotec.2007.01.027. 17349707
- Rauprich, O; Matsushita, M; Weijer, CJ; Siegert, F; Esipov, SE; Shapiro, JA. Periodic Phenomena in Proteus mirabilis Swarm Colony Development. J. Bacteriol 1996, 178, 6525–6538. 8932309
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in Preparation of Drugs for the Treatment of Cancer and Metastasis. EP Patent 1666046 A1, 2006.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in Treatment of Local Lesions or Systematic Symptoms Related to Autoimmune Reactions. EP Patent 1611896 A1, 2006.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine for Preparing Medicines for Urogenital Tract Infection's Treatment and Prevention. US Patent 2006/0142243 A1, 2006.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in Treatment of Local Lesions and Systematic Symptoms Related to Infections of Virus and Bacteria. US Patent 2007/0042995 A1, 2007.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in the Manufacture of Pharmaceutical Useful for Preventing and Treating Sexual Disorder. US Patent 7015207 B2, 2006.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in Treatment of Non-Specific Infections Related to Physical or Chemical Factors. EP Patent 1611894, 2006.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in the Manufacture of Pharmaceutical Useful for Adjuvant Treatment of Perianal Disease. US Patent 2005119224, 2005.
- Xu, Q; Liu, J; Yuan, Z. Compound Antibacterial Drugs Comprising N-Acetyl-d-glucosamine. US Patent 20070191291, 2007.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in the Manufacture of Pharmaceutical Useful for Treating Cervical Erosion. US Patent 2004138174 A1, 2004.
- Xu, Q. Application of N-Aceto-d-aminoglucose in Medicinal Preparation for Curing Respiratory Tract Disease. CN Patent 1156026, 1997.
- Xu, Q; Liu, J. Use of N-Acetyl-d-glucosamine in Treatment of Organ Lesions Related to Toxicosis of Drugs or Chemicals. US Patent 7345030 B2, 2008.
- Xu, Q; Liu, J; Yuan, Z. Antibacterial Compositions of N-Acetyl-d-aminoglycosamine and Antibiotics. EP Patent 1669077, 2006.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in the Manufacture of Pharmaceutical Useful for Suppress Side Effect of Radiotherapy and Chemotherapy. US Patent 7037904 B2, 2006.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in the Manufacture of Pharmaceutical Useful for Treating Cardio-Cerebrovascular Anoxemia. US Patent 7074774 B2, 2006.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in the Manufacture of Pharmaceutical Useful for Treating Motion Sickness. US Patent 6946452 B2, 2005.
- Xu, Q; Yuan, Z; Liu, J. Use of N-Acetyl-d-glucosamine in Preparation of Drugs for Modulating Microorganisms on Mucus Membrane. CA Patent 2539286 A1, 2005.
- Xu, Q. Application of N-Acetyl-d-aminoglucose for Preparing Skin Sanitary Article Preparation. CN Patent 1156028, 1997.
- Xu, Q; Liu, J; Yuan, Z. Use of N-Acetyl-d-glucosamine in Preparation of Skin Membrane Mucosa Microecological Balance Regulator Medicines. CN Patent 1470244, 2004.
- Xu, Q; Liu, J; Yuan, Z. The Use of N-Acetyl Glucosamine as Additive in Milk Products. WO Patent 2004093556, 2004.
- Xu, Q; Liu, J; Yuan, Z. The Use of N-Acetyl Glucosamine as Additive in Beer and Beer Comprising N-Acetyl Glucosamine. WO Patent 2004085603, 2004.
- Xu, Q; Liu, J; Yuan, Z. The Use of N-Acetyl Glucosamine as Additive in Wine and Wine Comprising N-Acetyl Glucosamine. WO Patent 2004085604, 2004.
|Table 1. Applications of GlcNAc derived from the bio-wave model
|Disease Treatment
|cancer and metastasis
|animal, human
|autoimmune reactions
|cell, animal, human
|urogenital tract infection
|cell, human
|viral or bacterial infection
|cell, animal
|sexual disorder
|non-specific inflammation
|animal, human
|perianal disease
|cell, animal, human
|intestinal disease
|cervical erosion
|bacteria, human
|respiratory tract disease
|Disease Prevention
|toxicosis of drugs or chemicals
|cell, animal
|side effects of radiotherapy and chemotherapy
|cardio-cerebrovascular anoxemia
|motion sickness
|modulation of microorganisms on mucus membrane
|animal, human
|skin sanitary article preparation
|microecological balance of mucosa
|bacteria, human
|Food supplement
|additive in milk
|animal, human
|additive in beer
|animal, human
|additive in wine
|animal, human
© 2010 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).


text:
How Did Twitter React To The Birth Of Jay-Z And Beyonce's Baby? [STUDY]
Back in August, when Beyonce Knowles first announced her pregnancy to the world at the MTV Video Music Awards, the event triggered a new tweets per second (TPS) record on Twitter – a mark which stood until December of last year.
Baby Blue Ivy Carter was born on January 7, and while Beyonce and husband Jay-Z have yet to give the world a glimpse of the newborn, Twitter reacted to the news with considerable gusto.
Social analytics company Simply Measured tracked the impact of the birth of Blue Ivy on Twitter, and while the story didn't reach the heights of Beyonce's VMA announcement, things did get pretty wild.
The spike, which peaked at 2,379 TPS on Saturday at 11.15pm PST, was helped by the usual assortment of celebrities and other Twitter power-users sending their well-wishes to the new parents. Hip hop mogul Russell Simmons got the ball rolling, and as Ryan Seacrest, Perez Hilton, People magazine and others waded in, the news rapidly made its way around Twitter.
- 9 Risky Overshares From Twitter Celebrities
- Ellen's Oscar Selfie Is Now A Piece Of Art At Twitter HQ
- George Clooney: I Don't Tweet Because I Like To Have A Drink At Night
- Celebrities And Fans Pay Tribute To Uncle Phil On Twitter


text:
This hearty beef chili features boneless chuck roast and a medley of traditional chili seasonings. Serve with homemade cornbread for a complete meal.
Southern Living OCTOBER 2005
Brown meat, in batches, in a Dutch oven over medium-high heat. Remove meat, reserving drippings in Dutch oven. Add chili powder to Dutch oven; cook, stirring constantly, 2 minutes. Stir in tomato paste; cook 5 minutes.
Return beef to Dutch oven. Stir in beef broth and next 9 ingredients; bring to a boil. Reduce heat to low, and simmer, uncovered, stirring occasionally, 1 1/2 hours or until beef is tender. Serve with cornbread sticks, if desired, and desired toppings.
If you've had spices longer than a year, it might be time to replace them. Seasonings tend to dull in flavor the longer they sit on the shelf. Always store them in a cool, dry place.
Go to Full Version of


text:
Randam Ron Paulism
War is not popular. It may seem popular in the short run, when there appears to be an immediate victory and everyone is gloating but war is not popular. War is not popular. People get killed, and body bags end up coming back. War is very unpopular, and it is not the politically smart thing to do"
—Ron Paul
ENGLISH Alex Jones: My father is Jewish; WNs destroy but charge …
Hopelessly genuine white people find it intolerable that Adolf Hitler had "storm troopers."
But there was a really good reason, an uncommonly unsentimental reason for Hitler combining a charge troopers:
Their sign was:
Willst du nicht mein Bruder sein, so schlage ich dir basement Schädel ein.
("If we will not my hermit be, we will pound your skull in three.')
The One People's Project goons are a Reds of today. They have assaulted pacific white tellurian rights activists adult and down a Northeast.
Not prolonged ago, they invaded a David Irving assembly in Manhattan and totally pennyless adult a long-planned assembly a 70-year-old historian had organized, spraying rip gas in a face of several attendees, including Irving's really appealing immature womanlike assistant.
The military and FBI, confronted with transparent justification of terrorism and swindling to repudiate polite rights. immediately did…….
Emboldened by a LACK OF STORM TROOPERS, a fat black faggot Darryl Lamont Jenkins has continued his terrorism, recently shutting down a American Renaissance conference. His supporters finished militant genocide threats opposite any hotel that had resolved to horde Jared Taylor's group.
Here is a latest hate-filled conflict on a leisure of debate of white Americans in their possess republic by revolutionary extremist psychopathic bullies.
Roy Beck, a fair-haired, aging German-American, is a executive of NumbersUSA ("for reduce immigration levels"), and his commendable organisation orderly a counter-demonstration yesterday to a vast Hispanic pro-amnesty proof on a Washington DC Mall.
You only HAVE to see this video clip.
There he is, aging aged white male Roy. beleaguered by non-whites and paleface psychopaths who are younger than he is.
(Believe me, a Hispanics roar any day how WE are aging and they are immature and fit. we get hatred emails, from a protected electronic distance, job me an "old man" constantly from Hispanics.)
Now demeanour during Beck's "protection": BLACK bodyguards….. Where are a WHITE MEN?
Observe Beck, whose IQ is substantially 50% aloft than a morons, enchanting them in a sum non-debate where they only scream slogans during him…..
The (in THEIR definition of a word) nazi nazi racist Hispanics are doing all only brief of assault to close down his leisure of expression, and anticipating that underneath impassioned irritation HE will get aroused so a black DC cops can detain HIM.
They are floating whistles and popping ballons so as to expostulate him crazy, and holding signs in an annoying demeanour to retard his possess camerawoman.
Without any question, a ju lerned these scum.
A white revolutionary lady harasses Roy as a antagonistic "mime." She is roughly positively nonetheless another crazy Irish-American. All we have to do is tell an Irishman that anyone is an "underdog" and he or she will be out their shedding his blood for that organisation — blax, juze, queers and mex — all groups that HATE white people such as a Irish! This "mime" (God, we and many people hate mimes, and Marcel Marceau, a "famous mime," was a ju.) This idiot does all probable to confuse viewers from Beck as he is being interviewed and is creation logic and convincing points.
But how can all THAT be fair?
It ain't — though a psychopath LAUGHS during your right to speak!
He has a right to HATE and a right to BULLY — since we are diseased and he is A BRUTAL SADIST.
One low-IQ Latina keeps screaming to Beck that a Hispanics will mount together. Well then, since should a whites not??
And of march she trots out a bad-science logic since she presumably has a right to be in OUR country:
"We were here prolonged before you."
And this is since they fear my Solutrean truth, since we proclaim, we did so on a Discovery Channel, and will do so in a approach that a white masses GET, that WE WHITES were here TEN THOUSAND YEARS before a Injuns (and Mexicans are only Injuns with a smidgen of both white and also negro blood).
And that it was THEIR ancestors who genocided US.
That is since a Solutrean dwindle has a stalk — since we are a spear-minded people. And a Solutrean sign is:
REMEMBER. And thus, never again.
The Dinner Roll ..
I went to a White House for a private cooking with a President.
I am a reputable businessman, with a bureau that produces memory chips for computers and unstable electronics.
There was some speak that my attention was being scrutinized by a administration, though we paid it no mind. we live in a FREE country. The supervision won't worry me if I've damaged no laws. My resources was EARNED honestly, and dining with a President is an honor.
I checked my coat, was greeted by a Chief of Staff, and assimilated a President in a yellow dining room.
We sat opposite from any other during a list draped in white linen. The Great Seal was embossed on a china. Uniformed staff served a dinner.
The dish was served, and we was dismayed when my waiter unexpected reached out, plucked a cooking hurl off my image and began roaming it as he walked behind to a kitchen..
"Sorry 'bout that," pronounced a President. "Andrew is really hungry."
"I don't appreciate…" we began, though as we looked into a ease brownish-red eyes opposite from me, we felt immediately guilty and petty. It was only a cooking roll. "Of course," we concluded, and reached for my glass.
Before we overwhelmed it, however, another waiter reached forward, took a potion divided and swallowed a booze in a singular gulp. "And his brother, Eric, is really thirsty," pronounced a President.
I didn't contend anything. The President is contrast my compassion, we thought. we funded my comments and motionless to play along. we don't wish to seem unkind.
My image was whisked divided before we had tasted a bite.
"Eric's children are also utterly hungry."
Then, many curiously, we crashed to a floor. My chair had been pulled out from underneath me.
I stood, brushing myself off, and watched as it was carried from a room.
"And their grandmother can't mount for long."
I immune myself, smiling outwardly, though inside feeling like a fool. Obviously we had been invited to a White House to be competition for some game. we reached for my coat, to find that it had been taken.
I incited behind to a President.
"Their grandfather doesn't like a cold."
I wanted to shout, "that was my coat!" But again, we looked during a peaceful smiling face of my horde and motionless we was being a bad sport. we widespread my hands helplessly and chuckled.
Then we felt my hip slot and satisfied my wallet was gone. we immune myself and walked to a phone on an superb side table.
After a few calls, we schooled that my credit cards had been maxed out, my bank accounts emptied, my retirement and equity portfolios had vanished, and my wife had been thrown out of a home.
It incited out that a waiters and their families were relocating into my house. The President hadn't altered or oral as we schooled all this, though finally we lowered a phone into a cradle and incited to face him.
"Andrew's whole family has finished bad financial decisions. They haven't designed for retirement and they need a house. They recently defaulted on a subprime mortgage. we told them they could have your home. They need it some-more than we do."
My hands were shaking. we felt faint. we stumbled behind to a list and faced my host.
The President cheerfully cut his meat, ate his steak, and drank his wine. we lowered my eyes and stared during a tiny grey circles on a tablecloth that were H2O drops.
"By a way," he added, "I have only sealed an Executive Order nationalizing your factories.
I'm banishment we as conduct of your business. I'll be handling a organisation now for a advantage of all mankind.
There's a whole garland of Erics and Andrews out there and they can't come to we for jobs cringing like beggars…we need to widespread YOUR resources around…"
I looked up. The President forsaken his ladle into a dull ramekin that had been his crème Brule.
He emptied a final drops of his wine. As a list was cleared, he illuminated a cigarette and leaned behind in his chair.
He stared during me. we clung to a dilemma of a list as if it were a edge and we were a male unresolved over an abyss.
I suspicion of a years behind me, of a life we had lived. The life we had warranted with a lifetime of work, risk and struggle.
Was we being punished? How had we authorised all this to happen? What diversion had we played and lost? we looked opposite a list and beheld with some warn that my horde was many gentle and relaxed.
What was a definition of all this?
As if responding a tacit thought, President Obama unexpected standing his head, sealed his dull eyes to mine, and revealed a million teeth, chuckling agreeably as he folded his hands.
"You should have stopped me during a cooking roll," he said.
So you, we standard citizen and you, we customary AMERICAN, has your cooking hurl disappeared?
Alex Jones Jewish Connections Documented
9/11 half-truther Charlie Sheen with half-ju Alex Jones
Here Alex Jones admits that his father is Jewish:
Below Jones claims a "Israeli Lobby" is swelling a story that he works for Israel. Says he "traced" a rumors to Israel. Also admits his wife "has some Jewish ancestry."
Alex Jones brags of his support for non-whites:
Alex Jones on Illegal Immigration
[Alex Jones is such an ass. Watch this video. Comes in and deliberately disrupts a pro-gun convene in Texas. Unbelievable.]
Alex Jones Wrecks Pro-Gun Rally, Then Lies About It
By Brother Nathanael Kapner, Copyright 2009
THE CONTROVERSY OF ALEX 'BULLHORN' JONES being an purported Zionist shill has now stretched to his apparent Zionist-Jewish connectors on his 1) Personal Staff 2) Website Advertisers 3) Link To Time Warner President, Edgar Bronfman Jr.
Jones' ostensible devotion to Zionism, that explains since he probably never targets Zionist Jews or a racist, Zionist brute state of Israel when enchanting in his scandalous rants and attacks, might good branch from a Protestant-Zionist faith system, and a widely circulated reports that his mother is a Jew – – that creates his twin children Jewish underneath Talmudic law and authorised for a Israeli Law Of Return. In that Jones' wife is of Jewish heritage, both she and a children would be involuntary twin citizens…Alex would not qualify.
With additional information on Jones that this site is now providing, Alex Jones' Zionist connectors call his privacy to display tellurian Zionism, becomes ever some-more obvious.
And to supplement insult to injury, Jones recently interviewed a Zionist-Jew professor, a "Dr Jonathan Levy," who railed in standard Jewish conform opposite a Catholic Church as being partial of a New World Order. Yet, not a word from Jones and his Jewish guest about Zionist Jewry being a genuine pushing force behind a NWO.
Beginning With His Personal Staff, Jones' Key Employees Are Jews:
Rob Jacobson: Jones' cameraman. Jacobson also does film and video modifying work for Alex Jones.
Aaron Dykes: He does web-mastering and video work for Jones' Websites, Infowars and Prisonplanet.
Jason Bermas: He hosts Jones' radio show, The Alex Jones Show, when Jones is on vacation. Bermas recently pounded all critics of Jews, job them, "Anti-Semites" and saying that they should all die.
Alex Jones Also Has An Abundance Of Jewish Advertisers Who 'Pay His Way:'
Henri-James Tieleman: President of EcoloBlue Life Energy. Tieleman does live, on-air 10-minute commercials on Alex Jones' radio show, and a unchanging available commercials are uttered by Alex Jones himself.
Tieleman also has a vast peep animation ensign announcement during a tip right-hand dilemma of a categorical page of Alex Jones' Prisonplanet Website.
Steve Shenk: Director of a Michael Stevens Group. Shenk advertises his subsidiary, eFoods Direct, on Alex Jones' radio show. See Shenk On The Alex Jones Show Here.
Allen L Schultz: Owner of Pre-Paid Legal Services, Inc. Schultz mostly uses an charcterised ensign announcement during a tip right-hand side of a categorical page of Alex Jones' Prisonplanet Website. View Jones' Entire Jewish Advertisers Here. [JdN: A Howard Schultz is a ultra-juish owners of Starbucks…. Even a many German-sounding name can be juish. Of course, "Allen' is a classical juish initial name.]
HOLLY BRONFMAN LEV IS Edgar Bronfman Jr's sister. As owners of a Bet Lev Foundation and 100% owners of Lev Group Ltd, Holly Bronfman's authorised affairs are conducted by a high-powered lawyer, Elizabeth Schurig, who also happens to hoop a authorised affairs of Alex Jones.
Schurig's authorised bureau is located during 100 Congress Avenue, 22nd Floor, Austin Texas 78701. Of sold note, on Jan 3, 2008, Schurig altered Jones Productions LLC to a same residence as her authorised firm, 100 Congress Avenue.
The "Governing Authority" of Jones' Infowars LLC and his Jones Report LLC are also during a residence of Schurig's law offices.
The Alex Jones Machine Website puts it all like this — "So now Alex's corporate addresses is a Bronfman's lawyer's address?" View Entire Story Here.
At a really least, a Jones revenue-streams from his Jewish sponsors benefaction distant too many obstacles for him to plead universe Zionism to any picturesque degree. Sponsor revenues aside, Jones' Jewish connectors are, indeed, profound, wide-ranging and series distant too many for him to brave news a contribution and dynamics of a Zionist control, change and confinement of many of a planet…even if he wanted to.
I am not certain that Jones is a totally bad apple. we was a Marine-trained interrogator and we watch his physique language. we consider a male IS frank though a juze have given him an ultimatum:
It is unsuitable to adore Benjamin Freedman (or for that matter, Alan Sabrosky, Israel Shamir, Jack Bernstein, etc.) and nonetheless hatred Alex Jones. Jones has finished a lot of good. But he is a half-ju and married to a ju-ess. That fact will perpetually put us on a guard.
We contingency learn from all a enemies to be MORE SOPHISTICATED — and one of those enemies, who was brilliant, was Leon Trotsky. Trotsky pronounced that some organizations, by being "moderate," acted as "conveyors belts."
That is, they move people along from a Establishment position toward the some-more radical and also some-more guileless position.
So we need an Alex Jones and we need a Ron Paul. We even need (clothespin on nose here!) FOX and Limbaugh, nonetheless they are totally controlled. Yet they really effectively conflict Obama, a black poser messiah, who could have enacted hatred debate laws and martial law by now following another false-flag conflict — if FOX had not been around. FOX is a number-one news network!
But it is also time for THE REAL MCCOY. The loyal jufighter. To contend a juze did 9/11, protocol murder, their association Verint controls a phones, their association Skype reads a IMs, that WWI and II were crimes opposite peace, opposite amiability and fight crimes, that Dresden was an atrocity, that a Israelis are not even biblical juze though Khazars, and that for all this faults, on a categorical things HITLER WAS RIGHT.
Especially about a pleasing prophesy and unsentimental judgment of rebuilding a naoitn around a "folk community": we are nothing; your folk is everything.
The time is now for National Socialism to lapse in an AMERICAN form.
That is Solutrean amicable nationism. Stay tuned and, to be blunt, greatfully DO send money. If we are feeling doomed, do something constructive with that tension and assistance with money, or proffer time, mechanism skills, or only promulgation links to my website to friends. And call me! (My series is on a Contact Page above, right where it should be.) we only fielded a call currently from a lady comrade who woke adult after being on a ju-infested chosen college campus. It was in college that we too awakened.
I am really receptive and never too bustling to speak with another white hermit and sister. The word "Namaste" is not an dull word for me. we do worship a God within you.
Even within dogs….
My work could have begun months ago with your financial support. we had a family male prepared to come here and assistance me full-time and reinstate Henrik Holappa. But though supports we could not means him. But do we know ANYONE else with a earthy courage, prophesy and a totally new concept? Determined not to offer a universe some-more of a — accurate though ungalvanizing — white jingoist same-old, same-old? The "convert-people-one-at-a-time" method?
What we need now is a MASS movement.
I now have FRENCH supporters….where Jean-Marie Le Pen's party, a Front National, has only finished an extraordinary comeback, removing 9 percent of a opinion national and scoring 23% in a south!
I have countless German supporters, in Deutschland, Austria and elsewhere around a world….
and in Britain, home of many of my ancestors…..
But THEY ALL KNOW ………
that ONLY AMERICA…………..
the land of Constitutionally-guaranteed giveaway debate (which can't even be "abridged" even if it is called "hateful"), giveaway debate buttressed by gun rights, can save a white world.
Link to this story:


text:
Skip to content
and found 2 items
625cc, 54bhp, 110mph, Insurance group 11
If you're used to smooth riding, civilized singles like BMW's F650 range then KTM's 640 Adventure will come as something of a shock. It's very vibey, absurdly tall, not that comfortable and pre-2004 models suffer from patchy engine reliability. Having said that in the muddy side of its dual-purpose role it's untouchable for handling, presence and rugged toughness.
904cc, 68bhp, 128mph, Insurance group 11
On face value the Cagiva Elefant was born of an unlikely marriage, but the reality of mating up the (then Cagiva owned) Ducati V-twin with a monster trailie, desert racer type chassis proved something of a masterstroke let down only by poor build quality. The big Elefant (a 750 was also produced) was lively, handled fairly well and was even ...
Save money by comparing quotes. It's quick and easy
With over 18,000 products, MCN shop is the best place to buy your biking kit
Motorcycle Cover | Cheap Motorcycle Insurance | Moped Insurance | Scooter Insurance | 125 Insurance | Work Experience | Complaints Policy
Car Magazine | Empire | FHM | Parkers | LRO | Today's Golfer | Zoo
Bauer Consumer Media Ltd (company number 01176085, 1 Lincoln Court, Lincoln Road, Peterborough, England PE1 2RF)