Natural Remedies for Food Cravings

Sugar Crush Detox

This program was designed by Jane who had the same problems with sugar. Throughout her life, she was addicted to sugar and she thought she needs swift intervention before that habit develops into something else. She had an experience that helped her beat sugar addiction with the rest of the world. Her program helps you cut all the roots of majority of the health problems you usually gets. It attacks the weight loss problem at its source which is the biological craving for sugar. This product was specifically created to help people with sugar cravings beat this addiction and lead a healthy life. This program contains a couple of guides available in PDF, MP3 and video formats. The author used simple language in all the formats to ensure that everybody will be able to handle sugar addiction. If you are one of them and you want to get the full support required to quit sugar and lead a heathy life, then Sugar Crush Detox is for you. Read more...

Sugar Crush Detox Summary

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Maltose Sucrose Lactose Are Important Disaccharides

The physiologically important disaccharides are maltose, sucrose, and lactose (Table 13-4 Figure 13-11). Hydrolysis of sucrose yields a mixture of glucose and Sucrose fructose which is called invert sugar because the strongly levorotatory fructose changes (inverts) the previous dextrorotatory action of sucrose.

Fantastic tliar teeding formula

Foliar (or leaf) feeding is the most efficient way to fertilize, says John Dromgoole, owner of Garden-Ville nursery in Austin, Texas, and host of the Gardening Naturally radio program. When you apply fertilizer to the soil, the roots may take up as little as 10 percent of the nutrients, he explains, but when fertilizer is applied to the leaves, 90 percent of the material is absorbed. John regularly foliar-feeds all of the nursery stock at Garden-Ville, as well as the plants in his home landscape. His formula includes fish emulsion for nitrogen and seaweed for trace minerals, growth stimulants, and plant hormones. One of John's secret ingredients is blackstrap molasses, which contains iron and sulfur as well as simple sugars that nourish the plants.

Background And Relevant Pharmacokinetics

Calcium's bioavailability from both food and supplements shows enormous variation, from 4 to 45 (Recker 1985) and is dramatically influenced by other foods present in the gastrointestinal tract. Phytates, oxalates, all types of fibres, unabsorbed dietary fatty acids and other divalent minerals all potentially compromise its absorption, while lactose (especially in children) and other sugars, as well as protein and the presence of vitamin D all enhance uptake (Groff & Groper 2000).

Paroxysmal Nocturnal Hemoglobinuria

Paroxysmal nocturnal hemoglobinuria (PNH) is another clonal hematopoietic disorder to consider in the differential diagnosis. It is characterized by a defect in the glycosylphosphatidylinositol (GPI)-anchor due to mutations in the PIG-A gene (Rosse 1997). The loss of many proteins bound to the GPI-anchor on the surface of hematopoietic cells is thought to lead to the clinical manifestations of hemolysis, venous thrombosis, and bone marrow failure. Furthermore, given its clonal nature, it has also been described in the setting of MDS, myeloproliferative disorders, and the progression to AML (Longo et al. 1994 Nakahata et al. 1993). The diagnosis of PNH should be considered in any patient presenting with cytopenias and a hypocellular bone marrow. Classically diagnosed by the sucrose lysis and Ham's test, it may now be identified by the absence of GPI-linked proteins, CD55 and CD59, on the surface of peripheral blood cells by using monoclonal antibodies and flow cytometry. Flow...

C1anion Approaches

The chemistry associated with the anomeric center is cationic by nature. One interesting approach to C-glycoside synthesis is to reverse the electronic character of this center from electrophilic to nucleophilic. Hence anions at the anomeric center (gly-cosyl anions) have now become fairly common in the synthesis of C-glycosides. Of course, one limitation of this method was the presence of heteroatoms at C2, since any discrete anion at C1 would surely cause an elimination reaction to occur and in turn produce the corresponding glycal (55 58, Scheme 13). This is in fact a very common way of preparing sugar glycals from sugars. As a consequence of this limitation, the chemistry of 2-deoxy C1 anions (56 59) has been extensively explored. An interesting development in this area came with the observation that if the hydroxyl group at O2 is unprotected, deprotonation followed by anion formation at C1 can occur to give species 57, which can go on to react with a suitable elec-trophile to...

Secondary Deficiency

Factors that may exacerbate deficiency, generally by increasing requirements for or urinary excretion of chromium, include pregnancy, excessive exercise, infection, physical trauma and stress (Anderson 1986). Diets high in simple sugars have been found to increase urinary chromium excretion up to 30-fold, thereby increasing the risk of deficiency (Kozlovsky et al 1986). Corticosteroids also increase urinary losses of chromium (Kim etal 2002).

Detailed Preparation of 500 ml

Dissolve 3.03 g Tris (Boehringer Mannheim) and 85.575 sucrose (BBL) in 250 ml H20. Adjust pH to 8.4 with 5 N HC1. Bring to 300 ml. Autoclave for 15 min in a 500-ml bottle. 4. To the cooled 300-ml Tris sucrose solution, add 25 ml of the following sterile stocks 10 NaCl (filter), 10 yeast extract (autoclave), 10 Bacto-Tryptone (autoclave), and 10 glucose (filter). Then add 1 ml of these filter-sterilized stocks 0.5 M CaCl2 (3.68 g of CaClr2H20 in 50 ml H20) , 1.0 M MgS04 (12.32 g of MgS04-7H20 in 50 ml H2OJ, and 0.5 M MnS04 (1.69 g of MnS04 H20 in 50 ml H20).

Psychological Effects

Chocolate craving, which is reported to be the most common food craving (Weingarten & Elston 1991), is more common in women, with fluctuations occurring with hormonal changes just before and during the menses (Rozin et al 1991). The basis for chocolate craving, however, remains undetermined, but it is suggested that aroma, sweetness, texture and calorie content are likely to play a more important role 2007 Elsevier Australia in chocolate cravings than pharmacological factors (Bruinsma & Taren 1999, Michener & Rozin 1994, Rozin et al 1991, Smit et al 2004).

Examination screening specific to children

Note Formal audiological evaluation should be carried out at any time if there is clinical suspicion or parental concern. No simple screening test is very reliable for sensorineural or conductive deafness. Testes. Screen at birth, and 6-8 weeks, 6-9 months and 3 years for absence or maldescent. Those who have been treated for maldescent have a higher risk of neoplastic development in adolescence. Dental assessment fluoride. Advise daily fluoride drops or tablets, if water supply is not fluoridated. Children's teeth should be checked regularly, particularly if a school dental service is not available. Advice should be given on sugar consumption, especially night-time bottles, and tooth cleaning with fluoride toothpaste to prevent plaque.

Premenstrual Syndrome

Magnesium deficiency may contribute to the symptomology of PMS, which may be improved by chocolate or cocoa powder, which contain a high concentration of magnesium ( 100 mg 100 g in chocolate and 520 mg 100 g in cocoa powder). There is also some evidence to suggest that serotonin levels are low premenstrually, and it is possible that premenstrual chocolate cravings are the body's attempt to raise CNS concentrations of serotonin (Bruinsma & Taren 1999).

Conformational Studies

A detailed appreciation of the biological function of a molecule requires that its conformation be understood. Therefore, in recent years a great deal of effort has been directed toward elucidating the solution conformation of oligosaccharides 92 . The work, however, has been heavily focused on oligopyranosides. At the same time, a wealth of information detailing the conformation of furanose rings in nucleotides and nucleic acids has accumulated 93 . However, except for sucrose 94,95 , only recently have conformational investigations of oligosaccharides containing furanose residues been reported 96,97 . Outlined in this section are some recent investigations that have explored the solution conformation of methyl a-d-arabinofuranoside and oligosaccharides containing a-d-arabinofuranosyl residues.

[1 Arrays of Transfected Mammalian Cells for High Content Screening Microscopy

Add 7.5 ul EC buffer (EC Buffer is part of the Effectene Transfection kit, Qiagen) containing 0.2 M sucrose and mix thoroughly by pipetting three times up and down. It is important to mix the transfection components just prior to the spotting, to achieve a high reproducibility in transfection efficiency. The optimal incubation times and amounts of transfection reagent are determined empirically for different transfection reagents. However, for optimizing the transfection mix for transfection reagents different from Effectene, the protocol described previously is a good starting point for optimization by simply replacing Effectene with equal amounts of the alternative transfection reagent. The EC buffer from the Effectene kit can be replaced by water without significant loss of transfection efficiencies, when tranfection reagents different from Effectene are used. The presence of sucrose in the EC buffer reduces the loss in transfection efficiencies when the dried arrays are stored...

Mass Spectra Interpretation

A systematic nomenclature for labeling fragment ions observed in MS MS spectra has been introduced by Domon and Costello in 1988 (12). As shown in Scheme 1, Ai, B , and C are used to designate fragments containing a terminal (nonreducing end) sugar unit, whereas Xj, Yj, and Zj represent ions still containing the aglycone or the reducing sugar unit. Subscripts indicate the position

Two Basic Strategies

The chemical synthesis of oligosaccharides has been a major focus since the 1980s 8 as a result of our increased understanding of the importance of these sugars in many life processes 3 . Although with low efficiency and sometimes with tremendous difficulties, almost all the sugar linkages known in nature have been synthesized.

Generation of Adenoviruses by Manipulation of Viral DNA

Recombinant adenoviruses were originally created by isolating fragments of the viral chromosome, followed by ligation or recombination to regenerate a complete viral backbone. Generally, DNA isolated from virions is digested with a restriction enzyme that removes the left end of the chromosome, including the viral packaging signals. This renders the large DNA fragment noninfectious. The unique Clal site located at 2.6 m.u. is commonly used for this purpose (Fig. 3A). Undigested viral DNA and the smaller DNA fragments are often removed by sucrose gradient ultracentrifugation. The large fragment is ligated in vitro to a DNA fragment containing the left end of the chromosome and containing the desired modifications (Stow, 1981). If the viral DNA contained in this fragment has a deletion of the Ela region, the resulting recombinant Ad will be a replication-defective El mutant. Inclusion of unique restriction sites in the left end-containing plasmid (generally in the place of the Ela gene)...

Primary treatment strategies

Symptoms related to fluid retention can usually be eradicated through dietary measures, most specifically restriction of sodium and simple sugars. However, diuretics may be useful in patients with very troubling edema. Spironolactone has bee demonstrated to be effective in a dosage of 100 mg per day.

Plant Part Used Chemical Components

Dong quai contains essential oil (0.4-0.7 ) consisting of 45 ligustilide, n-butylphthalide, cadinene, carvacrol, safrole and isosafrol. The root also contains sucrose (40 ) and various lactones and vitamins, together with phytosterols, ferulic acid and coumarins, including osthole, psoralen and bergapten (Micromedex 2003). Ferulic acid and ligustilide are considered to be the main active components (Dong et al 2005) and it has been suggested that assessment of total ferulic acid content provides a good measure of herbal quality (Lu et al 2005).

Carbohydrates and mental health

Carbohydrates include starches, naturally occurring and refined sugars, and dietary fiber. Foods rich in starches and dietary fiber include grain products like breads, rice, pasta and cereals, especially whole-grain products fruits and vegetables, especially starchy vegetables like potatoes. Foods rich in refined sugars include cakes, cookies, desserts, candy, and soft drinks. Some researchers claim that a high sugar intake causes hyperactivity in children. Although carefully controlled studies do not support this conclusion, high sugar intake is associated with dental problems. Further, foods high in refined sugars are often low in other nutrients, making it prudent to limit their use.

Biomedical Importance

Carbohydrates are widely distributed in plants and animals they have important structural and metabolic roles. In plants, glucose is synthesized from carbon dioxide and water by photosynthesis and stored as starch or used to synthesize cellulose of the plant framework. Animals can synthesize carbohydrate from lipid glycerol and amino acids, but most animal carbohydrate is derived ultimately from plants. Glucose is the most important carbohydrate most dietary carbohydrate is absorbed into the bloodstream as glucose, and other sugars are converted into glucose in the liver. Glucose is the major metabolic fuel of mammals (except ruminants) and a universal fuel of the fetus. It is the precursor for synthesis of all the other carbohydrates in the body, including glycogen for storage ri-bose and deoxyribose in nucleic acids and galactose in lactose of milk, in glycolipids, and in combination with protein in glycoproteins and proteoglycans. Diseases associated with carbohydrate metabolism...

Alternative Substrates

While isozymes are apt to have different energies of activation, even under the same assay conditions, an enzyme acting on different substrates can in some circumstances exhibit the same energy of activa-tion10,11. Yeast sucrase, for example, has an energy of activation of 46 kJ-mol 1 (or 11.0 kcal-mol-1) for both sucrose and raffinose11-13. The rate-determining step in the enzyme-catalyzed reaction may differ with alternative substrates, and this may be reflected in the observed energy of activation. Likewise, if the rate-determining step changes with protein modification, assay conditions, or through site-directed mutagenesis, Arrhenius plots should reflect those changes. An example is the myosin ATPase which exhibits biphasicity in the Arrhenius plot with ITP as a substrate, but a typical linear Arrhenius plot with ATP as the substrate. Levy, Sharon, and Kosh-land14 suggested that this may be the result of the 6-amino group on ATP interacting with some functional moiety...

Chromatin Remodeling Enzymes

The ability to remodel nucleosomes is vital for accurate and regulated transcription and efficient DNA replication. Nucleosome remodeling is defined as an enzymatic activity capable of altering the position or stability of the nucleosome. The flagship of the ATP-dependent, enzyme complexes that remodel chromatin is the SWI SNF complex. The SWI SNF class of chromatin remodeling complexes is named for the genetic screens used to identify sucrose non-fermenting (Snf) or mating-type switching defective (Swi) mutants. The SWI SNF chromatin remodeling complexes contain an enzymatic ATPase protein subunit, which on its own is capable of remodeling chromatin in vitro (Cote et al., 1994). In yeast this protein is called Snf2 Swi2 and in humans there are two homologous proteins Brgl and Brm. Each of these proteins contains a structural motif called a bromodomain, which binds to acetylated lysines. Acetylation of histones by HATs promotes recruitment of SWI SNF remodeling complexes through...

Other Cytokine Receptor Superfamily Members

The notion of preformed dimers on the cell surface is not solely associated with the cytokine receptor super-family. The epidermal growth factor (EGF) plays important roles throughout development including cell proliferation, differentiation, and survival of multicellular organisms 29 . The EGF receptor (EGFR) is a member of the growth factor receptor tyrosine kinase family. Using chemical cross-linking experiments and sucrose density-gradient centrifugation, experiments have demonstrated that EGFR forms a dimer in the absence of bound ligand 30 . A flexible rotation model was proposed for EGFR activation in which the binding of EGF induces rotation of the juxtamembrane domain and, accordingly, the transmembrane domain. Consequently, the dimeric intracellular domains dissociate, allowing for the catalytic kinase domains to become accessible to their substrate tyrosine residues.

Production of Secondary Metabolites by in vitro Culture

Furthermore, it was found that semicarbazide and particularly thiosemicarbazide inhibited the production of phytosterols and promoted the biosynthesis of saponins in the presence of mevalonic acid (Furuya et al, 1983b, Linsefors et al., 1989). Odnevall and Bjork (1989a) who studied the relationship between the morphological state and ginsenoside formation found that the maximum ginsenoside production occurred in tissue cultures consisting of cell aggregates and differentiated roots on a medium supplemented with 2,4-D and kinetin under light conditions after 2-4 days in the stationary phase. The dynamics of the biosynthesis of ginsenosides during a one growth cycle of callus cell culture of ginseng was studied by Konstantinova et al. (1995) who found that maximum ginsenoside accumulation occurred on the 50th-80th day of subculture. Ginsenoside accumulation in ginseng root cultures depended on the carbon source (Odnevall and Bjrk, 1989b). Maximum ginsenoside content...

Heptadeca1 en46diyn39diol R H 513 Panaxytriol R OH

Not surprisingly ginseng roots yield about 5 per cent by weight of sugars which include the monosaccharides D-glucose, D-fructose and D-rhamnose, the disaccharides sucrose and maltose, and trisaccharides such as a-maltosyl-jS-D-fructofuranoside, (1 2)- -D-fructofuranoside and The principal sugar in fresh white ginseng root is sucrose, forming 92-94 per cent in 2 year old roots but decreasing in older roots (Sohn et al., 1988). In red ginseng root the main sugars are sucrose and rhamnose.

DNAs Plasmids Cloning and Transfection

30 Acrylamide to-acrylamide solution (29 1 Sigma), N,N,N,N'-tetramethyl-ethylenediamine (TEMED Biorad, Hercules, CA,), and 66 (w v) sucrose. Store at 4 C. 5. 5X Laemmli (5) SDS-sample buffer 0.5 M of Tris-HCl (pH 6.8), 7.5 (w v) SDS, 25 mM of EDTA, 5 M of sucrose, and 0.05 (w v) of bromophenol blue. Store at -20 C.

Cyclopropane ring fission

Both enzyme quantity and total enzyme activity. This is caused by an increase in the half-life of the enzyme from 2.3 to 3.9 h. A2059 . In contrast, treatment with sucrose or ethanol for two days resulted in no change in the amount of 'holoen-zyme', but a 30 per cent decrease in 'total enzyme'. The effect of cortisol in conjunction with sucrose appears to result entirely from decreased 'holoenzyme', but with no effect on 'total enzyme'. On its own, cortisol markedly increases both enzyme forms and eliminates the effects of ethanol. In this study it appears that the two forms were distinguished by measuring activity with and without added haematin A2062 . Rat cerebral activity is increased by administration of l-tryptophan A1289 .

Western Immunoblotting of Intact C2GnTl and the EGFPConjugated Protein

Prepare 3-mm-thick SDS gel by mixing 2.6 mL of 30 acrylamide Ms-acrylamide, 1.4 mL of separating buffer, 1.2 mL of 66 sucrose, and 5.1 mL of water. Add 104 pL of 10 SDS, 60 pL of 10 ammonium persulfate, and 6 pL of TEMED. Pour the gel (leaving space for a stacking gel) and overlay with water. The gel should polymerize in 20-30 min. 3. Prepare stacking gel by mixing 700 pL of 30 acrylamide to-acrylamide, 700 pL of stacking buffer, 600 pL of 66 sucrose, and 3.2 mL of water. Add 52 pL of 10 SDS, 30 pL of ammonium persulfate, and 3 pL of TEMED. The gel should polymerize within 30 min.

The Liver Plays A Central Role In Lipid Transport Metabolism

The fed state rather than the starved state (2) the feeding of diets high in carbohydrate (particularly if they contain sucrose or fructose), leading to high rates of li-pogenesis and esterification of fatty acids (3) high levels of circulating free fatty acids (4) ingestion of ethanol and (5) the presence of high concentrations of insulin and low concentrations of glucagon, which enhance fatty acid synthesis and esterification and inhibit their oxidation (Figure 25-6).

Sweet Taste Suppression

The constituent, gymnemic acid, inhibits the ability to taste sweetness in animal models (Fushiki et al 1992, Harada & Kasahara 2000, Kurihara 1969, 1992) and humans (Frank et al 1992). In humans, the administration of 5 mmol L gurmarin to the tongue raised the threshold ability to taste sucrose from 0.01 mol L to 1 mol L for several hours. It is suggested that gurmarin acts on the apical side of the taste cell, possibly by binding to the sweet taste receptor protein (Miyasaka & Imoto 1995).

Sugars and carbohydrates

Carbohydrates are found in simple sugars, as constituents of glycoproteins and glycolipids, and in polymeric form as structural features of bacterial and fungal cell walls, among other sources. Polymeric sugars include dextran, used as a blood expander, and starch. Sugars are most commonly analyzed by ion-exchange or other chromatographies, since they tend to be so hydro-philic as to elute rapidly from reversed phase columns. N-butyldeoxynojir-imycin, deoxynojirimycin, and their degradation products were separated on a Zorbax C8 Rx column in water acetonitrile containing heptane sulfonate and acetic acid.88 The amino sugars mannosamine, galactosamine, and glucosamine extracted from tobacco were derivatized with o-phthalde-hyde and analyzed in THF (tetrahydro furan) water on C18.89 Another deriva-tization agent, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, available commercially as AccQ-Tag , has been used for amino sugars such as a- and P-galactosamine and a- and P-glucosamine in...

Chemical Components

Other important constituents include flavonoids (isoflavonoids, liquiritin, isoliquiritin, liquiritigenin, formononetin, glabridin and the chalcones isoliquiritigenin, licochalcone A and B ) sterols (beta-sitosterol) polysaccharides (arabinogalactans) coumarins (glycerin) glabrol amines glucose, sucrose resin and volatile oil (Blumenthal et al 2000).

Separation Of Dna In The Ultracentrifuge

Velocity or equilibrium ultracentrifugation of DNA represents the only serious alternative to DNA electrophoresis. For certain applications it is a powerful tool. However, for most applications, the resolution of ultracentrifugation just isn't high enough to compete with electrophoresis. DNA can be separated by size in the ultracentrifuge by zonal sedimentation. Commonly density gradients of small molecules like sucrose are employed to prevent convection caused by gravitational instabilities. Sucrose gradient sedimentation is tedious because ultracentrifuges typically allow only half a dozen samples to be analyzed simultaneously.

Paroxysmal Nocturnal Hemoglobinuria PNH

The diagnosis of PNH should be suspected in patients with pancytopenia and thrombosis. The classic nocturnal hemoglobinuria is a rare finding. Most patients will have pancytopenia although rare patients can present with elevated blood counts. Patients will usually have a high serum LDH. The older Hams test and sucrose hemolysis tests have been replaced by flow cytometry. Flow cytometry will directly detect membrane linking proteins. The link protein CD59 is assayed and PNH is diagnosed if more than 5 of the cells are missing this protein. This technique is very sensitive to detecting small populations of cells missing proteins.

Historical Information

Boron is thought to have a direct effect on sugar synthesis. In cowpeas (Vigna unguiculata Walp), acute boron deficiency conditions increased reducing and nonreducing sugar concentrations but decreased starch phosphorylase activity (21). Under boron deficiency, the pentose phosphate shunt comes into operation to produce phenolic substances (22). Boron-deficient sunflower seeds showed marked decrease in nonreducing sugars and starch concentrations, whereas the reducing sugars accumulated in the leaves (23). This finding indicates a specific role of boron in the production and deposition of reserves in sunflower seeds. High concentrations of nonreducing sugars were also found in boron-deficient mustard (Brassica nigra Koch) (24). Camacho and Gonzalas (19) also found higher starch concentration in boron-deficient tobacco plants. In low-boron sunflower leaves, starch decreased, but there was an increase in sugars and protein and nonprotein nitrogen fractions (25). In

Diffusion vs Convection

The effect of different sizes of molecules and their distribution within brain tissue was studied by several investigators (11,12). Bobo et al. measured the volume of distribution of small (14C-sucrose, Mr 359) and large molecules (111In-Transferrin, Mr 80,000) (10). After initiating interstitial infusion, the flow rate was gradually increased from 0.5 L min to 4 L min and maintained at the higher level for 24 h. The VD within brain tissue increased linearly with the volume of infusion (VI) of both molecules, although 14C-sucrose distributed faster and occupied a larger volume compared with 111In-Transferrin. Importantly, there was no evidence of any increase of intracranial pressure during the infusion period (10). Kroll et al. confirmed findings that showed a linear increase of VD proportional to increase in time, volume, and dose with a constant infusion rate and concentration (13). They studied the pharmacokinetics of iron oxide nanocompound (MION) and CED characteristics in a rat...

General Principles of Gene Regulation

Consider E. coli, a bacterium that resides in your large intestine. Your eating habits completely determine the nutrients available to this bacteria it can't seek out nourishment when nutrients are scarce nor can it move away when confronted with unpleasant changes. E. coli makes up for its inability to alter the external environment by being internally flexible. For example, if glucose is present, E. coli uses it to generate ATP if there's no glucose, it utilizes lactose, arabinonse, maltose, xylose, or any of a number of other sugars. When amino acids are available, E. coli uses them to synthesize proteins if a particular amino acid is absent, E. coli produces the enzymes needed to synthesize that amino acid. Thus, E. coli responds to environmental changes by rapidly altering its biochemistry. This biochemical flexibility, however, has a high price. Producing all the enzymes necessary for every environmental condition would be energetically expensive. So how does E. coli maintain...

Purification of Enzyme Product

Using a flash evaporator and a vacuum pump, enzyme product is exchanged three times with 99.99 deuterium oxide and analyzed by 1H NMR spectroscopy. This will indicate the linkage of the newly-added sugar. 3. For further linkage analysis, radioactive enzyme product is analyzed for its susceptibility to cleavage by galactosidases. The specificity of galactosidases helps to identify the linkage point and anomeric configuration of the newly-added sugar in the enzyme product. Incubate an aliquot (2000 cpm) of enzyme product at 37 C for 60 min with 0.01-1 U ( imol min) of a- or P-galactosidase in 100 L containing citric acid sodium phosphate buffer and 0.01 BSA at pH 4.3. After the incubation, add 800 L of water and pass the mixture through a C18 Sep-Pak column. Undigested product should elute with methanol as described in Subheading 3.3. Digested product elutes as free 3H galactose when eluted with water, and as non-radioactive substrate when eluted with methanol. A comparison of controls...

Carbon Sources in Environment

As the temperature begins to rise in a compost heap, the mesophilic microflora is succeeded by thermophilic microflora (Hedger and Hudson, 1974). Therefore, the availability of soluble carbon sources (sugars, amino acids and organic acids) will decrease and the carbon source available to thermophilic fungi will mainly be the polysaccharide constituents of the biomass, chiefly cellulose and hemicelluloses. Not surprisingly, thermophilic fungi are therefore especially well adapted for polysaccharide utilization. The growth rate of the thermophilic fungus Sporotrichum thermophile on cellulose (paper) is similar to that on glucose (Bhat and Maheshwari, 1987). Chaetomium thermophile and Humicola insolens grew better on xylan than on simple sugars (Chang, 1967).

Mixed Substrate Utilization

In composting plant material, the hydrolysis of polysaccharide constituents by the secreted enzymes is expected to produce a mixture of sugars in the growth environment of thermo-philic fungi. One of the adaptive strategies for their growth could be the simultaneous utilization of a mixture of sugars. To test this, the thermophilic fungi Thermomyces lanug-inosus and Penicillium duponti were grown in a mixture of glucose and sucrose in liquid media (Maheshwari and Balasubramanyam, 1988). Both fungi concurrently utilized glucose and sucrose at 50 C, with sucrose utilized faster than glucose (Figure 10.6). This is quite the opposite of the phenomenon of diauxy observed in bacteria that utilize one carbon source at a time, e.g., glucose is utilized before lactose in a mixture of the two sugars. The simultaneous utilization of sucrose in the presence of glucose occurred because invertase is insensitive to catabolite repression by glucose and because the activity of the glucose uptake...

Positive Control and Catabolite Repression

E. coli and many other bacteria will metabolize glucose preferentially in the presence of lactose and other sugars. They do so because glucose enters glycolysis without further modification and therefore requires less energy to metabolize than do other sugars. When glucose is available, genes that participate in the metabolism of other sugars are repressed, in a phenomenon known as catabolite repression. For example, the efficient transcription of the lac operon takes place only if lactose is present and glucose is absent. But how is the expression of the lac operon influenced by glucose What brings about catabolite repression

Products Available

Cranberry is available in a variety of forms such as fresh or frozen cranberries, cranberry juice cocktail, other cranberry drinks, cranberry sauce, and powder in hard or soft gelatin capsules (2,10). Cranberries are approx 88 water and contain flavonoids, anthrocyanins (odain), cetechin, triterpinoids, y-hydroxybutyric acid, citric acid, malic acid, glucuronic acid, quinic acid, benzoic acid, ellagic acid, and vitamin C (2). Fresh or frozen cranberries are a good source of cranberry because they contain pure fruit however, because of their high acidity and extremely sour taste, they are less readily used in clinical practice (1). Pure cranberry juice is tart like lemon juice because of the high citric and quinic acid content (2). Cranberry juice cocktail is more palatable, but is only 25-33 juice and contains corn syrup as a sweetener (2,10), whereas other cranberry juice drinks contain as little as 10 juice (2). These sweetened beverages are relatively high in calories (approx 140...

Isabell A Schmitt Milana Dolezal Michael F Press

The initial in vivo human studies demonstrating a correlation between estrogen uptake by the tumor tissue and responsiveness to hormonal ablation therapy were followed by in vitro observations in primary breast cancer slices or ho-mogenates. Procedures for cell fractionation and sucrose density gradient analysis were developed, which permitted the identification and quantification of estrogen binding in breast cancer samples. Using these techniques, the characteristics of an estrogen receptor (ER) were defined as a protein which bound its estrogen ligand in a highly specific fashion, with high affinity (Kd of 10 10 M) and with saturation by excess ligand. This interaction with the receptor protein was considered to mediate a variety of biological responses. Use of in vitro assays of tumor tissue samples improved the predictions of patient response to hormonal therapy. Approximately 50 -60 of women with ER-rich breast cancers responded to such therapy, while fewer than 10 with ER-poor...

Production and purification of exosomes

Since exosomes mediate MHC class I- and class II-restricted T-cell stimulatory capacity (29-31) and efficiently substitute for whole DC cultures (5), good manufacturing process (GMP) laboratory procedures for exosome harvesting and purification have been set up for clinical implementations (32). Exosomes derived from DC and tumor cell culture supernatants can be readily purified within 4-5 h starting from 2-3 liters of culture supernatant based on their physical properties. Exosomes float on sucrose gradients at a density ranging from 1.13 (for B-cell-derived exosomes) to 1.210 g cm3 (for dexosomes texosomes texas) (4,6,11,25,33,34). Ultrafiltration of the clarified culture supernatant through a 500-kDa hollow fiber membrane followed by ultracentrifugation onto a 30 sucrose deuterium oxide cushion (density 1.13-1.210 g cm3) reduced the volume and protein concentration approx 200-fold and 1000-fold, respectively. The percentage recovery of exosomes is about 40-50 , based on the...

What Are Carrot Shaped Brain Tumors

High Grade Large Cell Malignancy

Atypical teratoid rhabdoid tumors (WHO grade IV) are rare neoplasms that usually develop in the posterior fossa and affect children less than 3 years of age 24,25,30 . Histological examination reveals a cellular tumor with large and polygonal rhabdoid cells with conspicuous eosinophilic or pink cytoplasm, that contain spherical fibrillary intracytoplas-mic inclusions composed of bundles of intermediate filaments 51 . Other tissue elements may be present, such as primitive neuroepithelium, mesenchyme, and mature epithelium. Mitoses and regions of necrosis are common. The rhabdoid cells lack true muscle differentiation and do not react positively with desmin. They may show variable reactivity with epithelial-membrane antigen, vimentin, smooth muscle actin, and GFAP. These tumors have marked proliferative capacity as shown by Ki-67 studies, with labeling indices ranging from 50 to 80 per cent. Molecular studies consistently reveal a loss of chromosome 22 or specific loss of 22q11, with...

The Behavioral Environment

The family environment has the potential to promote healthy eating. For example, children who eat meals with their family consume more fruits and vegetables, have a more nutritionally dense diet with less of their energy intake derived from fat, and drink fewer carbonated and sugared beverages than do children who do not eat with their families (101). A recent summary of the intakes of youth ages 2-19 years, based on the U.S. Department of Agriculture's 1989-1991 Continuing Surveys of Food Intakes by Individuals, documented that American children have a widespread need for improvement in their diets (102). Indeed, the authors found that only 1 of youth met their recommended intake of nutrients, and membership in this small group was associated with excess intake, especially of fat (102). Daily consumption of three meals of approximately equal energy content may be another potential strategy to reduce adiposity in children (103). Further evidence that food patterns may contribute to...

Absorption in the Small Intestine

Carbohydrate digestion begins in the mouth with the activity of salivary amylase and is completed in the small intestine by enzymes from the intestinal mucosa and pancreas. The resulting monosaccharides are absorbed by the villi and enter blood capillaries (fig. 17.41). Simple sugars are absorbed by facilitated diffusion or active transport (see chapter 3, pp. 84 and 88).

Mechanisms Of The Bifidogenic Effect

Interestingly, while many bifidobacteria grow well when cultured with prebiotic oligosaccharides as their sole carbon and energy source, they often do not grow when supplied only with the monosaccharides from which these oligosaccharides are composed (74,171,172). This physiology may be another consequence of their evolution in an environment with a limited availability of simple sugars. It suggests that bifidobacteria lack transport mechanisms for many monosaccharides and import prebiotic oligosaccharides before hydrolyzing and metabolizing them. This presumably minimizes the availability of released simple sugars for cross-feeding by other intestinal bacteria and may be another factor contributing to the specific bifidogenic effect of NDOs.

Tcc Receptors And Signal Transduction

Lipid raft isolation protocol Lipid rafts are insoluble in nonionic detergents such as Triton X-100, and application of a sucrose gradient (45 to 5 sucrose) allows them to be separated from the other membrane domains. For this purpose, cell lysates in Triton X-100 1 are overlaid with sucrose and ultra-centrifuged for 16 h at 100,000 rpm at 4 C. Fractions from the top are collected and analyzed for lipid raft markers, among the best of which are gangliosides (GM1 and GM3). However, recent data indicate heterogeneity in lipid raft composition in each cell type. In this connection, flotillin can be used, even for functionally different rafts. Once the lipid raft fractions are identified, several experiments including protein localization phosphorylation (Western-blotting), kinase activity, and immunoprecipitation can be performed to assess composition. Lipid raft fluidity can be assessed using diphenylhexatriene as a probe for fluorescence anisotropy.

Iipolysaccharides And Product Development

Figure 1 gives a simplified product development scheme. The raw material may come from seeds (e.g., guar, locust bean gum, corn, wheat starch), tubers and roots (e.g., potato and tapioca starch, konjac), fruits (pectin), seaweeds (e.g., agar, alginate, car-rageenan, furcellaran), plant exudates (e.g., ghatti, gum arabic, karaya, tragacanth), and various plant sources (cellulose), or microorganisms (dextran, gellan, pullulan, xanthan). These substances are collected through appropriate means, cleaned, purified, and often modified. The modification may be chemical (e.g., to form substituted celluloses or modified starches), or microbial (e.g., to convert sucrose to dextran).

Plasma Membranes Are Involved In Facilitated Diffusion Active Transport Other Processes

Transport systems can be described in a functional sense according to the number of molecules moved and the direction of movement (Figure 41-10) or according to whether movement is toward or away from equilibrium. A uniport system moves one type of molecule bidirectionally. In cotransport systems, the transfer of one solute depends upon the stoichiometric simultaneous or sequential transfer of another solute. A symport moves these solutes in the same direction. Examples are the proton-sugar transporter in bacteria and the Na+ -sugar transporters (for glucose and certain other sugars) and Na+-amino acid transporters in mammalian cells. Antiport systems move two molecules in opposite directions (eg, Na+ in and Ca2+ out). A discussion of the transport of glucose summarizes many of the points made in this chapter. Glucose must enter cells as the first step in energy utilization. In adipocytes and muscle, glucose enters by a specific transport system that is enhanced by insulin. Changes in...

Isolation of Fruit Quality QTL

Correspondence of map location of QTLs and genes related to carbon metabolism allowed the identification of several putative candidate genes (Causse et al. 2004), but the validation of their role in trait variation required fine-mapping and the identification of a causal polymorphism that was not obvious. Mutations of enzymes involved in carbon metabolism that alter sugar composition in fruit have been found in S. chmielewskii and in S. habrochaites. The sucr mutation in an invertase gene in S. chmielewskii provides fruit with sucrose instead of glucose and fructose (Chetelat et al. 1995). In S. habrochaites, an allele of the ADP glucose pyrophosphorylase enzyme was identified as being much more efficient than the allele of cultivated tomato, leading to an increase in the final sugar content of fruit (Schaffer et al. 2000).

Soil Testing For Detecting Plant Nutritional Needs

Critical nitrate levels of sugar beets estimated from analysis of petiole and blades, with special reference to yields and sucrose concentrations. Soil Sci. 69 291-309, 1949. A Ulrich. Plant tissue analysis as a guide in fertilizing crops. In H.M. Reisenhauer, ed. Soil and Plant Tissue Testing in California. Riverside University of California Bulletin 1976, 1879, pp. 1-4. H. Mills, J.B. Jones, Jr. Plant Analysis Handbook II. Athens, Ga. Micro Macro Publishing, Inc., 1996, p. 422.

Digestion Absorption Of Carbohydrates

The digestion of complex carbohydrates is by hydrolysis to liberate oligosaccharides, then free mono- and di-saccharides. The increase in blood glucose after a test dose of a carbohydrate compared with that after an equivalent amount of glucose is known as the glycemic index. Glucose and galactose have an index of 1, as do lactose, maltose, isomaltose, and trehalose, which give rise to these monosaccharides on hydrolysis. Fructose and the sugar alcohols are absorbed less rapidly and have a lower glycemic index, as does sucrose. The glycemic index of starch varies between near 1 to near zero due to variable rates of hydrolysis, and that of non-starch polysaccharides is zero. Foods that have a low glycemic index are considered to be more beneficial since they cause less fluctuation in insulin secretion. bifunctional enzyme catalyzing hydrolysis of sucrose and isomaltose), lactase, and trehalase are located on the brush border of the intestinal mucosal cells where the resultant...

Plate 87 Seminal Vesicle

The secretion of the seminal vesicles is a whitish yellow viscous material that contains fructose, other simple sugars, amino acids, ascorbic acid, and prostaglandins. Although prostaglandins were first isolated from the prostate gland (hence the name), they are actually synthesized in large amounts in the seminal vesicles. Fructose is the primary nutrient source for the sperm in the semen.

Transcription Sites and Nuclear Territories Functional Organization ofInterphase Nuclei

Using chromatin precipitation, sucrose gradient sedimentation, and array comparative genomic hybridization (aCGH) techniques, Gilbert et al. (86) showed that gene-rich domains are enriched in open chromatin fibers and suggested that domains of open chromatin may create an environment that facilitates transcriptional activation and could provide an evolutionary constraint to maintain clusters of genes together along chromosomes.

Boon Huan Tan Jian Lin Fu and Richard J Sugrue

The full-length and truncated forms of recombinant envelope (E) glycoprotein from Dengue virus type 1, Singapore strain S275 90 were expressed in the yeast, Pichia pastoris, using a secretory vector. A truncated form of the E protein in which the transmembrane domain was deleted was secreted successfully into the culture medium. The E protein was also co-expressed with C and prM proteins using a non-secretory yeast vector. The co-expression of C, prM and E proteins resulted in the spontaneous formation of virus-like particles (VLPs), which were confirmed by sucrose gradient analysis and transmission electron microscopy. Furthermore, the VLPs were used to immunise rabbits, and shown to be immunogenic by immunofluorescence staining of dengue virus-infected Vero cells. The yeast-expressed E protein was treated with PNGase F, which showed that although the protein was modified by the addition of W-linked glycans, the recombinant expressed E protein was not hyperglycosylated. Key Words...

Former Reader in Pharmacognosy University of Bradford UK

A decade later Professor Kondo's research team studied aqueous, methanolic and ethereal extracts of ginseng roots. The aqueous fraction yielded mucilage and inorganic compounds, the methanolic extract sucrose, some nitrogenous substances and a saponin glycoside, and the ether extract an oily material. Steam distillation of this oily fraction yielded two products, a light, yellowish volatile oil and a brown non-volatile residue. From the volatile oil the camphor-like panacene was recovered, and the non-volatile fraction contained phytosterol and fatty acids. Study of the saponin fraction, using 7 per cent alcoholic hydrochloric acid as the hydrolysing agent, produced two compounds, a crystalline panax-sapogenol and an amorphous panaxsapogenol. Closer investigation confirmed that the saponin glycoside had a molecular weight of 876 and comprised a molecule of panax-sapogenol linked to two molecules of glucose and one molecule of pentose. Kondo's team also observed that cultivated...

Eight Sugars Predominate In Human Glycoproteins

About 200 monosaccharides are found in nature however, only eight are commonly found in the oligosaccharide chains of glycoproteins (Table 47-4). Most of these sugars were described in Chapter 13. -Acetyl-neuraminic acid (NeuAc) is usually found at the termini of oligosaccharide chains, attached to subterminal galactose (Gal) or -acetylgalactosamine (GalNAc) residues. The other sugars listed are generally found in more internal positions. Sulfate is often found in glyco-proteins, usually attached to Gal, GalNAc, or GlcNAc.

Formation of Virus Like Particles

Sucrose Gradient Centrifugation 3. Prepare a gradient in the centrifuge tube from 5 to 50 of sucrose solutions made up in PBS (see Note 4). 1. Place 10 L of each sucrose fraction onto the EM grid. Drain dry. To determine whether the co-expression of C, prM, and E could form VLPs, the yeast transformmant containing the CprME construct was induced, the cells lysed, and the clarified lysate applied to a 5 to 50 sucrose gradient, which was centrifuged for 16 h at 100,000g (Fig. 5A). Our results indicate that fraction 3, collected from the top of the gradient, indicates the presence of E protein when analysed by Western blotting with the antiserum raised to bacterial-expressed E protein (lane 3). When the peak fraction 3, was further analyzed by TEM, VLPs were observed at a high magnification of X60,000 (Fig. 5B). Fig. 5. Biophysical analysis of recombinant E proteins. Protein expression in the yeast transformant containing the CprME construct was induced, the cells lysed and the...

Major and satellite signals

We should perhaps add that all the signalling molecules we have considered are essentially plant produced and plant orientated, but our consideration is not exhaustive. For example, polyamines are common to both eukaryotes and prokaryotes and as low molecular weight compounds they can regulate and influence metabolic events (Kumar et al., 1997). Sugars (sucrose or glucose) levels in a cell operate a complex signalling network with ethylene and abscisic acid to inhibit or promote specific growth responses (Leon and Sheen, 2003). Such substances are universal signal molecules. Perhaps as every cell is a target cell, so, given the appropriate circumstances, every molecule can become, at least temporarily, a signal molecule.

Overview Of Metabolic Substrate Engineering

The complex glycoconjugates expressed on eukaryotic cell surfaces (Figs. 3-8) comprise primarily 10 monosaccharides Glc, Gal, Man, Fuc, GlcN, GlcNAc, GalNAc, Xyl, GlcUA, and sialic acid (Neu5Ac). These sugars can be biosynthesized de novo within a cell, often from Glc or, in many cases, key metabolic intermediates can be supplied exogenously to a cell. The pathways for these interconversions are summarized in Figure 9 33-36 . If the enzymes in these biosynthetic pathways could tolerate unnatural substrates, subtle modifications, such as replacement of a hydroxyl group with a hydrogen atom or halogen, could be introduced into cell surface gly-cans. This possibility attracted the attention of many groups interested in understanding how specific carbohydrate structures dictate molecular and cellular interactions.

Nonwestern Medical Systems Traditional Chinese Medicine

Traditional Chinese Medicine (TCM) has existed for thousands of years, long before Western medicine. Rather than following the disease model of Western medicine, TCM focuses on a symptom approach such that a person with PD who has mostly tremor would be evaluated and treated differently than another person whose symptoms were mostly gait and balance difficulty with no tremor. The specific symptoms of the individual signal a deficiency in the body fluids blood that is unable to properly nourish the energy flow or chi or Qi of the entire organism. There are three main symptom approaches under TCM (5). The first is Qi and blood deficiency, which is believed to arise from anger, emotional stress, frustration, and resentment. The second is phlegm-fire-agitating wind (yang), which is the result of poor diet, in particular eating greasy, fried, sweet, sugary foods and alcohol. The third is kidney and liver (yin) deficiency, which results from a lack of rest and overwork as well as part of...

Origins of celltocell signalling

Traces of sucrose have been shown to increase the production of ethylene and stimulate xylogenesis in lettuce pith explants (Warren-Wilson et al., 1994), and so it is interesting to speculate that the transportation of inductive concentrations of sucrose along the conductive tissues of the phloem in higher plants and along the elongated cell pathways carrying metabolites in primitive plants could have been one of the causal signals to the evolutionary development of lignin-like elements in the plant body.

Secretions of the Small Intestine

Epithelial Cell Small Intestine

The fluid the intestinal glands secrete has a pH that is nearly neutral (6.5-7.5), and it lacks digestive enzymes. However, the epithelial cells of the intestinal mucosa have digestive enzymes embedded in the membranes of the microvilli on their luminal surfaces. These enzymes break down food molecules just before absorption takes place. They include peptidases, which split peptides into their constituent amino acids sucrase, maltase, and lactase, which split the double sugars (disaccharides) sucrose, maltose, and lactose into the simple sugars (monosaccharides) glucose, fructose, and galactose and intestinal lipase, which splits fats into fatty acids and glycerol. Table 17.9 summarizes the sources and actions of the major digestive enzymes.

Mechanisms of Interferon Action

Interferon is known to activate the proteasome, which is a ubiquitously expressed multi-subunit complex that degrades proteins. To examine the possibility that IFN treatment induced the proteasome that specifically degrades GFP translated from the IRES construct, inhibition of GFP expression from IRES clones by interferon was assessed in the presence of two proteasome inhibitors lactacystin and epoxomicin (Fenteany & Schreiber, 1998 Meng et al., 1999). Inhibition of GFP expression from the IRES clones by IFN treatment was not affected when Huh-7 cells were pretreated with the proteasome inhibitors, indicating that interferon action on the IRES inhibition is not due to activation of proteasome pathways. We also examined the possibility that interferon treatment impaired the loading of polyribosome with IRES containing mRNA in the transfected Huh-7 cells. This was accomplished by examining the distribution of IRES containing GFP mRNA in polysome fractions by northern blot analysis....

Human Flora Associated Animals

HFA animals are created by inoculating germ-free animals with a human fecal homogenate (94). The resulting microbial profile of HFA animals is partly dependent on the differing ability of the various microorganisms in the human fecal sample to colonize the animal GIT. Previous studies have shown that certain microorganisms of human fecal origin were unable to colonize the rodent GIT (95). There may be several reasons for this, such as diets or host factors like transit times and physiological conditions. It has been demonstrated that mice, fed with a commercially available animal feed, may have a reduced, or even undetectable level of bifidobacteria in feces. However, after feeding these mice an alternative diet for several weeks, bifidobacteria could be detected in the mice that were fed sucrose or amylose, with particularly dense populations of bififodobacteria

Natural And Organic Spice Trends

Thus chemicals, pesticides, or fumigants are not used during spices' growth, during their storage, or to eliminate microbial growth. Organic spices contain none of the fillers (sucrose, starch, or dextrose), synthetic anti-caking agents, artificial colors and flavors, or preservatives that may be found in conventional spices. Many spice-producing countries are already in the organic spice-production trade, including the United States, India, Guatemala, Turkey, Indonesia, and Egypt.

Ingestion Of Large Quantities Of Fructose Has Profound Metabolic Consequences

Diets high in sucrose or in high-fructose syrups used in manufactured foods and beverages lead to large amounts of fructose (and glucose) entering the hepatic portal vein. Fructose undergoes more rapid glycolysis in the liver than does glucose because it bypasses the regulatory step catalyzed by phosphofructokinase (Figure 20-5). This allows fructose to flood the pathways in the liver, leading to enhanced fatty acid synthesis, increased esterification of fatty acids, and increased VLDL secretion, which may raise serum triacylglycerols and ultimately raise LDL cholesterol concentrations (Figure 25-6). A specific kinase, fructokinase, in liver (and kidney and intestine) catalyzes the phosphorylation of fructose to fructose 1-phosphate. This enzyme does not act on glucose, and, unlike glucoki-nase, its activity is not affected by fasting or by insulin, which may explain why fructose is cleared from the blood of diabetic patients at a normal rate. Fructose 1-phos-phate is cleaved to...

Pharmacological Toxicological Effects

There are numerous accounts of anecdotal evidence supporting the use of St. John's wort for premenstrual syndrome (PMS) (36). One open, uncontrolled study was conducted to determine the efficacy of St. John's wort in treating PMS. The primary outcome was measured by a daily symptom checklist of 17 symptoms rated on a scale of 0 to 4 based on the Hospital Anxiety and Depression (HAD) scale and modified Social Adjustment Scale (SAS-M) broken down into four subscales mood, behavior, pain, and physical. A total of 25 women were selected to participate in the study in which they received 300 mg hypericum standardized to 900 g hypericin daily. The results from the daily symptoms survey after the first cycle show a statistically significant reduction from the baseline value of 128.42 to 70.11. After the second cycle, there was a further reduction to 42.74. Of the four subscales, St. John's wort had the greatest improvement on the mood subscale (57 ) and the least improvement on the physical...

In Vivo Expression Technology Approaches

Nine of the 72 ivi genes appeared to encode sugar-related functions, including genes involved in ribose, cellobiose, sucrose, and sorbitol transport. Another nine genes encode functions involved in acquisition and synthesis of amino acids, nucleotides, cofactors, and vitamins, indicating their limited availability in the GI tract. Four genes involved in stress-related functions were identified, reflecting the harsh conditions that L. plantarum encounters in the GI tract. Another four genes encoding extracellular proteins were identified that could mediate interactions with host GI-tract epithelial cells. Remarkably, the protein encoded by one of the hypothetical proteins identified in this study in L. plantarum is a homologue (32 identity) of the only conserved hypothetical protein that was identified with IVET in L. reuteri (43). Moreover, a large number of the functions and pathways identified in L. plantarum have previously been identified in pathogens as...

Phase I Trial Using Human DCDerived Exosomes

These data were the rationale for launching vaccination with DC-derived exosomes in metastatic tumor-bearing patients. A GMP has been set up to harvest large amounts of MD-DC-derived exosomes (ultrafiltration followed by ultracentrifugation on a high-density sucrose D2O cushion) and load exosomal MHC class I molecules with synthetic tumor peptides. Phenotypical analysis of clinical-grade exosomes is allowed by FACS-beads-assay, i.e., DC-derived exosomes, are fixed onto 4.5-micron beads stained using surface antigen-recognizing fluorescent antibodies. An immunocapture assay determines the quantity of MHC class I and or II molecules in each individual exosome batch. A feasibility and safety phase I study was undertaken in France (Institut Gustave Roussy and Institut Curie with Anosys Inc. biotechnological partnership) in HLA-A1 B35 and -DP04 patients bearing stage III IV melanoma expressing MAGE-3. Dexosomes were purified from the culture supernatant of day 7 autologous MD-DCs. MAGE-3...

Orphan Gpcrs In The Central Nervous System

These peptides were initially shown to exert activity in appetite control, but orexin-knockout mice soon displayed disrupted sleep patterns similar to narcolepsy.198 At the same time, positional cloning in a narcoleptic dog colony revealed that the canarc-1 gene corresponds to the orexin B receptor.199 The autosomal recessive canarc-1 mutation in Doberman pinschers and Labrador retrievers produces narcolepsy that is pharmacologically and physiologically validated with the human condition. The easily detectable activity of orexin peptides has substantiated interest in their receptors as potential CNS drug targets. However, other GPCRs with unknown or less well-characterized ligands represent major challenges for drug discovery.200

Immunogenicity of Peritoneal Ascitis Derived Exosomes

Tumor-derived exosomes are not simply released in vitro by tumor cell lines in culture supernatants. We examined malignant effusions for the presence of tumor-derived exosomes and analyzed exosome immunogenicity on autologous peripheral T lymphocytes. Ultracentrifugation on sucrose and D2O gradients of11 malignant effusions allowed isolation of abundant amounts of exosomes. Malignant effusions accumulate high amounts of membrane vesicles with a mean diameter of 60-90 nm. These vesicles bear antigen-presenting molecules (MHC class I MHC class II, heat shock proteins), tetraspanins (CD81), and contain tumor antigens (Her2 Neu, MART-1, TRP, gp100). Up to 2 x 1014 exosome-associated MHC class I molecules are recovered from malignant ascitis of 2-3 liters. Exosomes from melanoma patients shuttle MART-1 tumor antigen to MD-DCs for cross-presentation to MART-1-specific CTL clones. In seven of nine cancer patients, tumor-specific lymphocytes could be efficiently expanded from peripheral blood...

Chromogenic Reactions

Many different enzymes produce the same molecules (e.g., NADH, NADPH, orthophosphate, ammonia, hydrogen peroxide, etc.) or different molecules with essentially the same chemical properties (e.g., aldehydes, ketones, reducing sugars, thiols, etc.). This means that very similar or identical chromogenic reactions can be used to detect different enzymes. Thus, classification of chromogenic reactions based on properties of products that are detected seems to be the more useful and practical approach. Such classification is advantageous because it allows one to choose an adequate chromogenic reaction to visualize activity bands even of those enzymes that have not yet been detected on electrophoretic gels.

Treatments

The National Center for Complementary and Alternative Medicine (NCCAM) is conducting an ongoing series of clinical tests of alternative and complementary treatments for depression. Those that have been shown to reduce symptoms of depression and compare favorably with conventional treatments include acupuncture Ayurvedic medicine meditation and a therapeutic diet designed to be free of caffeine and refined sugar.

Carbohydrates

Carbohydrates are water-soluble molecules that contain atoms of carbon, hydrogen, and oxygen. These molecules usually have twice as many hydrogen as oxygen atoms, the same ratio of hydrogen to oxygen as in water molecules (H2O). This ratio is easy to see in the molecular formulas of the carbohydrates glucose (CbH Ob) and sucrose (C12H22O11). Carbohydrates are classified by size. Simple carbohydrates, or sugars, include the monosaccharides (single sugars) and disaccharides (double sugars). A monosac-charide may include from three to seven carbon atoms, occurring in a straight chain or a ring (fig. 2.10). Mono-saccharides include glucose (dextrose), fructose, and galactose. Disaccharides consist of two 6-carbon units. Sucrose (table sugar) and lactose (milk sugar) are disac-charides (fig. 2.11a and b).

Nonparental ditype

It is important to note that the phenotype of the double mutant may be unique. This occurs most often when the two genes encode functions involved in the same process. If mutant strains containing alterations in unrelated gene functions, such as ade2 and suc2, are crossed, then the double mutant is expected to exhibit both phenotypes, adenine requiring and unable to utilize sucrose. If mutant strains containing alterations in two related gene functions are constructed, such as MCM2 and MCM7 encoding different components of the origin recognition complex (ORC), then the double mutant could exhibit an unexpected phenotype. For example, the double mutant combination could be lethal even though each single mutant strain is viable. Often mutant genes are crossed for the purpose of determining the double mutant phenotype. As will be discussed in detail in the chapters on epistasis, suppression, and enhancement, a great deal of insight into the function and relationship between gene products...

Catabolism

The hydrolysis of a disaccharide, for instance, results in two monosaccharide molecules (see fig. 4.1 read from right to left). In this case, the bond between the simple sugars breaks, and the water molecule supplies a hydrogen atom to one sugar molecule and a hydroxyl group to the other. Thus, hydrolysis is the reverse of dehydration synthesis.

Cell Fractionation

Equilibrium density gradient centrifugation separates subcellular components based only on their density. For this method, one must first prepare a density gradient in a centrifuge tube. A nonionic molecule like sucrose, glycerol, or Ludox is used to vary the density of the buffer solution. The concentration of the molecule is varied, and therefore the density of the solution, and the concentration, is greatest at the bottom of the centrifuge tube and decreases slowly towards the top of the tube. Special devices are available for making these gradients. A step gradient can also be prepared. Here a series of solutions of different concentration (30 , 25 , 20 , etc.) are layered on top of one another with the step with the highest concentration at the bottom.

Scheme

Toshima et al. found that unprotected 2-deoxy sugars, as either reducing sugars or methyl glycosides, will undergo C-glycosidation to give aryl C-glycosides 158 . The reaction has been catalyzed by TMSOTf, and it has also been found that montmorillonite K-10, used in excess, also works with unprotected sugars (Scheme 87) 159 . Work by Park et al. deals with regioselective acylation of the 2'-hydroxyl group on a free aloesin glucopyranoside (not shown) 160 .

Ghsr Expression

The expression patterns of the type 1a and type 1b GHS-Rs were studied by ribonu-clease protection analysis in human and rat tissues and by in situ hybridization histochemistry in rhesus hypothalamus and rat brain and pituitary. Functional assessment of sucrose gradient-fractionated poly (A)+ mRNA from swine pituitary gave a single peak of GHS-R activity in the size range 1.6-2.3 kb (Fig. 7). However, attempts at detecting GHS-R mRNA by Northern blotting analysis have been unsuccessful, even though control mRNAs for other GPC-Rs could easily be detected. The authors attribute the difficulty in detecting GHS-R mRNA by Northern blotting analysis to its low abundance and potential size heterogeneity. TRH and GnRH receptors were also readily detected functionally. PCR amplification of the swine pituitary GHS-R cDNA sequences from among 11 pools of an unamplified pituitary cDNA library (110,000 individual cDNAs pool) resulted in GHS-R cDNA identification in only 4 of 11 pools. Therefore,...

Food Source

Cocoa and chocolate are nutritious foods that contribute to caloric as well as trace mineral intake (Steinberg et al 2003). Milk chocolate has a relatively low glycaemic index of approximately 40 (Foster-Powell et al 2002) and this is attributed to the fat in chocolate slowing gastric emptying and thus the rate of subsequent digestion and absorption. The glycaemic effect of milk chocolate can be further reduced by replacing the sucrose with fructose or isomalt (Gee et al 1991). Foods containing cocoa have been shown to lead to a greater postprandial insulin secretion in healthy young adults than foods with alternative flavourings, despite having a similar glycaemic index. It is suggested that specific insulinogenic amino acids or greater cephalic phase insulin release may explain this finding, although the clinical implications are uncertain (Miller et al 1995).

Reaction

0.1 M Tris-HCl buffer, pH 8.8 60 mM Indole-3-acetaldehyde 5 mM NAD 20 Sucrose The addition of PMS and MTT (or NBT) to the staining solution will allow development of IADH activity bands visible in daylight. In some cases the addition of PMS and a tetrazolium salt should be made only when fluorescent IADH bands visible under UV light are well developed. This postcoupling technique allows avoidance of the possible inhibitory effect of PMS or a tetrazolium salt on IADH activity. When a tetrazolium method is used, sucrose may be omitted from the staining solution.

Let there be light

In nature, plants depend upon the energy of the sun. Through a process called photosynthesis, sunlight is converted to sugars to provide fuel for growth. These sugars are utilized as necessary in a process called respiration, excess sugar can then be stored for later use. Photosynthesis is made possible by chlorophyll which is contained within the leaf cells. It is this chlorophyll which gives vegetation its characteristic green color. Light is trapped by the chlorophyll, activating the process of photosynthesis. Inside the chlorophyll, light energy is combined with carbon dioxide and water to produce oxygen and sugar. The sugar is then oxidized (or metabolized) through the process of respiration, producing carbon dioxide, water, and energy for growth. Excess oxygen and water are transpired by the leaf into the air. Plant growth, therefore, is directly affected by the amount and quality of light it receives.

Fisher Glycosylation

Furanose glycosides of simple alcohols are conveniently obtained by subjecting a reducing sugar to a controlled Fisher glycosylation reaction. The furanosides are the kinetic products of this reaction, and excellent yields of the product (as a mixture of anomers) are usually obtained if the proper conditions are chosen 67 . The alcohol generally used in this reaction is methanol, which serves as both a reactant and the solvent. The methyl glycosides thus obtained must then be protected and converted to a suitable glycosylation agent. It is also possible to synthesize glycosides of other alcohols by this method however, since the alcohol is usually employed as the solvent, the choice is somewhat limited. Therefore, from a practical point of view, this method is most useful for the preparation of glycosides of relatively volatile alcohols (e.g., methanol, ethanol, and allyl alcohol).

Culture Conditions

Both rich and synthetic minimal media are used to culture Saccharomyces. Rich medium, called YEP or YP, is made from commercially available yeast extract and peptone (a complex protein digestion product). It contains all essential nutrients including ammonia (a rich nitrogen source), phosphate, sulfate, sodium, magnesium, calcium, copper, iron, etc. and certain other compounds that all Saccharomyces strains are unable to synthesize. In addition, rich medium provides many macromolecular precursors such as amino acids and nucleotides that wildtype Saccharomyces strains are able to synthesize if necessary. A sugar or other carbon energy source must be added, such as glucose (dextrose), sucrose, lactic acid, or others depending on the genotype of the strain and its ability to utilize various carbon sources. Glucose is the richest and most readily available carbon source and a rich medium containing glucose is referred to as YEPD or YPD. Because of the abundant nutrient supply, cells...

Nutrition

As the process of globalization occurs, so too does urbanization. Though the exact relationship may not be clear, it is obvious that these are related events. With globalization comes increasing 'modernization' of diets in large urban settings (Sobal, 1999). Modernization of global diets towards a Western model of eating is more common in cities around the world than it is in more rural, outlying areas. Therefore, although food may be highly available, it is also more processed, creating food stuffs that are likely to have higher ratios of simple sugars, 'bad' fats, and lower fiber content (Drewnowski, 1999).

Article

(a) Describe the method used to identify sucrose nonfermenting mutants. 4. Diagram the cross between two sucrose nonfermenting mutants isolated from strains DBY782 (MATa ade2 SUC2) and the otherwise isogenic strain DBY916 (MATa his4-86oc lys2-802oc) in which the mutations in the two strains are in the same complementation group (i.e. gene). Call the gene FER1 and show the genotype and phenotype of the mutant parents, the diploid, and the resulting tetrads.

Glycopolymers

Reducing sugars can be reductively aminated with ammonia or other amines. The resulting amine derivatives can then be transformed into -acrylamide monomers useful in copolymerization strategy. Unfortunately, acrylamide copolymeriza-tion affords polymers that may vary greatly in their batch-to-batch molecular weight distributions. An improved protocol consists of synthesizing polyacrylates (43) having active ester functionality NHS 83 or 4-NO2Ph 84 (Scheme 8). After aminolysis or hydrolysis, the molecular weight of the resulting polyacrylamides or polyacrylic acids can be compared against commercially available polymer standards used in HPLC. This allows the synthesis of reproducible lots of a given copolymer. Once established, the starting polyacrylates 43 can be treated with various amounts of any amine-containing sugars. Quenching the residual reactive esters with amine 1 (R1NH2) followed by amine 2 (R2NH2), or simply with ammonia or water, afforded copolymers having desired...

Conclusion

The purification process used for exosome isolation from DC cultures and ascites fluid allows harvesting of reproducible yields of 40- to 100-nm vesicles that are highly immunogenic in vitro. Exosomes are currently defined by their morphology (electronic microscopy), by their physical properties (stable at high temperature, floating at a density of 1.13-1.210 g mL in a sucrose D2O gradient), by their proteic patterns (endocytic markers, i.e., tetraspanins and hsp73), and by their enrichment in MHC class I, II, CD86 molecules when derived from DCs. Importantly, exosomes efficiently transfer antigens from APC to APC, allowing initiation and amplification of antigen-specific immune responses. Immunization of melanoma patients with DC-derived exosomes was associated with objective clinical responses and prompted the initiation of phase II trials. However, the physiological relevance of exosomes remains to be established and might ensue from the identification of the critical components of...

Animal Studies

The pressure gradient for the infusion is typically supplied by an external pump however, use of an internally implanted pump for chronic infusion has been reported 29 . The infusates used in the studies are often potentially therapeutic agents or tracer solutions, the latter of which are commonly labeled or radioactive, allowing researchers to study flow patterns and concentration gradients. The size of the solute can also vary by orders of magnitude, ranging from inert sucrose which is less than 1 nm in diameter to polystyrene or viral particles 200 nm in diameter, though the properties of convective delivery at this upper boundary remain ill-defined 19,26,33 . Nevertheless, it is clear from laboratory studies and human trials that protein and oligonucleotide solutes, with molecular diameters in the 5-20 nm range, are easily delivered and distributed in brain and tumor parenchyma using this approach.

Insect Resistance

S. pennellii accessions (particularly LA716) have shown a very high level of resistance to the whitefly Bemisia tabaci Bemisia argentifolii complex, as well as to aphids (Macrosiphum euphorbiae, Myzus persi-cae), mites and Lepidopteran pests (Gentile et al. 1968, 1969 Juvik et al. 1982 Goffreda et al. 1989), including the South American tomato pinworm Tuta ( Scrobi-palpuloides) absoluta (Franca et al. 1989). The multiple pest resistance of S. pennellii is due to the presence of type IV glandular trichomes and the glucose and sucrose esters of fatty acids (acylsugars) that they secrete (Gentile et al. 1968 Goffreda et al. 1989 Shapiro et al. 1994). A reduction of insects on plants has been shown using purified acylsugars, where feeding is reduced by Macrosiphum euphorbiae and Myzus persi-cae, a reduction in feeding, larval development and survival of Helicoverpa zea and Spodoptera exigua, and by reducing oviposition and feeding of the leaf miner Liriomyza trifolii and of the silver...

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Electrophoresis in free solution is not a common technique, for reasons that actually have little to do with the above considerations. Almost all electrophoresis is done with gels or other support matrices, even when the molecules involved do not behave as free draining. Why use a gel at all under these circumstances The reason is to prevent convective instabilities. Placement of an electrical field across a conductive solution leads to significant current flow and significant heating. This heating produces nonuniformities in temperature, and con-vective solvent motion will result from these, much as convection patterns are established if water is heated on the surface of a stove. The presence of bands of dissolved solute molecules leads to regions with local bulk density differences, as illustrated in Figure 5.3. These dense zones are gravitationally unstable. If the electrophoresis is carried out vertically, the dense zone can simply fall through the solution like a droplet of...

Figure 1630

Diagram showing the digestion and absorption of carbohydrates by an enterocyte. Carbohydrates are delivered to the alimentary canal as monosaccharides (e.g., glucose, fructose, and galactose), disaccharides (e.g., sucrose, lactose, and maltose), and as polysaccharides (e.g., glycogen and starch). Enzymes evolved in digestion of carbohydrates are classified as salivary and pancreatic amylases. Further digestion is performed at the striated border of the enterocytes by enzymes breaking down oligosaccharides and polysaccharides into three basic monosaccharides (glucose, galactose, and fructose). Glucose and galactose are absorbed by the enterocyte via an active transport utilizing Na+-dependent glucose transporters (SGLT1). These transporters are localized at the apical cell membrane (brown circles with G and Na' labels). Fructose enters the cell via facilitated NaHndependent transport utilizing GLUT5 (gray circle with F label) and GLUT2 glucose transporters (orange octagon with G2...

Addiction

Addiction has been postulated to play a role in disordered eating, with some individuals addicted to certain foods or combinations of foods. Although substance abuse and other impulse control disorders are associated with binge eating in some studies, there is no evidence that ''addiction'' to foods such as refined flour, simple sugars, or carbohydrates occurs or triggers binge episodes (71). An interesting finding, however, is that both lean and obese female binge eaters, women with binge eating disorder, who prefer to binge on foods that are both sweet and high in fat, may decrease their intake of these foods selectively when given naloxone (72). The role of h-endorphins and other endogenous opioids in the development or maintenance of binge eating, while unknown, is intriguing.

Invertase

Invertase, an enzyme from yeast studied extensively that played an important role in the development of biochemistry, is synthesized constitutively in the mesophilic fungi, wherein its activity increases with an increase in growth (biomass). By contrast, it is an inducible enzyme in the two thermophilic fungi studied, i.e., it is produced only in response to the availability of sucrose in the growth medium (Table 10.1). Strangely, the induced enzyme activity begins to diminish before any substantial quantity of sucrose is utilized or an appreciable amount of biomass is formed (Chaudhuri et al., 1999). Paradoxically, despite this pattern of development of invertase, the final mycelial yields are not affected. An investigation of this unusual pattern of development of invertase activity in a thermophilic fungus, T. lanuginosus, has given some insight into various strategies in the biochemical adaptation of thermophilic fungi.

Tissue Processing

Probably due to the localization of the satellite cells underneath the basement membrane of the muscle fiber, muscle specimens can be stored for a conveniently long period. If the tissue is kept on ice in an appropriate buffer, such as sterile Dulbecco's PBS (DPBS 145 mM NaCl, 5.4 mM KCl, 5 mM Na2HPO4, 25 mM glucose, 25 mM sucrose, 50 IU ml penicillin and 10 g ml streptomycin) 47 , or Ham's F12 medium supplemented with 10mM HEPES NaOH (pH 7.2) 44 , the tissue specimens can be stored before processing for at least three days. Slightly fewer viable cells will be isolated from the biopsies (Table 1), but storage has no effect on proliferation and differentiation rates. After three days, however, the cell yield seems to drop considerably 44 . This allows transportation over long distances, and considering the time-consuming procedure for the enzymatic isolation of satellite cells described below, better planning of the experiments.

Lectins

The agglutinating activity of lectins is inhibited by simple sugars, usually monosaccharides. It is assumed that these sugars represent the binding site for the lectin on the cell surface. Through the use of lectins, Morgan and Watkins 686 obtained some of the early information on the nature of the A, B, and H antigens. Some plant extracts contain more than one lectin, for example, seeds of Ulex europaeus and Bandeiraea simplicifolia (see below).

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Maffeis C, Provera S, Filippi L, et al. Distribution of food intake as a risk factor for childhood obesity. Int J Obes Relat Metab Disord 2000 24 75-80. Ludwig DS, Peterson KE, Gortmaker SL. Relation between consumption of sugar-sweetened drinks and childhood obesity a prospective, observational analysis. Lancet 2001 357 505-508. Nicklas TA, Elkasabany A, Srinivasan SR, Gerenson G. Trends in nutrient intake of 10-year-old children over two decades (1973-1994) the Bogulasa Heart Study. Am J Epidemiol 2001 153 969-977. Birch LL, Johnson SL, Andresen G, Peters JC, Schulte MC. The variability of young children's energy intake. N Engl J Med 1991 324 232-235. Fisher JO, Birch LL. Restricting access to palatable foods affects children's behavioral response, food selection, and intake. Am J Clin Nutr 1999 69 1264-1272. Johnson SL, Birch LL. Parents' and children's adiposity and eating style. Pediatrics 1994 94 653-661. Montgomery C, MacRitchie J, Jackson D, Reilly J. Relationship between...

Product Isolation

Ion exchange chromatography can be used for the isolation of sialylated products and oligosaccharides with amine groups. Size-exclusion chromatography serves well for the isolation of reducing sugars or glycoconjugates with high molecular weights. Adsorption on charcoal is another practical method for the purification of oligosaccharides. A recent publication focuses on the separation of oligosaccharides 132 .

The Sugar Solution

The Sugar Solution

Curb Sugar Cravings Once And For All With These Powerful Techniques. Sugar sensitive people might be low in specific neurochemicals that help us feel calm, centered, confident, and optimistic. Sugar is a drug that temporarily makes the sugar sensitive feel better, but with damaging consequences.

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