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Dan
Daniel L Fouts
Over $ 1.3 trillion a year spent on
U.S. HEALTH CARE.
That is more per person than any country
in the world!
That 1.3 Trillion Dollars bought us
# 37
In the world in ability to achieve vital
health goals according to the World
Health Organization’s
World Health Report 2000
FOU R M AJOR
CHA LLE NGES IN
HEA LTH C ARE TOD AY
Chronic degenerative
diseases -120+ million
Americans
Autoimmune diseases -
50+ million Americans
Infectious diseases -
multiple anti-biotic
resistant strains
CHILDR EN A RE DEVELOP ING
“A DULT ONS ET” DI SEASE S AT
A FRIGHTE NING RATE
Hundreds of medical studies in the last
few years are looking at viruses and
bacteria as etiological factors in diabetes,
heart disease and heart attacks.
The closing years of the second millennium have been uplifting for
carbohydrate biology. Optimism that oligosaccharide sequences are bearers
of crucial biological information has been borne out by the constellation of
efforts of carbohydrate chemists, biochemists, immunochemists, and cell-
The direct involvement of specific
and molecular biologists.
oligosaccharide sequences in protein targeting and
folding, and in mechanisms of infection,
inflammation and immunity is now unquestioned.
With the emergence of families of proteins with carbohydrate-binding
activities, assignments of information content for defined oligosaccharide
Dr. Mondoa’s book
is helping to
change the old
biochemical
paradigm that
carbohydrates are
just a fuel supply
for the body. The
tremendous
potential of this
special group of
bioactive sugars
promises to change
AnnuRevBiochem 1976; 45:217-37
Comparativeaspects ofglycoproteinstructure.
Kornfeld R,KornfeldS.
Glycoproteinshaveawidedistributioninnatureandserveavastnumber of functions.
Thereareglycoproteinenzymes andhormones;glycoproteinsare foundinbloodand
secretions, in cell membranes, and in connective tissue. Of all the biologically
occurringmacromolecules theglycoproteins, which consistof carbohydratemoieties
convalently linked to a polypeptide backbone, represent themost diverse group,
rangingfromsubstances inwhichthecarbohydrate component represents less than
1% of the total weight to those in which It represents over 80% of thetotal. The
proteoglycans, which areclassified separately from otherglycoproteinsandinclude
thechondroitinsulfates,dermatansulfates, andheparinprimarilycarbohydrateinthe
form of numerous heteropolysaccharide chains attachedto apolypeptidechainat
closely spacedintervals. Thesugars thatcommonlyoccur inglycoproteins
include galactose, mannose, glucose, N-acetylglucosamine, N-acetyl-
galactosamine, sialicacids, fucose, and xylose. The proteoglycans also
containvariousuronicandsulfatedamino sugars.
• Structural molecules • Immunolgic
- cell walls
molecules -
immunoglobulins
- collagen, elastin
-
-
histocompatability antigens
fibrins
- bone • Enzymes
matrix
- proteases
• Lubricants and
protective agent - nucleases
-
-
glycosidases
mucins
- clotting
factors
- mucous secretions
• Cell attachment /
• Transport molecules recognition site
for - vitamins
Science 2001 Mar 23;291(5512):2370-6
Lectins, non-enzymic proteins that bind mono- and oligosaccharides reversibly and
with high specificity, occur widely in nature. They come in a variety of sizes and
shapes, but can be grouped in families with similar structural features. The
combining sites of lectins are also diverse, although they are similar in the same
family.The specificities of lectins are determined by the exact shape of the binding
sites and the nature of the amino acid residues to which the carbohydrate is
linked. Small changes in the structure of the sites, such as the substitution of only
one or two amino acids, may result in marked changes in specificity. The
carbohydrate is linked to the protein mainly through hydrogen bonds, with added
contributions from van der Waals contacts and hydrophobic interactions.
Coordination with metal ions may occasionally play a role too. Microbial surface
lectins serve as a means of adhesion to host cells of viruses (e.g. influenza virus),
bacteria (e.g. E. coli) and protozoa (e.g. amoeba): a prerequisite for the initiation
Blocking the adhesion by carbohydrates that
of infection.
mimic those to which the lectins bind prevents infection
by these organisms. The way is thus open for the
development of anti-adhesive therapy against microbial
diseases. Lectin-carbohydrate mediated interactions
between leucocytes and endothelial cells are the first
the most attractive strategy for developing
anti-adhesive thera-peutic agents is to use
soluble forms of the human oligosaccharide
component, which are small and non-
“A striking example of the successful
immunogenic.”
application of antiadhesive, soluble
carbohydrates involved application of a
mixture of sialyated complex N-linked
glycopeptides (500 mg/day) to protect
newborn calvesforfrom
Drug prospects a oligosaccharides
use of lethal dose of
enterotoxic E coli
include otitis 99.” pneumonia, chronic
media,
bronchitis, gastrointestinal diseases, cystitis,
gonorrhea, chlamydia Lancet
and1996;possibly
347:
enhancement of the effects of standardized
1017-21
Eur J ClinMicrobiol 1987Oct;6(5):591-3
In vitro experiments with frozen sections of human lung and kidney demonstrated that
adhesion of Streptococcus pneumoniae Pn 629 Type 14 and Pseudomonas
aeruginosaATCC 27853 to human cells was mediated by bacterial lectins (adhesins)
with N-acetyl-D-glucosamine/D-galactose or N-acetyl-neuraminic acid specificity.
Blockingof thelectinbindingsites onbacterial surfaces withcompetitive
carbohydrates completely prevented the bacterial adherence, whereas
non-specific carbohydrates (D-mannose, D-xylose) did not inhibit
adherence.
BACTERIA BACTERIA(cont’ VIRUSES
Actinomyces d) Proteus HIV
naeslundil mirabilis
Bordetella Pseudomonas Influenza virus
pertussis aeruginosa Parvovirus
Chlamydia Burkholderia Rotavirus
pneumoniae cepacia Salmonella
Citrobacter freundi typhimurium
--------------------------
Enterobacter Serratia
aerogenes ------------
marcescens
Escherechia coli Shigella
Haemophilius dysenteriae FUNGI
influenzae H S flexneri Candida
parainfluenza Staphylococcus albicans
Helicobacter pylori aureus S
Klebsiella saprophyticus --------------------------
pneumoniae Streptococcus
Specific oligosaccharides shown to be effective
------------
Mycobacterium
antiadhesive agents inmutans
vitro. S (Lancet,1996;
tuberculosis
Specific oligosaccharides
pneumoniae
shown to
OTHER
be prophylactic347:1017)
or
ANTI-INFECTIVE AGENTS
Adhesion of bacteria and of metastasizing tumour cells have much in common, especially the
participation of lectins in this process. In the future it might be possible to inhibit the
metastatic process and bacterial adhesion by blocking with lectins specific
for appropriate (oligo) saccharides or glycoconjugates. Initial clinical trials
arevery promising.
Cancer cells have unusual carbohydrates on
their surface, which may account for many of
their invasive properties. Drugs that interfere
with the adhesiveness of abnormal cells may
someday be used in cancer therapies.
Sharon and Lis, “Carbohydrates in Cell Recognition,” Scientific American, January 1993, pgs. 82-
89
• Billions of dollar$ are being
spent to create synthetic
carbohydrate drugs because the
effectiveness of natural
carbohydrates have been
demonstrated repeatedly in
medical research.
• GLYCONUTRIENTS are
available for patients NOW -
Anticancer Res 1997 Mar-Apr;17 (2B):1223-6
Prevention of hepatic
metastases by liver lectin block-ing
with D-galactose in stomach cancer
patients. A prospectively randomized
clinical trial.
Kosik J, Gil J, Szmigielski S, Beuth Jm Pulverer G
Postgraduate Medical School, Warsaw, Poland
TheEffectof L-FucoseonRatMammaryTumor
Growth. II. InVitro Studies
J ames M. Roseman, A.B., Elizabeth Miller, Ph.D., Murray H. Seltzer, M.D., Daniel Wolfe, B.S., and Francis E
Rosato, M.D.
Different concentrations of L-fucose uniformly produced a suppression in the growth rate and a change in the
morphology of cells grown in tissue culture. The inhibition of growth of these malignant cells
was found to be concentration dependent with 100% inhibition of growth at a
concentration of 50 mg fucose per milliliter of medium and 60% inhibition at a
concentration of 12.5 mg per milliliter medium. Using other sugars that are
components of glycoproteins, it was shownthatmannoseandgalactosecouldalso
depress the rate of growth of these tumor cells in culture. When 0.05 ml of packed tumor
cells used in these experiments was resuspended in 1 ml of mediumand injected into a rat, a tumor grew at
the site of injection. This newtumor exhibited similar growth characteristics and showed the same histological
appearance as the tumor fromwhich the cell line was derived.
Acta Univ Palacki Olomuc Fac Med 1989;122:113-20
syndrome
type I.
Carbohydrate-deficient glycoprotein syndrome type I
(CDGS) is an inherited metabolic disorder with
multisystemic abnormalities resulting from a failure to add
entire N-linked oligosaccharide chains to many
glycoproteins. Fibroblasts from these patients also abnormally glycosylate
proteins, but this lesion is corrected by providing 250 microM mannose to the
culture medium. This correction of protein glycosylation suggests that providing
dietary mannose to elevate blood mannose concentrations might also remedy
some of the underglycosylation observed in these patients. We find that ingested
mannose is efficiently absorbed and increases blood mannose levels in both
normal subjects and CDGS patients. Blood mannose levels increased in a dose-
dependent fashion with increasing oral doses of mannose (0.07-0.21 g
mannose/kg body weight). Peak blood mannose concentrations occurred at 1-2 h
following ingestion and the clearance half-time was approximately 4 h. Doses of
0.1 g mannose/ kg body weight given at 3-h intervals maintained blood mannose
conentrations at levels 3- to 5-fold higher than the basal level in both normal
J ClinInvest1996Mar 15;97(6):1478-87
Mannosecorrects alteredN-glycosylationin
carbohydrate-deficientglycoproteinsyndrome
fibroblasts.
PanneerselvamK, FreezeHH; LaJollaCancer ResearchFoundation,
Type I carbohydrate-deficient glycoprotein syndrome (CDGS) patients fail to add entire N-linked oligosaccha-
ride chains to some serumglycoproteins. Here we showthat four CDGS fibroblast cell lines have two related
glycosylation abnormalities. First, they incorporate 3-10-fold less [3H] mannose into proteins, and, second,
the size of the lipid-linked oligosaccharide precursor (LLO) is much smaller than in controls. Addition of
exogen-ous mannose, but not glucose, to these CDGS cells corrects both the lowered [3H] mannose
incorporation and the size of LLO. These corrections are not permanent, and the defects immediately
reappear when mannose is removed. To explore further the basis of mannose correction, we analyzed the
amount of 3H- labeled LLO intermediates. Except for dolichol-P-mannose, other precursors, including
mannose, mannose-6-phosphate, mannose-1-phosphate, and GDP-mannose, all showed a 3-10-fold
decrease in CDGS cells. Thus, there are no obvious lesions in the intracellular conversion of mannose into
LLO, and, once inside the cell, [3H]mannose appeared to be metabolized normally. Initial velocities of
[3H]mannose uptake were two-to threefold less in CDGS cells compared with controls, and this slower
transport may partially explain the re-duced [3H]mannose incorporation in CDGS cells. Since we previously
showed that the enzymes converting glucose to mannose-6-phosphate appear to be normal, our results
suggest that cells may acquire or generate mannose in other ways. Although we have not identified the
primary defect in CDGS, these studies showthat intracellular mannose islimited
and that some patients might benefit from including mannose in their
regular diets.
J ClinInvest1998Apr1;101(7):1414-20
Kornfeld S.
Department of Internal Medicine, Washington University School of Medicine, St.
Louis, Missouri 63110.