Natural Remedies for Food Cravings

Sugar Belly Secret

Joe Bovino is not only the creator of this book of strategies but also the author of other four amazon number one bestsellers. Having done extensive research and consulted professionals, he has formulated a strategy on how to get rid of that extra fat and lose weight. That is after almost a decade and a half year. He has also experienced other products prior to researching the natural ways of having a good strategy for weight loss. He can, therefore, be trusted. It entails a fun and simple strategy of having weight loss that melts away the extra pound without exercise or dieting. At times, it is quite hard to stay motivated to work out on a daily basis, especially when you are busy with work and getting older, it is hard to find the time and maintain your workouts! With this book of strategies, you learn how to continue with your usual work and enjoy life with your friends and family while at the same time lose that extra weight and belly, without any shed of sweat. It will help you; Rejuvenating and refreshing your skin, Supercharge your energy levels and become activated most of the time, You will still continue eating your preferred food and drinks without restrictions., Melt away extra pounds and keep them off for a long time. Read more here...

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Chromosome Specific DNA from Sucrose Gradients

Pure, minichromosome-sized DNA could also be obtained by sucrose-gradient fractionation (13)', a modification of the method by Sloof et al. (62 see also Note 9). 1. Make a 35 mL linear, 5-20 sucrose gradient in 100 mMEDTA, 25 mMTris-HCl, pH 7.5,1 SLS, and 50 Hg mL proteinase K in SW 28 ultracentrifuge tubes.

Aerobic Respiration Yields about 60 Molecules of ATP per Molecule of Sucrose

The complete oxidation of a sucrose molecule leads to the net formation of per sucrose by oxidative phosphorylation. The result is a total of about 60 ATPs synthesized per sucrose (Table 11.2). Using 50 kJ mol-1 (12 kcal mol-1) as the actual free energy of formation of ATP in vivo, we find that about 3010 kJ mol-1 (720 kcal mol-1) of free energy is conserved in the form of ATP per mole of sucrose oxidized during aerobic respiration. This amount represents about 52 of the standard free energy available from the complete oxidation of sucrose the rest is lost as heat. This is a vast improvement over the conversion of only 4 of the energy available in sucrose to ATP that is associated with fermentative metabolism.

Crop Plants And Renewable Resources

Potato lines with structurally modified starch (changes in amylose amylopectin ratios), rape transgenic genotypes accumulating seed oil with other than the normal C16 and C18 fatty acids, or crop plants mainly storing fructans instead of sucrose in their roots are well-established suitable examples (50). From rape-derived bio-diesel as petrol for cars to highly sophisticated organic chemicals from suitably constructed plant lines, the design of new industrial plants opens wide possibilities for plant biotechnology on a practically unlimited scale.

Initial considerations

A true membrane-bound enzyme will resist extensive washings of membrane preparations with high ionic strength buffers. This is, however, not an absolute criterion. Occasionally, a fraction of a proteinase will remain firmly associated to membranes and the membrane-associated activity will resist repeated washings, despite the absence of a membrane-spanning domain in the primary sequence of the proteinase. For example, 20 of the total activity of thimet oligopeptidase in rat brain homogenates remains tightly associated with membrane fractions (14). The enzyme cannot be removed by repeated washings with 0.32 m sucrose, hypotonic buffers or buffers containing 0.5 m NaCl. However, the enzyme can be completely solubilized by treatment with Triton X-100. A similar situation is seen with the puromycin-sensitive cytosol alanyl aminopeptidase (EC 3.4.11.6). It has been proposed (15) that this enzyme may belong to the class of amphitropic proteins as described by Burn (16). Such proteins...

Bacterial in vitro System of Protein Synthesis and the Cracking of the Genetic Code

Figure 1-17 Sedimentation of 14C uracil pulse-labeled (unstable) RNA of E. coli. The RNA was run on a sucrose gradient for 10 h at 25 000 rpm O.D., optical density 23S, RNA of the large ribosomal subunit 16S, RNA of the small ribosomal subunit 4S, soluble RNA (Ref. 143 , Figure 8). Figure 1-17 Sedimentation of 14C uracil pulse-labeled (unstable) RNA of E. coli. The RNA was run on a sucrose gradient for 10 h at 25 000 rpm O.D., optical density 23S, RNA of the large ribosomal subunit 16S, RNA of the small ribosomal subunit 4S, soluble RNA (Ref. 143 , Figure 8).

The Functional Dissection of Translation

In the context of pursuing ribosomal function, and after the mRNA concept had been established, gentle isolation of messenger-ribosome complexes became a matter of priority in the early 1960s. Particles larger than 70S or 80S appeared on sucrose-gradient patterns and electron microscopic images (see Fig. 1-21 as an

The Structural Dissection of the Ribosome

In view of the complex protein make-up of ribosomes, it is not surprising that RNA was the first ribosomal component to be characterized physically and chemically in terms of sedimentation behavior, molecular weight, and overall base composition. As for the ribosomes, so for rRNA, too, sucrose-gradient centrifugation was crucial. Around 1960, there was still considerable uncertainty about the identity of ribosomal RNA. As long as it was considered to represent the template(s) for protein synthesis, there had been, understandably, no reason to assume that rRNA might be homogenous and well defined. In contrast, credit was given to the idea that ribosomal RNA might be composed of smaller templates that became linked within the particle later on, either non-covalently or covalently. Before RNase-free strains of bacteria became available 221, 222 , the problem of RNA breakdown during preparation could hardly be mastered. Yet the introduction of the separation of RNA from cellular protein...

Glycolysis Converts Carbohydrates into Pyruvate Producing NADH and ATP

Classic glycolytic and fermentative pathways in plants are almost identical with those of animal cells (Figure 11.3). However, plant glycolysis has unique regulatory features, as well as a parallel partial glycolytic pathway in plastids and alternative enzymatic routes for several cytosolic steps. In animals the substrate of glycolysis is glucose and the end product pyruvate. Because sucrose is the major translocated sugar in most plants and is therefore the form of carbon that most nonphotosynthetic tissues import, sucrose (not glucose) can be argued to be the true sugar substrate for plant respiration. The end products of plant glycolysis include another organic acid, malate. In the early steps of glycolysis, sucrose is broken down into the two monosaccharides glucose and fructose which can readily enter the glycolytic pathway. Two pathways for the degradation of sucrose are known in plants, both of which also take part in the unloading of sucrose from the phloem (see Chapter 10)....

Plants Have Alternative Glycolytic Reactions

Sis to sucrose, which is then used to support the growing seedling. In the initial phase of glycolysis, gluconeogenesis overlaps with the pathway for synthesis of sucrose from photosynthetic triose phosphate described in Chapter 8, which is typical for plants. Because the glycolytic reaction catalyzed by ATP-dependent phosphofructokinase is essentially irreversible (see Figure 11.3), an additional enzyme, fructose-1,6-bis-phosphatase, converts fructose-1,6-bisphosphate to fruc-tose-6-phosphate and P during gluconeogenesis. ATP-dependent phosphofructokinase and fructose-1,6-bis-phosphatase represent a major control point of carbon flux through the glycolytic gluconeogenic pathways in both plants and animals, as well as in sucrose synthesis in plants (see Chapter 8). The reaction catalyzed by the PPi-dependent phospho-fructokinase is readily reversible, but it is unlikely to operate in sucrose synthesis (Dennis and Blakely 2000). Like ATP-dependent phosphofructokinase and fructose...

Problems Of Establishment

It is possible to overcome the deficiencies of plant cells at low starting densities by adding small amounts of known chemicals to a medium. For example, Kao and Michayluk (1975) have shown that Vicia hajastana cells or protoplasts can be cultured from very small initial inocula or even from individual cells a standard culture medium was supplemented with growth regulators, several organic acids, additional sugars (apart from sucrose and glucose), and in particular, casein hydrolysate (casamino acids) and coconut milk.

Plant Glycolysis Is Controlled by Its Products

One conceivable benefit of bottom-up control of glyco-lysis is that it permits plants to control net glycolytic flux to pyruvate independently of related metabolic processes such as the Calvin cycle and sucrose-triose phosphate-starch interconversion (Plaxton 1996). Another benefit of this control mechanism is that glycolysis may adjust to the demand for biosynthetic precursors. markedly inhibits the activity of cytosolic fructose-1,6-bis-phosphatase but stimulates the activity of PPi-dependent phosphofructokinase. These observations suggest that fruc-tose-2,6-bisphosphate plays a central role in partitioning flux between ATP-dependent and PPi-dependent pathways of fructose phosphate metabolism at the crossing point between sucrose synthesis and glycolysis.

Oesophageal Adhesion Of Dosage Forms

Tablets are often coated to render them more acceptable to the patient or to protect the drug from gastric acid etc., but the coatings themselves may affect the tendency for formulations to adhere. In vitro studies using isolated oesophageal preparations have concluded that that hard gelatin capsules had the greatest tendency to adhere, followed by film coated tablets, uncoated tablets, with sugar coated tablets demonstrating the least adhesion17 18. It was estimated that hard gelatin capsules have 6 times the tendency to adhere than that of sugar coated tablets and 1.5 times that of soft gelatin capsules when calculated per unit area. The difference between hard and soft gelatin capsules in their tendency to adhere is not borne out in human studies16. Although coated tablets have significantly shorter oesophageal transit times than plain tablets, if they do lodge, they take longer to disintegrate. Coatings made from cellulose acetate phthalate, shellac, methacrylate copolymer and a...

Preribosome Assembly the Proteomic

In the early 1970s, the joint efforts of the Warner and Planta Labs defined the basics of eukaryotic ribosome assembly this remained the core of our understanding for the next 30 years 133, 277, 282, 297, 308, 312, 313 . Metabolic labeling experiments and sucrose-gradient analysis revealed that following transcription, an early 90S pre-ribosome is formed and subsequently partitioned into a 43S and a 66S particle, precursors to the 40S and 60S subunits, respectively (see Fig. 3.2-1). The RNA content of these particles was established as 35S, 27S, and 20S pre-rRNAs for the 90S, 66S, and 43S particles, respectively. The conversion of the 43S particles to 40S sub-units is closely linked to small subunit export. Few RRPs were known at that time and the protein composition of these RNP complexes was not determined.

Differentiated Cells In Callus And Cell Cultures

The level of chlorophyll so far obtained in tissue cultures is well below that found in mesophyll cells of whole plants of the same species, and the rate of chlorophyll formation on exposure of cultured cells to the light is extremely slow compared to the response of etiolated organised tissues. The greening of cultures also tends to be unpredictable and even within individual cells, a range in the degree of chloroplast development is often found. In the carbon dioxide concentrations found in culture vessels, green callus tissue is normally photomixotrophic (i.e. the chloroplasts are able to fix part of the carbon that the cells require) and growth is still partly dependent on the incorporation of sucrose

C to U ApoB RNA Editing csActing Elements

ApoB RNA editing requires the assembly of a sequence-specific higherorder editosome complex that is formed on the RNA. The size of the editosome was identified to be 27S in sucrose gradients using complexes assembled in vitro (63). This equates to a protein complex of about 450 kDa. The minimal enzyme complex that can mediate C to U editing of apoB RNA in vitro comprises two proteins apobec-1, the catalytic subunit, and ACF, a novel RNA-binding protein that serves as the RNA recognition component of the editing enzyme (Fig. 1) (64-66). Other proteins have been identified using detection assays that reflect their ability to interact with apobec-1, ACF, or apoB RNA. These interactions may play a role in modulating the efficiency and specificity of the C to U RNA-editing reaction. Examples of proteins through these approaches include GRY-RBP, CUGBP2, ABBP-1, ABBP-2, AUX240, and hnRNP-C (67-72). Our current understanding of the role of these factors in apoB RNA editing is discussed in the...

Infections Caused By Nonphaemolytic Streptococci

Since many of these streptococci are present in the mouth, upper respiratory tract, genitourinary tract and, to a lesser extent, gastrointestinal tract, they are sometimes involved in pathological processes at these sites, possibly following some local or systemic change in host susceptibility or an alteration in local environmental conditions. A classic example is the manifestation of dental caries that arises following excessive consumption of dietary sugars, particularly sucrose. Alternatively, the streptococci at a mucosal site may gain access to the blood stream because of some local traumatic event and set up an infection at a distant location, such as the heart valve in endocarditis or in the brain or liver, giving rise to an abscess. The key event for infections at distant body sites is bacteraemia.

Phase Changes Can Be Influenced by Nutrients Gibberellins and Other Chemical Signals

The transition at the shoot apex from the juvenile to the adult phase can be affected by transmissible factors from the rest of the plant. In many plants, exposure to low-light conditions prolongs juvenility or causes reversion to juvenility. A major consequence of the low-light regime is a reduction in the supply of carbohydrates to the apex thus carbohydrate supply, especially sucrose, may play a role in the transition between juvenility and maturity. Carbohydrate supply as a source of energy and raw material can affect the size of the apex. For example, in the florist's chrysanthemum (Chrysanthemum morifolium), flower primordia are not initiated until a minimum apex size has been reached.

Pyruvate Enters the Mitochondrion and Is Oxidized via the Citric Acid Cycle

The following two reactions are successive oxidative decarboxylations, each of which produces one NADH and releases one molecule of CO2, yielding a four-carbon molecule, succinyl-CoA. At this point, three molecules of CO2 have been produced for each pyruvate that entered the mitochondrion, or 12 CO2 for each molecule of sucrose oxidized.

Filtering The Transcriptome Enhancing The Value

The Front-end - In order to enhance the signal-to-noise ratio of an experiment, its design can also apply filters at the front-end. For example, rather than interrogating the entire RNA complement of the cell, one can attempt to restrict it only to those transcripts that will be processed into the proteome 49 . We recently carried out such a study in order to explore the efficacy of the front-filter approach 50 . The -agonist, isoproterenol, when administered over a 10-14 day period is a simple and well-characterized protocol that results in a characteristic 20-30 cardiac hypertrophy as measured by the heart to body weight ratios 51 . However, instead of interrogating the entire RNA complements of the treated and untreated animals, only the actively translated RNA was studied. Polysomes derived from the animals' ventricles were loaded on sucrose gradients, size fractionated using velocity density centrifugation and the RNA from these fractions used to select for transcripts that were...

Amide Formation in Senescing Leaves

Asparagine biosynthesis in plants is usually mediated by the action of glutamine-dependent asparagine synthetase (AS), which catalyzes the ATP-dependent transfer of the amide group from glutamine to aspartate, and yields glutamate and asparagine. AS activity has proved difficult to measure because of the presence of natural inhibitors and its instability in vitro (Ireland and Lea, 1999). There is no firm evidence to suggest that AS is located anywhere other than the cytoplasm. Far more information about asparagine synthesis has been obtained by taking a molecular approach. There are three AS genes (ASN1, ASN2 and ASN3) in Arabidopsis thalianta (Lam et al., 1996). The expression of one of these (ASN1) was repressed by light in green tissues and by supply of sucrose in the dark-grown plants. The other two genes were expressed at a lower level than ASN1, and appeared to be regulated in a quite different manner, being induced by light and sucrose, and repressed in the dark. Similar...

Sources for Further Study

A Substance CalledFood. 2d ed. Boston McGraw-Hill, 1989. Presents a variety of perspectives on eating the psychological, the physiological, and the transpersonal. Particularly useful in providing self-help advice and treatment modalities. Examines the compulsiveness of food addiction and sees behavior modification as a means of addressing the addictive behavior.

Transgenic Modification Of Carbohydrate Biosynthesis

Efforts to alter carbohydrate biosynthesis extend back to about 1990 and have proceeded hand in hand with the use of overexpression and antisense inhibition to unravel carbohydrate biosynthesis in plants. The approaches can be divided into those that seek to alter starch quantity through affecting the strength of the carbon source or sink, those that attempt to convert starch to simple sugars, those with the goal of altering the amylose amylopectin Storage organs constitute net consumers or sinks for photosynthetically produced carbon, whereas leaves are the sources. Generally, source-sink balances are regulated by sugar levels (hexoses as well as sucrose) and by stress (74). Willmitzer and his colleagues demonstrated (75,76) that source strength in tobacco is inhibited by accumulation of sugar in the leaves. Expression of a yeast invertase in the cell wall of tobacco cleaved the sucrose normally loaded into the phloem and blocked its export, mimicking a very weak sink. This work was...

Several Subunits of Respiratory Complexes Are Encoded by the Mitochondrial Genome

The maximum yield of cytosolic ATP from the complete oxidation of sucrose to CO2 via aerobic glycolysis and the citric acid cycle The maximum yield of cytosolic ATP from the complete oxidation of sucrose to CO2 via aerobic glycolysis and the citric acid cycle ATP per sucrose0

Ethylendiamine tetraacetic acid

The most important early findings, in a microbiological context, were made by Repaske (1956, 1958), who showed that certain Gram-negative bacteria became sensitive to the enzyme lysozyme in the presence of EDTA in tris buffer and that EDTA alone induced lysis of P. aeruginosa. The importance of tris itself has also been recognized (Leive & Kollin, 1967 Neu, 1969), since it appears to affect the permeability of the wall of various Gram-negative bacteria, as well as the nucleotide pool and RNA, which may be degraded. A lysozyme-tris-EDTA system in the presence of sucrose is a standard technique for producing spheroplasts proto-plasts in Gram-negative bacteria (McQuillen, 1960). During this conversion, several enzymes are

Death Hormone Senescence Signal

Another line of evidence warrants some discussion here, because it has been offered as a test of the senescence hormone hypothesis and implied evidence against the senescence hormone idea (Hamilton and Davies, 1988). 14C-Labeled CO2 was supplied to the pods of pea plants in an effort to determine if labeled metabolites are exported to the shoot apex where the pods cause senescence. There are a number of technical problems with these experiments, but the simplest and most relevant here is isotopic dilution. Given the extent of isotopic dilution (i.e., lowering the specific activity of the 14C-CO2) by the 12C-CO2 produced by the pods or from the atmosphere and then further dilution by metabolites within the source and target tissues, one cannot expect to get detectable levels of 14C into hormone-like molecules which function at very low concentrations (e.g., < 10-6 M). It should also be noted that much of the 14C from 14C-CO2 appeared in sucrose, which as noted above, can promote...

Anthony P Monaco Zoia Larin and Hans Lehrach

YAC libraries have been constructed by preparing and size fractionating high-mol-wt DNA in solution using sucrose gradients (1-3) or in agarose by pulsed field gel electrophoresis (PFGE 4-7). When DNA is prepared in agarose, YAC insert sizes are larger on average for the reason that shear forces seen with DNA in solution are minimized. However, partial degradation of DNA occurs when melting agarose containing high-mol-wt DNA, perhaps owing to metal ion contamination or denaturation (7). The presence of polyamines protects DNA in agarose from degradation at the melting step. By incorporating polyamines in the cloning procedure, mouse and human YAC libraries were constructed with average insert sizes of 700 and 620 kb, respectively (7).

Chemical Modification of Polysaccharides

Catalytic oxidation in the presence of metals and oxygen is claimed as both nonspecific and specific for the 6-hydroxyl oxidation depending on the metals used and the conditions employed for the oxidation. Nonspecific oxidation is achieved with silver or copper and oxygen 196 and with noble metals with bismuth and oxygen 197 . Specificity results with platinum catalyst at pH 6-10 in water in the presence of oxygen 198 . A related patent to produce a water-soluble carboxylated derivative of starch is Hoechst's on the oxidation of ethoxylated starch and another on the oxidation of sucrose to a tricarboxylic acid all the oxidations are specific for primary hydroxyls and use a platinum catalyst at a pH near neutrality in the presence of oxygen 199, 200 .

Specific recommendations for inhibitor use

Figure 1 The protection of trichomonad proteins by leupeptin during electrophoresis. Frozen pellets of Trichomonas vaginalis were lysed by treatment with 0.25 m sucrose containing 0.25 Triton X-100 and proteinase inhibitors, as indicated. The samples were then stored frozen for 2 weeks (S) or used fresh (F). Eiectrophoretlc analysis was carried out by mixing samples with an equal volume of electrophoresis sample buffer 0.0625 m Tris-HCI, pH 6.8, 2 (w v) SOS, 5 (v v) mercaptoethanol, 10 (v V) glycerol and 0.002 bromophenol blue. Some samples were heat-treated by incubation in a boiiing-water bath for 2 mm. Samples were applied to a 7.5 (w v) acrylamlde gel which was run overnight at 5 mA. Proteins were stained with Coomassie Brilliant Blue. None, no inhibitor added AP, plus antipain (0.1 mg ml) LP, plus leupeptin (0.1 mg ml). The positions of standard proteins (molecular sizes in kiiodaltons) are indicated.

Purification of Wild Type Ecotin

The cells are harvested by centrifugation at 6000 g for 30 min, and the supernatant discarded. Osmotic shock is used to selectively lyze the cells and free the periplasm, by resuspending the cell pellet in 10 mM Tris pH 8.0, 25 sucrose plus 15 mg mL1 lysozyme (Sigma) and incubating with shaking for 1 h. The supernatant is retained for the next step where, due to the incredible stability of eco, it can be separated from many of the remaining proteins by acidification. The solution is dropped to pH 2.8 by the gradual addition of 1 M HCl, incubated for 30 min and centrifuged. The acid-soluble supernatant is returned to neutral pH by the addition of 1 M Tris pH 8.0 and followed by the addition of NaCl to 0.3 M. Neutralization is followed by immersion in boiling water for 10 min. After the appearance of visible white protein precipitate, the supernatant is isolated by centrifugation and dialyzed into distilled water. The resulting eco solution is concentrated and purified to homogeneity by...

Short Term Intake Regulation

The secretion of hormones controlling food intake is regulated by the presence of food in the GI tract. The gut is a source of numerous peptides that contribute to the regulation of intake and metabolism. These include, from the small intestine, cholecystokinin (CCK), glucagon-like peptides (GLPs) 1 and 2, bombesin, gastrin-releasing peptide, neuromedin B, glucagon, apolipoprotein A-IV, amylin, somatostatin, enterostatin and peptide YY (3-36) and from the stomach, ghrelin and leptin 7, 11 . Many of the GI hormones and or their receptors are also expressed in the central nervous system, underlining their important role in appetite control 11 . Some enter the central circulation via leaky areas (brainstem and hindbrain) in the blood-brain barrier, or send signals through vagal afferents that are relayed to the hypothalamus. The macronutri-ent-dependent release of gut hormones might explain, at least in part, differences in the satiating and satiety effect of macronutrients. For example,...

Abnormal embryos and plantlets

Differential filtering and sedimentation to separate embryos at different stages of development (Giuliano et al., 1983) can improve the uniformity of embryo populations in suspension cultures. More recently image analysis has been used to select embryos in specific developmental stages (Kurata, 1995). In addition cultures can be maintained on media containing high levels of sucrose (Ammirato and Steward, 1971), and or low levels of abscisic acid (Ammirato, 1973 1974). Both approaches, as well as the addition of imazalil to the culture medium (Werbrouck et al., 2000), limit the number of abnormalities and give a higher degree of synchronisation. High levels of sucrose and abscisic acid induce reversible dormancy in somatic embryos and thus might be used to temporarily suspend growth should this be advantageous in a planned micropropagation programme.

Dietary Components and Intake Regulation

Protein source, in addition to protein quantity, is a determinant of satiety 15 . Greater subjective satiety was reported by young men fed a 50-gram meal of lean fish compared with an equivalent amount of either beef or chicken 21 . Whey and soy protein drinks (45-50 g protein) suppressed food intake 1 h later compared with the energy-free control and with sucrose, whereas egg albumen did not 22 .

The Allosteric Threesite Model as Model Reciprocal Coupling between the A and Esites

Under unfavorable buffer conditions the dissociation rate of a tRNA at the E-site was reported to be 0.3 s-1 33 , whereas the elongation rate, determined under comparable conditions, was reported to be 10 times faster (3 s-1) 34 . This alone suggests that an active mechanism must exist to eject the deacylated tRNA from the E-site. Under near in vivo conditions, the situation is even more significant the dissociation rate of an E-site tRNA from an isolated POST state is much lower and can be measured in hours rather than seconds. POST states can be isolated via overnight centrifugation through sucrose cushions without loss of any deacylated tRNA from the E-site (see, e.g., Ref. 35 ), and native polysomes isolated using a procedure lasting longer than 24 h contain an occupied E-site almost quantitatively 36 . It is clear that the release of a deacylated tRNA from the E-site must be an active process and cannot occur via a simple diffusion process as considered by some authors (see,...

Lifestyle Interventions

Patients with mild to moderate premenstrual symptoms have reported that reducing caffeine, refined sugars, or sodium intake can be helpful. Although increased exercise has been found to reduce symptoms of major depressive disorder, there is no definitive evidence that it results in improvement of PMDD symptoms. There are no recent controlled studies to support the anecdotal reports of the benefits of a healthy diet and exercise for premenstrual syndrome. Nonetheless, these interventions are recommended because of their other benefits and safety.

Nutritional Vitamin and Alternative Complementary Treatment Strategies

Randomized controlled trials ofVitamin B6, calcium, and carbohydrate-rich dietary supplements have demonstrated efficacy in premenstrual syndrome. A recent meta-analysis revealed that vitamin B6 in daily doses of 50-100 mg reduced premenstrual mood and physical symptoms. Calcium supplementation has also been shown to be efficacious in the management of moderate to severe premenstrual symptoms. A randomized controlled trial comparing calcium carbonate to placebo revealed that 1200 mg of calcium carbonate resulted in a significant reduction of physical (water retention, food craving and pain) and emotional (negative affect) premenstrual symptoms.

Results and Discussion

The most commonly used carbon source for tissue culture media is sucrose. The other carbon sources tested for supporting growth include glucose, galactose and also complex carbohydrates such as milkwhey and molasses. Increased sugar concentration favoured synthesis of shikonin in cell cultures of Lithospermum erythrorhizon, diosgenin production in Dioscorea, and anthraquinone in cell cultures of Gallium mollugo. On the contrary, lesser amounts of sucrose favoured the production of ubiquinone 10 in Coleus blumei see 27 . Saccharose as sugar source has shown strongest effect for secondary metabolite content increase in cultures of Catharanthus,

Drug absorption from the colon

Animal models suggest that sugars such as sucrose and glucose, and amino acids, are poorly absorbed from the adult colon63 64, but the relevance to humans has to be demonstrated, since diet has a major effect on colonic physiology and these studies were carried out in the rat and horse. An in vitro study of the permeability of the rat colonic mucosa suggests that the colon excludes molecules on the basis of size and charge65. It has been calculated that the pore size is 2.3A in the human colon compared to 8A in the jejunum and 4A in the ileum66.

Storage Lipids Are Converted into Carbohydrates in Germinating Seeds

Glyoxysome Single Membrane

After germinating, oil-containing seeds metabolize stored triacylglycerols by converting lipids to sucrose. Plants are not able to transport fats from the endosperm to the root and shoot tissues of the germinating seedling, so they must convert stored lipids to a more mobile form of carbon, generally sucrose. This process involves several steps that are located in different cellular compartments oleosomes, gly-oxysomes, mitochondria, and cytosol. Overview Lipids to sucrose. The conversion of lipids to sucrose in oilseeds is triggered by germination and begins with the hydrolysis of triacylglycerols stored in the oil bodies to free fatty acids, followed by oxidation of the fatty acids to produce acetyl-CoA (Figure 11.18). The fatty Malate is transported into the cytosol and oxidized to oxaloacetate, which is converted to phosphoenolpyruvate by the enzyme PEP carboxykinase. The resulting PEP is then metabolized to produce sucrose via the gluconeogenic pathway. acids are oxidized in a...

Fundamental Studies Of Radiation Effects On Biomolecules

Many of the research efforts on amino acids (in particular alanine) and sugars are inspired by their potential dosimetric applications. L-a-alanine is currently used as a reference dosimeter suitable over a wide dose range (McLaughlin 1993) whereas the development of a dosimetric protocol for various sugars (e.g. glucose, fructose, sucrose) plays an important role in the detection of some irradiated foodstuff (Fattibene 1996)._

Fermentation Does Not Liberate All the Energy Available in Each Sugar Molecule

Before we leave the topic of glycolysis, we need to consider the efficiency of fermentation. Efficiency is defined here as the energy conserved as ATP relative to the energy potentially available in a molecule of sucrose. The standard free-energy change (AG0') for the complete oxidation of sucrose is -5760 kJ mol-1 (1380 kcal mol-1). The value of AG0' for the synthesis of ATP is 32 kJ mol-1 (7.7 kcal mol-1). However, under the nonstandard conditions that normally exist in both mammalian and plant cells, the synthesis of ATP requires an input of free energy of approximately 50 kJ mol-1 (12 kcal mol-1). (For a discussion of free energy, see Chapter 2 on the web site.) Given the net synthesis of four molecules of ATP for each sucrose molecule that is converted to ethanol (or lac-tate), the efficiency of anaerobic fermentation is only about 4 . Most of the energy available in sucrose remains in the reduced by-product of fermentation lactate or ethanol. During aerobic respiration, the...

Mechanisms of Ozoneinduced Accelerated Leaf Senescence

Ethylene And Senescence

In recent years, sugars, including glucose and sucrose, have been identified as regulatory signals in a variety of metabolic processes including photosynthesis (Lalonde et al., 1999). Ono and Watanabe (1997) have suggested that when sugar levels increase in the leaf, photosynthetic proteins may decline more rapidly. This may be related to a sugar-induced repression of protein synthesis. There is evidence that when plants are treated with O3, foliar carbohydrates can accumulate (Lux etal., 1997). Fialho and B cker (1996) found transient increases in the levels of several sugars including glucose, fructose and sucrose in leaves of black poplar (Populus nigra cv. Loenen) treated with O3 and SO2 when compared with control tissue. The mechanism by which carbohydrates accumulate in the leaf in response to

Virial Coefficients From Density Measurements

The use of Eq. (21) for the determination of second virial coefficients from partial specific volume measurements 22 is illustrated in Fig. 1, which reappraises results for the nonideality of ribonuclease and chymotrypsinogen that stems from the presence of sucrose 12 , lactose 17 , and glucose 17 . These plots are essentially linear, as Eq. (21) predicts for low enough concentrations and furthermore, there is clearly reasonable agreement between results for the two disaccharides, sucrose and lactose. Second virial coefficients obtained from the slopes, -B.4Af(l - vmPi) (Mm MA), are summarized in Table 1, which also includes values of Mam that emanate from corresponding analyses of results obtained with other polyhydric alcohols (14,15) as the small nonelectrolyte. In general, the dependence of 1 cj> Apo upon molar concentration of small solute in the diffusate, Cm, was essentially linear in the experimental concentration range examined, the slight curvature of the plot for glycerol...

Cell disruption and fractionation

The distribution of cellular proteolytic enzymes among different compartments means that careful disruption of the cells followed by subcellular fractionation can yield samples which are low in endogenous proteolytic activity. Tissues should be disrupted so as to avoid breakage of proteinase-rich vacuoles and lysosomes. This requires the use of gentle techniques (homogenization, freeze-thaw in liquid nitrogen and the omission of detergents such as Triton X-100 which dissolve vacuolar lysosomal membranes). To prevent osmotic rupture of organelles, samples should be homogenized in isotonic solution (e.g. 0.25 m sucrose). Digitonin (0.8 mg ml) can be used to perforate the plasma membrane without the lysis of organelles. After homogenization, the vacuolar fraction can be removed by differential centrifugation. Following a low-speed spin (5 min at 800 g) to remove unbroken cells and nuclei, a further centrifugation for 10 min at 12 000 g in a standard preparative centrifuge should be...

Rat Secretin Receptor

As most- if not all- the G protein coupled receptors, the secretin receptor desensitises rapidly and is internalised through a not yet identified mechanism, insensitive to sucrose but sensitive to Con A 34 . After secretin exposure, the secretin receptor expressed in Cos or CHO cells is rapidly phosphorylated, mainly on Thr residues located essentially in the carboxyl-terminal intracellular tail 35 . Truncation of this part of the receptor does not alter the ligand recognition, nor the coupling to adenylate cyclase but prevents its phosphorylation 36 . However, the truncated receptor remains still internalisable. Thus a dual mechanism limits secretin action a rapid phosphorylation decreases the cyclic AMP production and an internalisation independent of phosphorylation occurs. In transfected HEK 293 cells, it was demonstrated that protein kinases A and C do not markedly affect receptor phosphorylation that is mainly induced by the GRK2 and GRK5 kinases 37 .

Factors affecting the first stage of agglutination

Low-ionic-strength solutions are also used to coat red cells with complement components via the alternative pathway. A few drops of fresh whole blood can be taken into 1-2 mL of 10 sucrose, incubated for 10 min at 37 C and washed, to provide control cells coated with C3 and C4 for the antiglobulin test. For this reason, the use of LISS under uncontrolled conditions can lead to unwanted positive direct and indirect antiglobulin tests.

Materials And Methods

Immature inflorescences of Poaceae and Typhaceae were harvested while unemerged and still covered by leaf sheaths. For Cyperaceae and Juncaceae, inflorescences were gathered while the flowers were small early in their development. The explants were surface sterilized by dilute bleach solution then cultured on primary explant medium DM-8 in the dark at 26-28oC. DM-8 medium 10 contained (in mg l-1, unless indicated otherwise) Murashige and Skoog (MS) salts 11 , Sigma Fine Chemicals) 4,300 Miller's salt solution 6 (w v) KH2PO4 , 3 ml myo-inositol, 100 Vitamix 12 , 2 ml sucrose, 30,000 supplemented with the plant growth regulators adenine hemisulfate, 400 vM picloram, 0.12 indole-3-butyric acid, 1 2,4-dichlorophenoxyacetic acid, 0.5 isopentenyladenine, 0.5 trans-zeatin, 0.5 thidiazuron (TDZ) 3.0 and solidified with Phytagel (Sigma Fine Chemicals), 2000. The secondary medium (0201) contained fewer plant growth regulators at reduced levels 0.2 2,4-D and 0.1 vM TDZ 8 . The tertiary, shoot...

Avoidable Risk Factors

Higher CRC rates in industrialised countries could be related to diets rich in animal products, red meat, animal fats and proteins, and refined sugar, and low in plant-based foods. On the other hand, low-risk diets in developing countries have been observed to be richer in vegetables (particularly cruciferous), High folate diets have been associated with CRC risk reduction 128 . A factor that could increase the chances of developing CRC could be refined sugar 129 , while resistant starches may prevent colon cancer 130 . Omega-3 fatty acids, present in fish oil, could correlate inversely with CRC risk 131 . Chlorinated water could increase the risk of rectal cancers 132 , but the data is not conclusive 133 . Water consumption 134 and methylxanthine-containing beverages such as green tea 135 and coffee may also exert some degree of protection 136 .

Professor Sant Saran Bhojwani

Major research project on Micropropagation of Important Horticultural and Silvicultural species of India was sanctioned to him by the UGC, under which he and his students developed an efficient protocol for clonal propagation of the leguminous tree species, Leucaena leucocephala, and in vitro nodulation of micropropagated plants by Rhizobium to enhance their field survival. He also demonstrated that sugar cubes, produced by Daurala Sugar Mills, was a fair substitute of 'Analar' Grade Sugar used in Plant tissue culture media. The sugar cubes were more than 10 times cheaper than the 'Analar' Grade sucrose. In 1985 the Department of Environment and Forests, Government of India, awarded another major research project to Dr Bhojwani to work on In Vitro Conservation of Endangered Plants. It led to the development of protocols for micropropagation and cold storage of Himalayan Species of three medicinally important plants, viz., Picrorhiza kurroa, Podophyllum hexandrum and Saussurea lappa....

Plants Use Different Strategies to Avoid Salt Injury

As mentioned earlier in the chapter, compatible solutes include glycine betaine, proline, sorbitol, mannitol, pinitol, and sucrose. Specific plant families tend to use one or two of these compounds in preference to others. The amount of carbon used for the synthesis of these organic solutes can be rather large (about 10 of the plant weight). In natural vegetation this diversion of carbon to adjust water potential does not affect survival, but in agricultural crops it can reduce growth and therefore total biomass and harvestable yields.

Acclimation to O2 Deficit Involves Synthesis of Anaerobic Stress Proteins

The mechanism for sensing reduced oxygen levels under hypoxic or anoxic conditions is not completely clear. However, one of the earliest events to occur following lowering of O2 levels is an elevation of the intracellular Ca2+. Evidence suggests that this calcium signal is involved in the signal transduction of anoxia. Within minutes of the onset of anoxia, a rise in cytosolic Ca2+ concentration acts as a signal leading to increases in mRNA levels of alcohol dehydrogenase (ADH) and sucrose synthase in maize cells in culture. Chemicals that block a rise in intracellular Ca2+ concentration also prevent the expression of the genes for ADH and sucrose synthase from being induced by anoxia, and they greatly enhance the sensitivity of maize seedlings to anoxia (Sachs et al. 1996). Further research is needed to resolve these mechanisms and to explain how intracellular Ca2+ concentration signals both the early survival of cells under anoxia and the induction of cell death and aerenchyma...

Methods applications and concepts of metabolite profiling Secondary metabolism

A proposed mechanistic model of the metabolic response ofMedicago truncatula cell suspension cultures to methyl jasmonate elicitation 39 . The data suggest a major reprogramming of metabolism in which as carbon normally destined for sucrose is redirected towards secondary metabolism (triterpene saponin). Figure 1. A proposed mechanistic model of the metabolic response ofMedicago truncatula cell suspension cultures to methyl jasmonate elicitation 39 . The data suggest a major reprogramming of metabolism in which as carbon normally destined for sucrose is redirected towards secondary metabolism (triterpene saponin).

Radical Reactions With Biomolecules

The study of the chemistry of bioradicals is important for the understanding of later biological effects. As we will discuss further damage to the genetic material DNA is the most critical event in radiation exposure of biological systems. Radiation exposure of cells in living organisms may result in cell replication failure or in chromosome aberrations, leading to mutagenesis and carcinogenesis. Radiation damage to proteins, lipids and carbohydrates is relevant for effects such as enzyme inactivation and also has applications in the radiation treatment of foods and drugs where toxicity of radiation products is a point of major concern. Amino acids arid sugars are important for dosimetric applications. The amino acid L-a-alanine is currently used as a reference dosimeter suitable over a wide dose range (Callens 1996 McLaughlin 1993 Van Laere 1993). The study of radiation-induced effects on various sugars (e.g. glucose, fructose, and sucrose) is relevant for the detection of some...

Risks Associated with Pollen and Honeybees

A transgenic plant expressing a PI may affect the bee indirectly via changes in flower attractiveness due to pleiotropic or insertional mutagenesis effects (e.g., nectar volume or sucrose concentration, flower morphology), or directly via ingestion of pollen expressing the inhibitor. Nectar, as a plant secretion containing only a few amino acids,4 is unlikely to be a source of foreign proteins in transgenic plants. Of course the direct effects may be avoided altogether if the PI-gene construct has a promoter which does not allow expression in pollen. However a recent study suggests that promoters may perform differently in different plant species.5 This investigation of the activity in pollen of marker genes controlled by the CaMV 35S and nos promoters showed that both constructs were inactive in Arabidopsis but active, albeit in a highly variable fashion in tobacco.

Answers and Explanations

Both types of transport occur down an electrochemical gradient (downhill), and do not require metabolic energy. Saturability and inhibition by other sugars are characteristic only of carrier-mediated glucose transport thus, facilitated diffusion is saturable and inhibited by galactose, whereas simple diffusion is not. 17. The answer is D III A, B 4 . Solution A contains both sucrose and urea at concentrations of 1 mM, whereas solution B contains only sucrose at a concentration of 1 mM. The calculated osmolality of solution A is 2 mOsm L, and the calculated osmolality of solution B is 1 mOsm L. Therefore, solution A, which has a higher osmolality, is hyperosmotic with respect to solution B. Actually, solutions A and B have the same effective osmotic pressure (i.e., they are isotonic) because the only effective solute is sucrose, which has the same concentration in both solutions. Urea is not an effective solute because its reflection coefficient is zero. 25....

Establishing the Tolerable Upper Intake Level

In the case of added sugars which were also evaluated, no UL was established as there was 'no clear and consistent association between increased intake of added sugars and body mass index' 11 , although there was a trend toward dietary inadequacy with higher intakes of added sugars. In place of a UL, the following statement was provided 'A maximal intake of 25 of energy from added sugars is suggested based on ensuring sufficient intakes of essential micronu-trients that are for the most part present in relatively low amounts in food and beverages that are major sources of added sugars in North American diets' 11 .

Subcellular Fractionation

Further separation is therefore based on other parameters. The use of buoyant density is historically the favored method and allows further purification of contaminated organelles such as mitochondria. In addition, fine-tuning of the density provides a finer separation and allows the separation of microsomes into subcomponents. Various media have been used for this purpose (Rickwood et al., 1983), ranging from sucrose to colloidal silica (Percoll) and to iodinated media. Despite their higher cost, the latter media afford greater flexibility and are now more frequently used (Graham et al., 1983 Wattiaux and Wattiaux-de Coninck, 1983).

Pleiotropic Effects Of Cholesterol Synthesis Inhibition

Experiments with native cell membranes have demonstrated that caveolae and flat rafts can be isolated by biochemical extraction because they resist solubilization by cold nonionic detergents, such as Triton X-100 and Nonidet P-40 (92,125). This process yields aggregated detergent-resistant membranes that sediment at low density in sucrose gradients and are enriched in raft-resident proteins, cholesterol, sphingolipids, and long chain fatty acids (123).

Early Use In Plant Tissue Culture Media

Boric acid reacts with some organic compounds having two adjacent cis-hydroxyl groups (Greenwood, 1973). This includes o-diphenols, hexahydric alcohols such as mannitol and sorbitol (commonly used in plant tissue culture as osmotic agents), and several other sugars, but excludes sucrose which forms only a weak association. Once the element is complexed it appears to be unavailable to plants. This led Lewis (1980) to suggest that because boric acid was required for lignin biosynthesis, vascular plants were led, during evolution, to use sucrose exclusively for the transport of carbohydrate reserves. Although the addition of sugar alcohols and alternative sugars to sucrose can be beneficial during plant tissue culture (see Chapter 4), it is clearly necessary to return to a sucrose-based medium for long-term culture, or boron deficiency may result.

Whole Body Imaging in Oncology

Full Body Tomography

Hydration prior to FDG administration will, as with any tracer cleared by urinary excretion, facilitate tracer clearance from blood pool and the urinary tract. Patients should be encouraged to drink plenty of water, but only water (no sugar containing beverage), prior to FDG administration. After FDG administration

The Digestive System

Digestion of carbohydrates. Starches (e.g. amylose glycogen) are polymers of glucose (C6H12O6). Maltose, lactose and sucrose are disaccharides (Maltose glucose + glucose Lactose glucose + galactose Sucrose glucose + fructose). Absorption of carbohydrates requires that they first be broken down to monosaccharides, namely glucose, galactose, and fructose, which are then absorbed into the small intestine, where they enter the portal blood. This occurs partly through enzymes released by the salivary, intestinal and pancreatic glands. HC1 in the stomach also has a small effect in the hydrolysis of starches.

Overview Of Plant Respiration

Glucose is most commonly cited as the substrate for respiration. However, in a functioning plant cell the reduced carbon is derived from sources such as the disaccharide sucrose, hexose phosphates and triose phosphates from starch degradation and photosynthesis, fructose-containing polymers (fructans), and other sugars, as well as lipids (primarily triacylglycerols), organic acids, and on occasion, proteins (Figure 11.1). Sucrose

Tubers Bulbs and Root Vegetables

Transgenically increasing endogenous cytokinins shortened the dormancy phase in potato (Ooms et al., 1991). Likewise, exogenous application of cytokinins, ethylene and gib-berellins and wounding shortens dormancy in bulbs and tubers, whereas ABA lengthens it (Fig. 22-1 Isenberg et al., 1987). As dormancy ends, the various metabolite reserves, such as starch, in the organs are converted to forms more suited to transport and rapid metabolism in the sprouts, such as sucrose (Stoll and Weichmann, 1987).

Differential Diagnosis

A comprehensive history and physical examination are indicated to rule out other possible causes of the emotional and physical symptoms of PMS. The differential diagnosis includes premenstrual molimina, hypothyroidism, perimenopause and major mood or anxiety disorders. Most ovulatory women experience some physical changes (e.g., breast tenderness, bloating, and food cravings) during the luteal phase. If these physical symptoms do not interfere with normal life functions, the term molimina can be applied. Hypothyroidism can share many of the same symptoms as PMS however there should be no cyclic variation. A thyroid stimulating hormone level is a sufficient screen, if warranted by clinical suspicion. There is also considerable overlap between the symptoms of perimenopause and those of PMS. Many women experience symptoms of emotional irritability, cyclic mastalgia, bloating and hot flashes as part of the perimenopause. It is likely that similar pathophysiologic factors mediate symptoms...

Mitochondria Are Semiautonomous Organelles

The breakdown of sucrose to pyruvate releases less than 25 of the total energy in sucrose the remaining energy is stored in the two molecules of pyruvate. The next two stages of respiration (the citric acid cycle and oxidative phosphorylation i.e., electron transport coupled to ATP synthesis) take place within an organelle enclosed by a double membrane, the mitochondrion (plural mitochondria).

Pharmaceutical Continuous Flow

Groothuis et al. (15) used sucrose, an unmetabolized marker compound that has very low capillary permeability, to initially evaluate the therapeutic feasibility of administering chemotherapy by the intraventricu-lar route. Sucrose was infused by osmotic minipump into the lateral ventricle of a rat for 7 days, yielding the concentration profile exhibited in Figure 9.5A, a profile well fit by theoretical Equation 9.5 using published diffusion and permeation constants for sucrose (16). In this experiment, the penetration distances to one-half and one-tenth the surface concentration were 0.9 and 3 mm, respectively. In a subsequent study, Groothuis et al. (17) continuously infused ARA-C into the ventricles of rat brain over 7 days. They found that even with continuous administration of ARA-C, tissue concentrations dropped to one-half the surface concentration at a penetration distance of 0.4 mm and to about one-tenth the surface concentration at a penetration distance of 1.0 mm (Figure...

The Electron Transport Chain Catalyzes a Flow of Electrons from NADH to O2

For each molecule of sucrose oxidized through glycolysis and the citric acid cycle pathways, 4 molecules of NADH are generated in the cytosol and 16 molecules of NADH plus 4 molecules of FADH2 (associated with succinate dehydrogenase) are generated in the mitochondrial matrix. These reduced compounds must be reoxidized or the entire respiratory process will come to a halt.

Exchange Membrane Chromatography

HIV-1 based lentiviral vectors could be directly captured and eluted from clarified cell culture supernatants on a 60 ml membrane bed volume Mustang Q strong anion exchange membrane chromatography capsule without any pre-concentration step. The vector was purified some 1000-fold as determined by a p24 ELISA assay with a 30 recovery of infectious particles. The purity and recovery of the vectors was similar to those purified using a size exclusion chromatography step. This latter method was used to purify the LV vectors that are currently being used in clinical trials 71 . The low recovery of infectious particles could be due to a combination of factors including variability in the infectivity assay and the presence of high salt in elution buffer that may compromise the integrity of the lentivirus envelope that is responsible for cellular attachment and fusion. In order to improve the infectious particle recovery during chromatographic purification of enveloped virus vectors, it may be...

From Microsomes to Ribosomes

And chemical composition, became linked on experimental grounds to protein synthesis in vivo 97-102 and in vitro 70, 96, 102, 103 . Around this time, hints were also accumulating that eukaryotic microsomal material was quite heterogenous in size as well as in composition. It took another decade before the the isolation of active cyto-plasmic particles through sucrose-gradient centrifugtion became a laboratory standard. To obtain 'purified' microsomes became one of the major issues in the development of cell-free protein synthesis around 1955 104 . reversibly into a 50S and a 30S component 116, 117 . Gradually, in a decade of painstaking isolation attempts, in which sucrose-gradient centrifugation came to occupy a central place, the confusion about the size of the RNP particles cleared up, and it was realized that the secret of stabilization lay chiefly in the concentration of divalent Mg2+ ions. Work on a variety of particles from other sources began to converge on two distinguishing...

Fraction Number

Sucrose gradient of lysed reticulocytes after incubation with 3H leucine. After lysis and low-speed centrifugation, the ribosomes were pelleted three times at 28 000 rpm and resuspended with a homogenizer. A 5-20 sucrose gradient was used. The numbers next to the arrows represent the sedimentation constants associated wih each peak (Ref. 214 , Figure 3).

Lipid breakdown

From a chemical standpoint, plant respiration can be expressed as the oxidation of the 12-carbon molecule sucrose and the reduction of 12 molecules of O2 This reaction is the reversal of the photosynthetic process it represents a coupled redox reaction in which sucrose is completely oxidized to CO2 while oxygen serves as the ultimate electron acceptor, being reduced to water. The standard free-energy decrease for the reaction as written is 5760 kJ (1380 kcal) per mole (342 g) of sucrose oxidized. The controlled release of this free energy, along with its coupling to the synthesis of ATP, is the primary, though by no means only, role of respiratory metabolism. To prevent damage (incineration) of cellular structures, the cell mobilizes the large amount of free energy released in the oxidation of sucrose in a series of step-by-step reactions. These reactions can be grouped into four major processes Glycolysis involves a series of reactions carried out by a group of soluble enzymes...

Symplectin

The soluble form of the Symplectoteuthis photoprotein was isolated and purified from the granular light organs of the squid and was named symplectin (Takahashi and Isobe 1994 Fujii et al 2002). The light organs were first extracted with a pH 6 buffer containing 0.4 M KCl to remove impurities, and then symplectin was extracted from the residue with a pH 6 buffer containing 0.6 M KCl. All solutions used in the experiments contained 0.25 M sucrose, 1 mM dithiothreitol, and 1 mM EDTA. The 0.6 M KCl extract was chromatographed by size-exclusion HPLC on a TSK G3000SW column. Symplectin was eluted as two major components of oligomers, having molecular masses of200 kDa or more, and a minor component of monomer (60 kDa). All processes of extraction and purification were carried out at 4 C. Warming up a solution of symplectin, adjusted to pH 8, to room temperature causes the luminescence reaction to begin, and the light emission lasts for hours.

Enzyme Preparations

Microsomes are prepared from tissues by first suspending 2-5 g of tissue in 4 volumes of 0.25 M sucrose containing protease inhibitors (1 mM phenylmethylsulfonylfluoride, 10 mM EDTA, 1 mM N-ethyl maleimide, 0.5 mg mL leupeptin and 0.1 mg mL pepstatin) in case there might be slight protease modifications of the enzymes (see Note 10), and homogenizing 3 to 5 times with a Polytron at the maximum speed for 1 min each. The homogenate is centrifuged at 12,000g (occasionally 10,000g or 20,000g) in a fixed-angle rotor for 20 min, and the supernatant fluid is then centrifuged at 105,000g for 1 h. The pellets are suspended in 0.25 M sucrose and recentrifuged at 105,000g for 1 h to wash the resulting microsomal fraction (see Note 11). Greatest experience has been with microsomal preparations from chick embryo epiphyseal cartilage (see Note 12) and mouse mastocytomas. 3. Fractionation of chick embryo epiphyseal cartilage microsomes to provide Golgi is performed by a sucrose density-gradient...

Secretion rate

Salivary amylase acts on food starch while the acidity of the mixture is low, around pH 6, but ceases when the acidity of the mixture increases with greater acid secretion. Gastric pepsins account for only about 10 to 15 percent of the digestion of protein and are most active in the first hour of digestion. The stomach is primarily a processing organ and not an absorptive one and is not essential to life. It is possible, for example, after total gastrectomy (the complete removal of the stomach) for a person to remain, or to become, obese because most of the digestion and absorption of food takes place in the intestine. The stomach can absorb some substances, including glucose and other simple sugars, amino acids, and some fat-soluble substances. A number of alcohols, including ethanol, are readily

Carbohydrates

The principal dietary carbohydrates are starches, sucrose and lactose. Starch is a glucose-containing polysaccharide with a molecular weight which varies from 100,000 to more than 1 million. The two major polysaccharides of starch are amylose and amylopectin. Indigestible carbohydrates e.g. cellulose are the main constituents of dietary fibre.

Starch Deposition

Starch biosynthesis with its key enzymes and metabolites is diagrammed in Fig. 2. Photosynthate is generally supplied as sucrose via the phloem of the maternal tissues. Both source and sink strength are critical to starch yield in storage organs. In some plants, breakdown and resynthesis of sucrose appear necessary to maintain a sucrose gradient and thus sink strength, although this is not the case in others such as barley. The sucrose taken into the endosperm is subsequently converted into UDPglucose by sucrose synthase EC 2.4.1.13) sucrose + UDP > UDPglucose + D-fructose This is a reversible reaction but, under the conditions found in storage tissues, the breakdown of sucrose is favored. In many plants, sucrose synthase activity appears to be important to overall sink strength and hence yield. Antisense-mediated reductions in sucrose synthase levels in transgenic tomato (24) and potato (25) reduce overall starch biosynthesis, as do mutations to the sucrose synthase genes such as...

Conclusions

Plants are well suited as producers of modified starch and novel carbohydrates. Photosynthesis supplies a sucrose feedstock to carbohydrate storage organs, and the many intermediate steps in the conversion of sucrose to starch represent potential branch points at which the sugar may be shunted to new products. Storage starch, being insoluble, is not physiologically active, and the many mutations affecting starch synthesis demonstrate that a wide variety of structures can be tolerated by the plant. Tubers and storage roots (beets) are not required for propagation, so the storage starch in these organs need not be accessible to the plant for turnover. Cereal grain starch is, however, important in germination therefore, modification or replacement of this carbon source must take physiological needs into account.

Graft Polymers

Other grafts to natural materials are exemplified by Dordick's work 173 in which he produced polyesters from sugars and polycarboxylates by enzyme catalysis of the condensation polymerization. These polymers and the method of synthesis may well be the future of renewable resource chemistry.

Metabolic Changes

During senescence, daylily flowers lose up to 95 of their sugars and 14C-sucrose applied to attached senescing flowers is rapidly translocated to other parts of the plant, particularly to the developing flower buds (Bieleski, 1993). In gladiolus, removal of this carbohydrate source by removal of the mature flowers, caused a substantial reduction in the opening of subsequent flowers (Waithaka etal., 2000), which is consistent with the demonstrated export of radioactively labeled sugars from wilting gladiolus florets to younger buds (Yamane et al., 1995).

Flotation Techniques

The specific gravity of helminth ova and larvae and many protozoan cysts is in the range 1.051.14 and thus, if a liquid medium that is denser than the parasites is utilized, they will rise to the surface. They can then be skimmed from the surface using a microscope cover slip and examined. Several flotation media have been described, including concentrated aqueous sodium chloride solution (specific gravity 1.121.2), sucrose solutions (specific gravity 1.180) and 33 zinc sulphate. In general, these techniques are rather difficult to perform and if the eggs are left in the hypertonic solutions for a prolonged period, the morphology may be significantly altered, making identification difficult (Denham and Suswillo, 1995).

Phosphate

Although the concentration of phosphate introduced into plant culture media has been as high as 19.8 mM, the average level is 1.7 mM and most media contain about 1.3 mM. However many reports indicate that such typical levels may be too low for some purposes. When phosphate is depleted from MS medium, there is an increase in free amino acids in Catharanthus roseus cells, because protein synthesis has ceased and degradation of proteins is occurring (Ukaji and Ashihara, 1987). Phosphate (starting concentration 2.64 mM) and sucrose were the only nutrients completely depleted in Catharanthus roseus batch suspension cultures, and the period of growth could be prolonged by increasing the levels of both (MacCarthy et al., 1980). MS medium contains only 1.25 mM phosphate which may be insufficient for suspension cultures of some plants. The phosphate in MS medium is insufficient for Cardamine pratensis suspension cultures, all having been absorbed in 5 days it is however adequate for Silene...

Subunit

Results in accumulation of Ub conjugates.44 Wilkinson et al have reported that S2 interacts genetically and biochemically with S4.96 Biochemical studies in our laboratory confirm this result (Gorbea, Taillandier, and Rechsteiner, in preparation). We translated S2 in rabbit reticulocyte lysate in the presence of 35S -methionine and used the radiolabeled protein to probe RC subunits separated by SDS-PAGE and transferred to nitrocellulose. 35S-labeled S2 bound S4 present on the membrane, but 35S -S4 did not bind to S2 in the reciprocal assay. Therefore, we cotranslated both subunits in lysate and sedimented the radiolabeled proteins on sucrose gradients. S2 and S4 form a dimer that sediments near the aldolase marker (158 kDa), and both proteins can be immunoprecipitated with either anti-S2 or anti-S4 polyclonal antibodies. In summary, there is good evidence indicating that the non-ATPase S2 binds S4, an ATPase, within the regulatory complex.

Explant

Isolated plant roots can usually be cultured on relatively simple media such as White (1954) containing 2 sucrose. Liquid media are preferable, as growth in or on a solid medium is slower. This is presumably because salts are less readily available to the roots from a solidified medium and oxygen availability may be restricted. Although roots will accept a mixed nitrate ammonium source, they will not usually grow on ammonium nitrogen alone. Species, and even varieties or strains, of plants, are found to differ in their requirement for growth regulators, particularly for auxins, in the root culture medium.

Media Composition

The basic media composition for microspore culture protocols in various oilseed brassicas have mostly been the same over the years. Initial reports of anther culture in oilseed brassicas emphasized the role of increased sucrose concentration in culture media 51, 60 . Subsequent to this, it was established that high sucrose concentration (10 to 13 ) is essential for microspore embryogenesis in anther as well as isolated microspore culture in B. napus 49, 52, 56 . Keller et al. 60 demonstrated that L-serine was an important constituent of the anther culture media in B. napus. Later, Lichter 52 demonstrated that basal Nitsch and Nitsch 100 medium supplemented with glutamine, glutathione and L-serine was most congenial for anther culture in the same species. Off late Nitsch and Nitsch medium 100 modified by Lichter 52 , commonly known as NLN medium, has become the most frequently used media for isolated microspore culture in oilseed brassicas.

Twitching Motility

That were found to repress twitching motility were generally those that increase the osmolarity of the medium such as NaCl (300 mM), KCl (300 mM), sucrose (5 w vol), glucose (10 w vol), glycerol (5 vol vol), and polyvynilpyrroli-done (2 w vol). On the other hand, we found that increasing the tryptone and yeast extract components of Luria Bertani medium either together or individually to 3-5 times standard concentrations was stimulatory to colony expansion (C.B. Whitchurch, A.B.T Semmler, and J.S Mattick, unpublished observations). A similar result was obtained when tryptone (3-5 w vol) was added to the base media. Interestingly, addition of equivalent amounts of Casamino acids to the base media had the opposite effect on colony expansion and was inhibitory (C. B. Whitchurch and J. S. Mattick, unpublished observations). Both tryptone and Casamino acids are derived from casein, the former being a tryptic digest of the protein and thus comprised of peptides, and the latter an acid digest...

Pain and Discomfort

Guidelines for the management of pain in newborns have been published (AAP, 2001 Ped 108 793 Anand et al., 2002 Arch Pediatr Adolesc Med 155 173). The most widely used medications for the treatment of severe acute pain are morphine and fentanyl. Soothing measures (nonnutritive sucking of oral sucrose on a nipple) are also provided during minor procedures (Stevens and Ohlsson, 2000 Stevens et al., 2004). Studies have not consistently demonstrated the benefits of the routine treatment of mechanically ventilated newborns with narcotics (preemptive analgesia), however (Bellu et al., 2005 Cochrane 7 CD004212 Grunau et al., 2006).

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