Cyclical Ketogenic Diets Review

Keto Resource

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Keto Resource Summary


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Dietary Guidance For Carbohydrates

Dietary guidance for consumption of carbohydrates has resembled laboratory analysis of carbohydrates take away the fat and protein and the remainder must to be carbohydrate. Nutritionists generally accept the fact that humans don't need more than 10-12 of kilocalories from protein, and less than 30 of kilocalories from fat. Subsequently, intake of carbohydrate should be 55 of kilocalories or more. Human diets historically have contained 40-80 of their energy as carbohydrate (5), although as income increases, so does the fat content of the diet while the carbohydrate content of the diet, especially the starch, decreases. carbohydrates and starch. Sugar recommendations are often divided into naturally recommended Americans increase the consumption of complex carbohydrates and naturally occurring sugars from about 28 of energy intake to 48 of energy intake. Such a recommendation would be difficult to follow with the limited information on the complex carbohydrate content of foods....

Dietary Guidance For Complex Carbohydrates

In the U.S. the term complex carbohydrates was used in the McGovern report and included digestible carbohydrates or starch (2). The British Nutrition Foundation Report (13) included both starch and non-starch polysaccharides as complex carbohydrates. The British report defined polysaccharides as carbohydrates with 20 or more sugar residues. The British report stated that there are a variety of foods which are rich in complex carbohydrates and display great diversity in their physical and biological properties. Thus, it is difficult to make generalizations about properties of complex carbohydrates. They also noted that there is more overlap between the behavior and properties of the starches and non-starch polysaccha-rides than was previously thought. Resistant starch is probably metabolized similar to some dietary fiber components in the gut. Thus, it appears that complex carbohydrate should contain both dietary fiber and starches and we must recognize that substances (such as...

Suggested Alternatives to the Term Complex Carbohydrates

Refer to the benefits of diets high in complex carbohydrates, defined essentially t carbohydrates is permitted and has been taken to mean only starch which is the carbohydrates has been taken to mean either starches or starches plus dietary fiber. Thus, the term is a source of confusion, is difficult to define in a manner that can be verified analytically and has little scientific meaning for many scientists and lay people alike (1, 2). There is a need to simplify and more clearly define in chemical terms what is meant by the advice to consume foods high in complex carbohydrates. It is time to re-think the use of this chemically undefined

Measurement And Identification Of Carbohydrates

As the physiological effects of different polysaccharides become better understood it is appropriate to bring improved definition and precision to the area of food carbohydrates. This message is important when one considers that as recently as 1992 carbohydrate was still measured by difference in USDA Handbook 8, a practice subject to error because it includes minor non-carbohydrate components and is subject to cumulative analytical error derived from the analyses for protein, fat, moisture and ash. The FDA does not permit the term complex carbohydrate but requires the term other carbohydrate which is defined as the amount of carbohydrates remaining after subtraction of dietary fiber, sugars and sugar alcohols from total carbohydrates (3). This introduces another ill-defined and confusing term and continues to avoid mentioning by name the chemically well-defined term starch.

Defining Simple And Complex Carbohydrates

Carbohydrate-rich foods contain mixtures of sugars, starches, and fibers. Different criteria are used to define the analytic and physiologic distinctiveness of these components. Because the impact of carbohydrates on human nutrition is more rigorous than inherent in simplified definitions, integrated analytic and nutrition terminologies are necessary to convey the importance of carbohydrates in human health. Although there is an established analytic nomenclature for carbohydrates, appropriate descriptors are required to indicate their collective physiologic attributes and roles. The term complex carbohydrates was adopted to simplify the dietary contributions of a broad and chemically diverse group of carbohydrates. For instance, variable inclusion or exclusion of total dietary fiber is one of the recognized incongruities related to the common use of the term. The lack of accepted measurements for defining the components of complex carbohydrates resulted in the exclusion of information...

Summary Of Aoac Survey On Complex Carbohydrates

In 1992 the AOAC International Task Force on Nutrients Labeling Analysis and its Subcommittee on Carbohydrates Methodology concluded that more work was necessary to develop an analytic methodology for complex carbohydrates. In 1993 AOAC International subsequently established refereeship in the area of complex carbohydrates to accommodate the analytic needs. As the first step in establishing analytic methodology an international survey on complex carbohydrate definition and analysis under the auspices of AOAC International was initiated. The survey was designed to determine if a consensus could be reached on the definition of complex carbohydrates and associated methodologies. Approximately 200 survey questionnaires were distributed to professionals in the area of carbohydrates and 114 replies were received. The survey results indicated that complex carbohydrates should include available starch and total dietary fiber. In the previous two international surveys on dietary fiber...

Covalent Attachment Of Biotin To Carbohydrates

Biotinylation of protein carbohydrates requires oxidation of vicinal carbohydrate hy-droxyl groups to aldehydes that can bind biotin-LC-hydrazide. Sodium m-periodate is used as the oxidizing agent for glycoproteins in solution, but enzyme-mediated oxidations can be used to oxidize glycoproteins of cells or membrane fractions. Briefly, glycoproteins are oxidized, excess oxidizing agent is removed by gel-filtration, biotin reagent is added, and the biotinylated glycoprotein is purified in a microconcentrator.

International Survey on Complex Carbohydrates Definition

Respondents were first asked to select from a provided list of carbohydrate components those which, in their view, should be included in the definition of complex carbohydrates for food labeling purposes. The question was presented in a multiple choice format, with the possibility of writing in additional options. The various options are listed in Table 2. The 107 respondents were also asked to explain and justify their choices. Most respondents felt that available starches (72 ) should be included in the definition of complex carbohydrates. The majority of respondents also listed the terms non-starch polysaccharides (69 ), total starches (58 ), and unavailable or resistant starches (58 ) to be included in the complex carbohydrate fraction. Approximately one-third of respondents included both available (33 ) and un- available oligosaccharides (32 ) and lignin (29 ) in their definition of complex carbohydrates. Further data analyses showed that 62 of respondents indicated that both...

Use Of Hpaepad For Determination Of Complex Carbohydrates And Dietary Fiber

The increasing utilization of HPAE-PAD for determination of complex carbohydrates and dietary fiber constituents is illustrated by the number of publications on these topics in the last few years (11,14-23). Englyst et al. (11,14,15) have used HPAE-PAD to determine neutral sugars, uronic acids, and hexosamines from non-starch polysaccharides. Corradini et al., have developed methods for the determination of nutritionally significant carbohydrates (16). Jane et al. have applied the technique to study debranched amylo-pectins (17). Coussement et al. have developed methodology for the determination of inulin and oligofructose in food products (18). Koizumi et al. have studied chain length distribution of amylopectins and oligo- and polysaccharides with DP 50 (19,20). Fahey et al. have used HPAE-PAD to determine the neutral monosaccharide composition of various fibrous substances and have compared the HPAE-PAD method with the standard colorimetric procedure for the determination of pectic...

High Performance Anion Exchange Chromatography Of Carbohydrates

In order for pulsed amperometry to have utility for the chromatographic determination of carbohydrates, a high resolution separation method is required that is also compatible with high pH. This requirement has been met by the development of column technology which allows high resolution separations of carbohydrates by anion exchange chromatography at high pH. Carbohydrates are often thought of as neutral compounds, but in actuality they are weak acids with pKa's in the range of 12-14 (Table 1). At these pH levels, oxyanion formation occurs (Figure 6) allowing separation by ion exchange. Ion exchange separations are based on the relative affinity of the analyte ion in competition with the eluent ion for the same exchange sites (Figure 7). The greater the affinity of the ion, the longer the retention time. For carbohydrates, subtle structural differences and small differences in pKa can cause significant differences in retention characteristics. Table 1 Dissociation constants of some...

Determining The Stoichiometry Of A Proteinprotein Complex Containing Carbohydrates

If a protein complex contains carbohydrates, the three-detector method is required to calculate its polypeptide molecular weight and determine its stoichiometry. In this section, we will focus on how to determine the stoichiometry of such a complex (Wen et al., 1996a,b).


A twofold increase in carbohydrate intake will result in a considerable decrease in lipid and protein intake (see Figure 12.3d). The adverse effects after excessively high carbohydrate intake are attributed to decreased intakes of the other macronutrients, rather than to the toxicity of carbohydrates. A high dietary intake of specific carbohydrates has been reported to affect the health of small groups of the population. The absence of disaccharidases in the brush border of the intestinal mucosa connected with genetic as well as contracted disorders gives rise to absorption disturbances and chronic diarrhea. Deficiencies of the disaccharidases sucrase and maltase are rare. On the other hand, lactase deficiency occurs rather frequently. Symptoms of lactose intolerance are usually mild or absent unless large quantities are taken, e.g., a liter of milk, which contains 50 g of lactose. The cause of lactase deficiency may be of three types. First, there is the rare congenital lactase...

Dietary Guidance For Dietary Fiber

High-carbohydrate, low-fat diets containing 20 to 35 grams dietary fiber per day g note that this recommendation is not appropriate for the pediatric or geriatric populations. Food, not dietary fiber supplements, provides the best means of increasing daily consumption of both soluble and insoluble fiber. The practice of taking fiber supplements to replace consumption of high-carbohydrate, nutrient-dense foods is not supported at this time by any health care authorities as a component of a balanced diet. In 1991 the United Kingdom issued recommendations for dietary fiber that state that . . . diets should contain an average of 18 g day (range of 12-24) non-starch polysaccharide (NSP) from a variety of foods whose constituents contain it as a naturally integrated component (11).

Prenutrition Labeling And Education Act Of 1990

Even before the issuance of the first Dietary Guidelines for Americans in 1980, the FDA and the food industry had realized the problems created by there being no indication on the food label of the complex carbohydrate content of a food. At the same time dietary guidance almost universally advised consumers to increase their consumption of complex carbohydrates . For example, in 1978, the Kellogg Company had submitted a petition to the FDA to voluntarily permit the declaration of (1) grams per serving of starch and related carbohydrates , (2) grams per serving of sucrose and other sugars , and (3) grams per serving of total carbohydrates the sum of (1) and (2) . The Agency did not grant the Kellogg petition, but in the years from 1980 to 1993 many cereal companies listed either complex carbohydrates or starches and related carbohydrates on the labels of their products and the FDA took no regulatory actions. In 1979 the FDA in cooperation with the United States Department of...

Postnutrition Labeling And Education Act Of 1990

When Congress passed the Nutrition Labeling and Education Act of 1990 (NLEA) one of the principal differences of that act compared to the FDA's 1990 proposal was the inclusion of complex carbohydrates as one of the components that was mandatory on the nutrition label (Section 403(q)(1)(D)) (11). In November 1991 the FDA published a revised proposal for mandatory nutrition labeling to conform agency regulations to the NLEA requirements. (12) Following the lead of Congress the agency proposed that complex carbohydrates be made mandatory on nutrition labels in that document. The definition (starches and dextrins with 10 or more saccharide units) remained as had been proposed in July 1990. However, the November 1991 re-proposal recognized that the NLEA gave the FDA some latitude in determining which nutrients should be listed on the nutrition label. Section 403(q)(2)(B) of the act allows the Secretary (and, by delegation, the FDA) to determine whether information relating to nutrients...

Chicory Fructooligosaccharides as Prebiotics

Carbohydrates *Bulking index g of increase in fresh fecal weight g of carbohydrates eaten ( ) Indicates either added to rat diet or g eaten per day *Bulking index g of increase in fresh fecal weight g of carbohydrates eaten ( ) Indicates either added to rat diet or g eaten per day

Physiological Effects And Labeling

Because high carbohydrate diets containing foods high in starches and dietary fiber can have beneficial health effects, the most useful and practical position at present for the purposes of labeling is to identify starches and dietary fiber separately. This approach to simplifying the definitions, analytical and labeling issues would facilitate healthy food choices by the consumer.

Methodology Development

The McGovern commission recommended that FDA establish a definition of complex carbohydrates. The FDA methodology for measuring complex carbohydrates is determined by summing the total dietary fiber and available starch measured in the sample. Using 80 aqueous ethanol simple carbohydrates (sugars In a recent survey conducted by the AOAC International Food and Nutrition Committee the FDA methodology appears to meet the definition criteria most frequently cited as appropriate by scientists working in the area of complex carbohydrates. However, a number of methodology issues still require resolution. Two basic issues concern completeness of starch digestion by the alpha-amylase-protease-amyloglucosidase mixture, and elimination of the long and tedious step of filtrate evaporation through implementation of newer, more sensitive HPLC detectors. Complex carbohydrate methodology must address quantification of oli-gosaccharides resistant to digestion in the human alimentary system. Finally,...

Summary And Conclusion

This conference resulted in a foundation upon which consensus will be built in the future. Further, it provided a forum for the articulation of the critical need for carbohydrates in human nutrition. We have come a long way in the past 25 years in the food and nutrition sciences and this conference well illustrated that point. At the White House Conference on Nutrition in 1969, macronutrients were never even mentioned as being necessary to nutrition only micronutrients were emphasized. In 1977 the term complex carbohydrate was used without definition in the Dietary Goals for the United States (1). This conference encapsulated the tremendous strides that have been made concerning the chemical, nutritional, biologic, and physiologic importance of carbohydrates in health and disease. With this level of apparent clarity it is difficult at first to identify a problem for the food label. After all, if consumer health would benefit why not simply do it However, a problem does exist in that...

Epidemiologic Studies

Epidemiologic studies performed by Burkitt, Trowell, and Walker indicate there are great differences in intakes of dietary fiber depending on geographical location, and simultaneously significant negative correlation between the consumption of dietary fiber and the prevalence of CHD (1, 2). Nutritional studies in which diet composition of people living in rural areas in Africa and in urban areas in Europe were compared indicate that diets rich in fiber and complex carbohydrates may prevent lipid metabolism disorders and atherosclerosis (3, 4). This hypothesis was further confirmed by the observation that Africans who adopt European lifestyles and diets based on refined products showed a significant increase in the prevalence of hyperlipidemias and CHD.

What Is Dietary Fiber

Tant oligosaccharides, defined as oligosaccharides that are resistant to hydrolysis by human alimentary enzymes, can be used synonymously with the term ''unavailable oligosaccharides.'' This proposal was adopted at the AOAC Workshop I on Complex Carbohydrates held in Nashville, TN, USA in October, 1995. 2

The Physiologic Effects

Complex carbohydrates are important sources of dietary energy, and those that resist digestion are now emerging as important components of the diet. Some of these carbohydrates are fermented like soluble dietary fiber. Others resist fermentation and are excreted in the same manner as insoluble dietary fiber.

The 1995 AOAC International Workshop Results

To conclude the results of the three international surveys, an AOAC International Workshop was also held in Nashville, Tennessee, on September 15-16, 1995, with 57 participants from the United States, Canada, and Europe. The foremost authorities in the world from industry, academia, and government participated. The Workshop is a follow-up to the international surveys as well as the activities of the Carbohydrates Subcommittee of the Nutrients Labeling Analysis Task Force of 1992-1993 and the International Life Science Institute (ILSI)-North American Workshop held in November 1994 in Washington, D.C. (14). The workshop results indicated that there is a general agreement among industry that complex carbohydrates equal available starch and dietary fiber (15). There was a general agreement among the workshop participants that dietary fiber should be included in the definition of complex carbohydrates and that ROs are part of dietary fiber. Twenty-nine persons, representing 85 of the 34...

Complex Carbohydrate Definition

The term ''complex carbohydrates'' was proposed as a mandatory item on the U.S. food nutrition label in 1991. However, the final regulations in 1993 do not allow the use of the term on the nutrition label because of a lack of clear definition and methodology. At the end of 1994, ILSI North America's Technical Committee on Carbohydrates sponsored a workshop entitled ''Complex Carbohydrates The Science and the Label.'' The following resulted 1. Complex carbohydrates is a familiar term to consumers who are aware of the need to increase their consumption. However, consumers are unable to readily determine this information from the nutrition label. 2. Based on a recent AOAC International survey, complex carbohydrates definition includes both DF and digestible complex carbohydrates (e.g., starch). Since RO is generally considered part of DF, it is also a complex carbohydrate. 3. An AOAC assay for complex carbohydrates under development will measure both DF and digestible complex...

Fermentation to Short Chain Fatty Acids SCFA

Studies in rats and man have shown that polydextrose is partially fermented in the large intestine (18-21, 32). Because not all fibers are degraded equally by bacteria, a range of 0 (nonfermentable, insoluble) to 4.0 (fermentable, soluble) kcal g have been used for these fibers unless proven otherwise. As previously described, polydextrose has been determined to be 1 kcal g. Many fibers can be fermented by bacteria in the large intestine to produce hydrogen, methane, carbon dioxide and SCFA (also referred to as volatile fatty acids, VFA). The SCFA are rapidly absorbed from the gastrointestinal tract through the hepatic portal vein and contribute to the energy balance of the body. These contributions include inhibition of hepatic cholesterol synthesis by propionate (33) and aptosis of cancer cells by butyrate (34,35). Qualitative measurement of SCFA was conducted in PDX studies (7) and revealed typical profiles of acetic, butyric and propionic acids. Of the total radioactivity...

Gastrointestinal Function

Changes in gastrointestinal function are most pronounced for insoluble fibers. However, Oku et al. (38) found that polydextrose fed to rats at a dose of 3 of the diet partially demonstrated the DF actions of increased fecal volume and weight, decreased transit time and increased moisture of fecal samples. Tomlin and Read (39) found that polydextrose increased fecal mass and softened stools. Nakagawa et al. (40) found that polydextrose ingestion led to softer stools. Also, Wang and Gibson (36) measured growth of colonic bacteria in batch fermenters with various carbohydrates added. Over 12 h, polydextrose caused an increase in total bacteria similar to the other carbohydrates (pectin, starch, inulin, oligofructose, fructose). This is important because bacteria form a significant portion of fecal bulk.

Definition Of Dietary Fiber

The above examples show that we need to take into account the botanical structure of the cell walls in our foods This structure plays an essential role in the physiological effects of dietary fiber. At this time, the notion of integrity of the botanical structure is not accounted for in the term ''complex carbohydrates'' and also, by present definitions, the term ''dietary fiber'' fails to fully acknowledge its importance. Because analysis of dietary fiber tells nothing about the origin of a sample or the integrity of its structure, clearly the methods alone cannot be used to define dietary fiber. It appears that the way to take structural integrity into account is to refer to it specifically in the definition of dietary fiber. Material complying with conditions 1 and 2 above would be adequately measured by the current official dietary fiber methods provided appropriate corrections are made when needed (see below). New methods would have to be evaluated according to their ability to...

Fat Intake And Disease

There is a growing body of evidence that suggests high fat intake may specifically contribute to overeating and obesity, more so than other dietary energy sources like carbohydrates. Although fat is energy dense, it appears to have a weak effect on satiety (3). In contrast to the carbohydrates, fat oxidation seems to be is largely unrelated to fat intake, and dietary fats are more efficiently used and more readily converted to body fat (3). In fact, recent evidence suggests that carbohydrates may not make a significant contribution to body fat in humans under normal feeding conditions in humans (3). Finally, there is longitudinal data that point to a causal relationship between fat intake and weight gain over time in certain individuals (3). The Eskimos, for example, ingest mainly a high fat diet of fish, and experience less coronary heart disease and thrombosis than persons eating the high fat diets that contain mostly saturated fats (9). The major mechanism would seem to be the...

Development Of The Fat Replacer Market

In the 1980s, significant changes in the official nutritional recommendations were made when the relationship of carbohydrates and of fats to diet and health were better understood. Reports of the National Advisory Committee on Nutrition Education (NACNE) and the Committee on Medical Aspects of Food Policy (COMA) in the United Kingdom indicated that fat intake should be reduced from 42 , at the time, to 34 or 35 of total food energy in the diet (12,13). Further recommendations were to reduce the intake of saturated fat to 10 (NACNE) or 15 (COMA) of the food energy, to decrease salt intake, and to increase in the consumption of complex carbohydrates and dietary fiber (12,13).

Inulin And Oligofructose Dietary Fibers

In this chapter, I would summarize that both inulin and oligofructose are indigestible, have significant 'dietary fiber effects' often comparable to pectins, are carbohydrates of plant origin, naturally occurring in significant amounts and show beneficial effects on the gut flora. For these properties, no significant differences have been found between inulin and oligofructose.

Polydextrose Structure

Structure Polydextrose

The representative structure is derived from characterizations that include methylation and periodate oxidation for degree of branching Smith periodate degradation, methylation and acetolysis for linkage positions Smith degradation and acetolysis for anomeric linkage configuration and 13C-NMR (17). Polydextrose is more highly branched than other carbohydrates, such as the amylopectin component of starch, which contains mainly a-1,4 linkages with about 4-5 of a-1,6 linkages as branch points. The structural compactness and complexity of

Dietary Fiber Definition

Gums, algal and synthetic polysaccharides behave as DF. With the exception of lignin, dietary fibers are carbohydrates, and it has been suggested that the measurement of nonstarch polysaccharides is more accurate than determination of DF (22). On the other hand, it has been proposed that starch not digested in the small intestine and reaching the colon (i.e., resistant starch (RS)) is chemically and physically similar to other nondigestible polysaccharides and should therefore be included in the definition of DF (23). This definition describes polysaccharides and RS as a fiber by the AOAC assay (see later section).

Analysis Of Physiologically Functional Food Additives

Another class of complex carbohydrates which has been an important topic at this workshop are physiologically functional food additives such as inulin and oligosaccharide-based products. These materials are increasingly being used as fat substitutes, and for their beneficial health effects which are similar to those of dietary fiber, i.e. they are not digestible by human digestive juices, and cause an increase in the bifidobacteria population in the colon which in turn suppresses

The Food Label And Dietary Guidelines

The Dietary Guidelines for Americans which is issued every five years jointly by the Departments of Health and Human Services and of Agriculture represent fundamental, federal government public health policy. The fourth edition of the Dietary Guidelines recognizes the food label, especially the new Nutrition Facts label, along with the Food Guide Pyramid as the educational tools American consumers should use to put the Dietary Guidelines into practice. Consumers are advised to use the Nutrition Facts Label to choose a healthful diet (1, page 4). The same guidelines advise Americans to choose a diet with plenty of grain products, vegetables, and fruits . because they provide complex carbohydrates (1, page 22). However, for the nutrient complex carbohydrates the food label is not a very useful tool because complex carbohydrates do not appear in the Nutrition Facts panel. Eat foods with adequate starch and fiber. complex carbohydrate foods are better than simple carbohydrates Complex...

Postprandial Glucose Response

After a meal, the extent of rise of blood glucose depends on several interacting factors. At the beginning of this century, scientists assumed that the molecular size of the carbohydrate was an important determinant of its rate of digestion. Starch was thus considered to be digested and absorbed slowly whereas low-molecular-weight carbohydrates were believed to be absorbed rapidly, hence promoting high responses of plasma glucose. Studies have since shown that food properties other than molecular size influence the postprandial response and that starchy foods can have glycemic indices ranging from low to high (25-27). Any process that disrupts the physical or botanical structure of food ingredients will increase the plasma glucose and insulin responses (28). the metabolic response to starchy foods (cereals, legumes) and to foods containing low-molecular-weight carbohydrates (apple, orange, carrot) (31-40). Food structure is so important that the use of products with ''as fed'' food...


The issue of complex carbohydrates clearly has not been settled. Many in aca- 'S scientific issues associated with complex carbohydrates and, in light of these issues, address the rationale for adding complex carbohydrates to the nutrition label. (14) This workshop concluded that it is clear that this is not a simple task. Classifications cannot be made simply for chemical or analytical expediency if they do not express physiological significance. As a follow-up to the ILSI Conference the AOAC (Association of Official Analytical Chemists) International sponsored a Workshop in September 1995 with a goal of reaching a consensus on the definition of complex carbohydrates to support food labeling and dietary guidelines, to discuss state-of-the-art techniques in analytical methods, and to set

Caloric Value

Carbohydrates discussed concerns their caloric value. Indeed, being exclusive substrates for colonic fermentation, oligofructose and inulin have also much lower caloric value than equivalent but digested carbohydrates. Moreover, they are (at least) partly used as direct and indirect substrates for bacterial proliferation, a process that further reduces their bioavailability to the host. Based on experimental evidence as well as on basic biochemical knowledge a theoretical approach has been proposed to compare the metabolic efficiency (in term of ATP production) and thus the effective caloric value of a fructosyl moiety in chicory fructooligosaccharide and sucrose (as a typically digested substrate). As compared to sucrose chicory fructooligosaccharide has a relative caloric value of 25 to 35 , or 1 to 1.5 kcal g or 4 to 6 kJ g (16).


We feel that a more useful and practical solution at present is to use the terms starch and dietary fiber separately. An appropriate definition of starch follows Starch is a large polymer of glucose consisting of glucose units formed into unbranched amylose chains composed of alpha-1,4 linked glucose residues and highly branched amylopectin with alpha-1,4 and alpha-1,6 bonds. There are several methods to measure starch (4-9). There are commercially available kits to measure starch based on converting it to soluble fragments with minimal production of glucose followed by quantitative hydrolysis by amyloglucosidase (4). (This kit can also provide an estimation of enzyme- resistant starch). In the method for total starch, starch is solubilized with DMSO or the sample is cooked in buffer with a thermostable alpha-amylase. Complete breakdown to dextrins is carried out by subsequent treatment with the de-branching enzyme pullulanase in combination with alpha-amylase to minimize the amount...

Dietary Fiber

To consider dietary fiber as complex carbohydrates also ignores the fact that the non-carbohydrate components participate in maintaining the botanical (plant cell wall) structure of dietary fiber and the structure of the food. The botanical structure and particle size may account for a significant proportion of the beneficial effects of dietary fiber on postprandial blood glucose response and intestinal function (11).

Panel Discussion

What are the important scientific reasons for including complex carbohydrates on the label The panel emphasized the need to increase the physiologic database supporting the overall benefits of a diet high in complex carbohydrates to support and substantiate the current dietary recommendations and reach a consensus on the definition and methodology to reduce the research fragmentation that currently exists. The panel suggested practical reasons for putting the term complex carbohydrate on the label 1. Complex carbohydrates are a marker of foods people should be consuming. S 2. Use of the term is a normal progression of knowledge about the health benefits of carbohydrates complex carbohydrates and even types of complex carbohydrates (such as resistant starch) are an important component of the diet. It was noted that this is similar to the progression of knowledge on the importance of fiber and on specific types of fiber (for example, soluble and insoluble fiber). 2. Given the current...

Preface And Overview

The objectives for the workshop were 1) To evaluate complex carbohydrates from the perspectives of definition and methodology, nutrition and health, and dietary guidelines and public health 2) To evaluate critically the scientific issues associated with the possible addition of complex carbohydrates to the nutrition label and 3) To explore issues associated with developing a rational public policy for the labeling of complex carbohydrates. Since the 1970s, complex carbohydrates have been an important component of various dietary guidelines. However, the term has been used without much attention given to definition, analytic methodology, or physiologic effects. These issues must be resolved before complex carbohydrates can meet the criteria for use in food labeling. 2. Can analytic methodology be developed that reflects all of the components encountered in the broad definition of complex carbohydrates 3. Is there a clear understanding of the physiologic benefits of complex...


An analytical method was developed to measure complex carbohydrates (CC) in foods as the sum of available starch and total dietary fiber. Desugared food samples were subjected to sequential starch and protein digestion by using heat stable alpha-amylase, protease, and amyloglucosidase, and filtered. Filtrate containing enzymatic digestate of starch was analyzed for glucose and maltose by using a high pressure liquid chromatography (HPLC) technique. Residues were subjected to further analysis of dietary fiber by using a gravimetric method. The available starch values obtained by the HPLC method were precise and in agreement with those derived by the calculation method ( Available Starch Total Carbohydrate - Total Dietary Fiber - Sugars - H2O - Protein - Fat - Ash). The complex carbohydrates values generated as the sum of analytically measured available starch and total dietary fiber were precise and agreed with those calculated by the difference method ( CC Total Carbohydrates - Sugars...


Complex carbohydrates are determined as the sum of available starch and dietary fiber. Duplicate samples of dried foods, fat-extracted if containing 10 fat, undergo sequential enzymatic treatments by heat-stable a-amylase, protease, and amyloglucosidase to digest starch and protein. Enzymatic digestion steps have been adopted from AOAC official final action methods 991.43 (17, 18) and 985.29 (18, 19). For available starch, the enzyme digestate, containing mostly monosaccharides and some disaccharides, is analyzed by HPLC. Measured glucose and maltose are converted into a starch unit and reported as available starch. HPLC conditions have been adopted from AOAC official final action method 982.14 (18) for sugar determination. For total dietary fiber (TDF), the enzyme digestate is treated with alcohol to precipitate soluble dietary fiber before filtering, and TDF residue is washed with alcohol and acetone, dried, and weighed. TDF residue values are corrected for protein, ash, and blank....

Analytical Results

The HPLC method described in this paper is a quantitative method for available starch in foods. A variety of samples, such as grains, ready-to-eat breakfast cereals, breads, crackers, fruits, vegetables, and mixed diets can be analyzed by using this method. Complex carbohydrates are determined as the sum of available starch and dietary fiber available starch by high performance liquid chromatogra-phy determination of glucose and maltose present in the enzymatic hydrolysates of desugared sample and dietary fiber by enzymatic gravimetric method, official final action method 991.43. Complex Carbohydrates 141 Table 7 Determination of Available Starch in Foods (by HPLC) and Complex Carbohydrates*1*2 Table 7 Determination of Available Starch in Foods (by HPLC) and Complex Carbohydrates*1*2


Structural protein (46-47) and lignin are non-carbohydrate components of dietary fiber. One method (Englyst) overlooks lignin and only one method (Mongeau) measures the small amount of undigestible protein. Permanganate lignin and Klason lignin values are not always in agreement (43-44). Klason lignin is generally higher and more variable than permanganate lignin. Lignin is difficult to separate from some hemicelluloses and can react with protein during food processing (48). The rapid Mongeau method includes treatments that represent an appropriate preparation for measuring permanganate lignin in foods, and values are little influenced by food processing. According to Selvendran (7), the acetyl bromide method needs only a 5-10 mg sample but comparison of values has not been made. Lignin is regarded as a minor component of most plant foods but the total polyphenolic material (including lignin) represents 5-20 of the dry cell wall (7).

Chemical Treatments

Peroxide bleaching frees carbohydrates bound to lignin. Bleaching increased soluble fiber neutral sugars in oat hulls by over 3 , while lignin decreased over 6 compared to non-bleached hulls (18). Health benefits due to phenolic compounds and other phytochemicals associated with fiber in foods may be lost during bleaching (19). As more consumers appreciate the importance of fiber in their diets, there may be less need to bleach fiber-rich materials to enhance marketability. involvement of these carbohydrates in the formation of Maillard polymers is most likely responsible for the higher apparent lignin formation.

Rice Bran

The unsaponifiable fraction consists of compounds such as oryzanols, beta-sitosterol, and tocopherols, which may have cholesterol-lowering activity. The major carbohydrates in commercial rice bran are cellulose, hemicellulose, and starch. Starch is not present in the outer pericarp layers, but because of endosperm breakage during milling it appears in the bran. The quantity varies according to the amount of breakage and degree of milling, but values of 10-20 percent are typical. Hemicellulose and cellulose have been reported to comprise 8.7-11.4 percent and 9.6-12.8 percent of the bran. Beta-glucans in the bran are present at levels of less than 1 percent.

Characterization Of The Protein Product

Tate dehydrogenase), or as a therapeutic agent (e.g., insulin and growth hormone). However, it is normal, when a protein has been isolated for the first time, to characterize it in terms of structure and function. Several features are generally expected in characterization of a new protein. These include molecular weight, or at least the size of the subunit(s), determined by SDS-polyacrylamide gel electrophoresis (Chapter 10) and or gel filtration (unit 8.3). Spectral properties such as the UV spectrum (Trp and Tyr content), circular dichroism (CD) spectrum (secondary structure), and special characteristics of proteins with prosthetic groups (e.g., quantitation and spectra) may be presented. The quantity and nature of carbohydrates on glycoproteins should be determined (Chapter 12). Also, if the gene has not already been reported, some amino-terminal sequence analysis should be given, if at all possible, along with the results of a database search for similar sequences (unit 2.1)....

Scope Of The Book

The overall scope of this book is the implementation and application of available theoretical and computational methods toward understanding the structure, dynamics, and function of biological molecules, namely proteins, nucleic acids, carbohydrates, and membranes. The large number of computational tools already available in computational chemistry preclude covering all topics, as Schleyer et al. are doing in The Encyclopedia of Computational Chemistry 23 . Instead, we have attempted to create a book that covers currently available theoretical methods applicable to biomolecular research along with the appropriate computational applications. We have designed it to focus on the area of biomolecu-lar computations with emphasis on the special requirements associated with the treatment of macromolecules.

Production of Monoclonal Antibodies

Highly specific antibodies can be obtained by fusing immune B cells from the spleen with tumor cells to produce hybridomas, each of which will then secrete a single antibody. The desired antibody-producing hybridoma can be identified by a screening process. If this hybridoma is subjected to a cloning process in which clones are selected, such that all progeny are derived from a single cloned parental cell, a monoclonal antibody is obtained. Monoclonal antibodies have high specificity and can be produced in large quantities. Thus, these biological reagents have been used extensively as probes in a wide range of systems including the characterization of novel cell-surface and soluble proteins and carbohydrates, as enzyme catalysts, and for targeting in immunotherapy (see commentary).

Primary Nursing Diagnosis

Because of the negative effect of physical and emotional stress on the patient with adrenal insufficiency, promote strategies that reduce stress. Teach the patient to rest between activities to conserve energy and to wear warm clothing to increase comfort and limit heat loss. To limit the risk of infection, encourage the patient to use good hand-washing techniques and to limit exposure to people with infections. To prevent complications, teach the patient to avoid using lotions that contain alcohol to prevent skin dryness and breakdown and to eat a nutritious diet that has adequate proteins, fats, and carbohydrates to maintain sodium and potassium balance.

Overview of Available Force Fields

Treat small molecules in the gas phase, although exceptions do exist. Such force fields may also be used for biological simulations however, the lack of emphasis on properly treating biological systems generally makes them inferior to those discussed in the previous paragraphs. The optimized potential for liquid simulations (OPLS) force field was initially developed for liquid and hydration simulations on a variety of organic compounds 28,29 . This force field has been extended to proteins 30 , nucleic acid bases 31 , and carbohydrates 32 , although its widespread use has not occurred. Some of the most widely used force fields for organic molecules are MM3 and its predecessors 33 . An MM3 force field for proteins has been reported 34 however, it too has not been widely applied to date.

The Basics of Nutrition45

They are all made up primarily of three components carbohydrates, protein, and fats. All three of these components contain calories, meaning that they produce energy in your body. In addition to carbohydrates, protein, and fats, your body needs other nutrients, including vitamins, minerals, and water. Together these categories of nutrients are known as the building blocks of nutrition. Carbohydrates Carbohydrates supply the main source of energy for your body, so many nutritionists recommend that they should make up the majority 50 or 60 percent of your intake of calories. Carbohydrates consist of the starches, sugars, and fiber found in foods that come from plants. There are two types of carbohydrates simple and complex. Simple carbohydrates, also known as simple sugars, taste sweet and are quickly absorbed and digested. Examples of simple carbohydrates are table sugar, honey, corn syrup, and the type of sugar found in fruit. Complex carbohydrates refer to the starches or fiber found...

Factors affecting body acidbase balance

* Body acid production creates a hydrogen ion load, mainly from oxidation of proteins, carbohydrates and other organic molecules. This amounts to 50-100 mmol of hydrogen ions per day in the adult, and includes volatile acids (such as carbonic acid) and fixed acids (such as phosphoric and sulphuric acids). The body is a net producer of acid. Acids produced by the body consist of carbonic acid (respiratory acid), and various metabolic acids, including sulphuric acid, phosphoric acid, lactic acid, citric acid, ammonium ions and ketone bodies (acetoacetic acid and beta-hydroxybutyric acid).

Lipid Droplets Hyaline Cartilage

Oo Paraplasmic substances (cell inclusions, stored materials) are either derived ID from metabolic activities (storage materials) or from the enclosure of dead substances (metabolic residue or phagocytosed materials ). Carbohydrates, proteins, fat (mostly triglycerides), ferritin (iron storage particles) and pigments are among the most important paraplasmic inclusions. This figure shows four isogenous cartilage cells (two chondrons or territories) with fat droplets of different sizes. These fat droplets (stained red) are characteristic of mature chondrocytes (see Fig.61). The cell nuclei are stained blue. There is a narrow border between the cartilage cells, which stands out because of its high refractive index. This is the cartilage cell capsule , which is part of the territorial extracellular matrix (see Figs. 193-198).

Monitoring of the Fetus in a Mother with Graves Disease

(a) Hyperemesis gravidarum is common and around 5 of cases require hospital admission because of dehydration and ketosis. Thyroid function should be checked in these patients a correlation has been established between the severity of the hyperemesis and thyroid function with an elevated FT4 and FT3

Posttranslational Modifications And Expression System Choice

The most common issue is the need to obtain glycosylated proteins. The majority of proteins secreted by mammalian and yeast cells are glycoproteins. Carbohydrates are attached to proteins at asparagine residues in the case of N-linked glycosylation, or to serine threonine residues in the case of O-linked glycosylation (see Chapter 12 for further information).

Immunocytochemical Characterization of hES Cells

The panel of surface markers used to characterize undifferentiated hES cells recognize specific proteins or carbohydrates expressed on the cell surface. Common markers of undifferentiated hES cells include the globoseries glycolipid antigens designated stage-specific antigen-1 (SSEA-1), SSEA-3, and SSEA-4 Kannagi et al., 1983 and the keratin sulfate-related antigens (Trafalgar antigens) TRA-1-60 and TRA-1-81 Andrews et al., 1984 . Other markers such as the POU transcription factor OCT-4 are also expressed in pluripotent cell populations including undifferentiated hES cells Rosner et al., 1990 . Furthermore, it has been shown that OCT-4 expression is necessary to maintain pluripo-tency in ES cells Nichols et al., 1998 Boyer et al., 2005 , and therefore the loss of OCT-4 expression can be used as a marker of differentiation Loh et al., 2006 .

Influence of Suspending Medium Composition on Survival of Live Cells to Freeze Drying

Attempts to freeze-dry cells in water or a simple salt solution typically result in poor survival. A wide range of protective media has been developed for preserving freeze-dried vaccines, including augmented growth media or sugar solutions. Carbohydrates are widely used as freeze-drying protectants either individually or in combination with other solutes. They should be chosen on the basis of experimentally determining their freeze-drying characteristics rather than on a pragmatic basis. Monosaccharides, such as glucose, provide good bioprotection during freezing and freeze-drying but exhibit low glass transition (Tg') or collapse temperatures (Tc) and dry with collapse when orthodox freeze-drying cycles are used. Disaccharides are effective freeze-drying protectants, and because they display higher collapse than monosaccharides, freeze-dry successfully when conventional drying cycles are used. Reducing sugars may induce damaging Maillard reactions, thereby compromising stability,...

Bacterial Expression Of Proteins Normally Glycosylated

Intermediates, and thus enhance folding, while not necessarily affecting the stability of the native state (Kern et al., 1993, and references cited therein). In eukaryotic cells, interference with protein glycosylation can lead to the formation of misfolded, aggregated, and degraded protein. This indicates that in vivo glycosylation (N-linked) may also prevent the aggregation of folding intermediates (reviewed by Helenius, 1994). Detailed NMR studies on glycoproteins have clearly shown that carbohydrates stabilize folded proteins and even prevent marginally stable proteins from unfolding (for a review, see Wyss and Wagner, 1996).

Purification and Characterization of Recombinant Produced Human APC

Unlike the other posttranslational modifications described above, the carbohydrates on rHPC differed from those on plasma-derived HPC. The structures of oligosaccharides on recombinant glycoproteins are determined to a large extent by the cell line used 32 . Oligosaccharides found on glycoproteins can influence parameters that are important to drug design, such as circulatory clearance, tissue targeting, immunogenicity, and efficacy 32 . In view of this, a detailed analysis of the oligosaccharide structures on rHPC from HEK293 cells was performed 9,33 , and novel and rare Asn-linked oligosaccharides were identified (Figure 4.4). The novel trisaccharide epitope on rHPC was designated the PC293 determinant, and, along with the two rare disaccharide epitopes, GalNAcP(1 4)GlcNAcP(1 4) and NeuAca(2 6)GalNAcP(1 4), have been found in naturally occurring human glycoproteins 34-37 . Glycosyl composition analysis has shown that these GalNAc-containing rare oligosaccharides are not present in...

Activated metabolites

ATP and the other nucleoside triphosphate coenzymes not only transfer phosphate residues, but also provide the nucleotide components for this type of activation reaction. On this page, we discuss metabolites or groups that are activated in the metabolism by bonding with nucleosides or nucleotides. Intermediates of this type are mainly found in the metabolism of complex carbohydrates and lipids.

Protein Fractionation

The GF fractionation buffer must be selected in light of the method planned for testing purity of the final proteinfraction (step 11), as this may put special restrictions on the buffer (e.g., sodium azide absorbs light at 254 nm and will also interfere with the anthrone reaction used for assaying carbohydrates).

Composition of Base Matrix

The support (base matrix) upon which the ligand is immobilized contributes to the selectivity of the HIC medium. The two most common base matrices in use for HIC are 4 and 6 cross-linked agarose, which are strongly hydrophilic carbohydrates, and the synthetic copolymer materials. The selectivity of an agarose-based matrix and a polymer-based matrix substituted with the same ligand will be different. It is possible to achieve the same results using either type of support, but only if the the adsorption and elution conditions are modified appropriately.

Packing A Column With An Hic Matrix Of 90m BEADS

This protocol describes preparation and degassing of the HIC gel medium and packing of the column. After these procedures are completed, a packed column will be ready for testing (Basic Protocol 2) and sample application. This protocol and that for high-resolution media (see Alternate Protocol) are generally applicable to all bulk carbohydrate-based media, and can be skipped if a prepacked column is to be used. If non-carbohydrate-based media are to be used, consult the manufacturer's literature for packing instructions.

The Properties Of Peptides And Proteins And Their Implications For Hplc Method Development

The 20 naturally occurring L-a-amino acids found in peptides and proteins vary dramatically in terms of the properties of the side chain or R-groups. Table 8.7.1 lists some of the fundamental properties of the common L-a-amino acids found in peptides and proteins. This chemical diversity becomes even greater in circumstances where some of these side chains have been post-translationally modified with carbohydrates or lipid moieties. The side-chains are generally classified according to their polarity (e.g., non-polar or hydrophobic versus polar or hydrophilic). The polar side chains are divided into three groups uncharged polar, positively charged or basic, and negatively charged or acidic side chains. Peptides and proteins generally contain several ionizable basic and acidic functionalities. They therefore typically exhibit characteristic isoelectric points with the overall net charge and polarity in aqueous solution varying with pH, solvent composition and temperature. Cyclic...

Respiration and fermentation

In the presence of oxygen (i.e., in aerobic conditions), most animal cells are capable of respiring various types of nutrient (lipids, amino acids, and carbohydrates) i. e., using oxidative processes to break them down completely. If oxygen is lacking (i.e., in anaerobic conditions), only glucose can be used for ATP synthesis. Although in these conditions glucose breakdown in animals already ends in lactate and only produces small quantities of ATP, it is decisively important for the survival of cells at times of oxygen deficiency.

Cell Adhesion And Labeled Pathways

Neural cell adhesion molecule (NCAM), the earliest identified of the CAMs, appears to be expressed on all vertebrate neurons and glia (Edelman, 1984). NCAM exists in many different forms, some with an intracel-lular domain that may interact with the cytoskeleton and some without. The extracellular portion of NCAM can be highly modified by the addition of carbohydrates, particularly sialic acid residues. Nonsialylated forms are very adhesive compared to the sialylated forms. In the course of a neuron's development, there may be changes in the sialylation state of its NCAM, thus adjusting its adhesion (Walsh and Doherty, 1997). For example, developing motor neurons of the chick grow out of the spinal cord and enter a complicated plexus region, where they cross in many directions and eventually segregate into distinct nerve roots leading to their appropriate muscles. During the time when these axons are growing in the plexus, the NCAM they express is highly sialylated. This keeps the...

Capillary Electrophoresis of Proteins and Peptides

Capillary electrophoresis (CE) is a high-resolution technique for the separation of a wide variety of molecules of biological interest such as metabolites, drugs, amino acids, nucleic acids, and carbohydrates. This unit focuses on the use of CE to separate proteins and peptides. As with polyacrylamide gel electrophoresis (PAGE unit 10.1), CE separations of proteins and peptides are based on charge-to-mass ratios. While PAGE separations are restricted to polyacrylamide matrices and a relatively small number of buffer systems CE separations can be achieved in a variety of different matrices using a wide range of electrophoresis buffers. Consequently, there is a much greater flexibility in the design of optimal separation protocols.

Discharge And Home Healthcare Guidelines

Magnesium plays an important role in neuromuscular function. It also has a role in several enzyme systems, particularly the metabolism of carbohydrates and proteins, as well as maintenance of normal ionic balance (it triggers the sodium-potassium pump), osmotic pressure, myocardial functioning, and bone metabolism. Because the kidneys are able to excrete large amounts of magnesium ( 5000 mg day), either the patient has to ingest extraordinary amounts of magnesium or the glomerular filtration of the kidneys needs to be very depressed for the patient to develop hypermagnesemia. Complications include complete heart block, cardiac arrest, and respiratory paralysis.

Pharmacologic Highlights

Never force an unconscious or semiconscious patient to drink liquids because of the risk of aspiration into the lungs. Continue to repeat the oral intake of carbohydrates until the blood glucose rises above 100 mg dL, and administer the next meal as soon as possible. If the next scheduled meal is not ready for more than 30 minutes or longer, provide the patient with a combination of carbohydrate and protein, such as V2 cup milk, 1 ounce of cheese, and three saltine crackers.

Determining the Structure of Glycan Moieties by Mass Spectrometry

Mass spectrometry provides a complementary method to HPLC (unit 12.6) for the analysis of glycans. In general, its resolution, defined as its ability to detect carbohydrates in a complex mixture, is higher than in HPLC, particularly for larger structures. In addition, it provides a mass from which the composition of the glycan, in terms of its isobaric monosaccharide composition, may usually be determined directly. Analysis by mass spectrometry is also faster than by HPLC. On the other hand, mass spectrometry does not usually allow isomeric compounds to be distinguished unless they give diagnostic fragmentation spectra, and, because mass spectrometry is more susceptible to the presence of salts and residual buffers, efficient sample preparation is much more critical and sensitivity generally lower. The high sensitivity and specificity of HPLC is a result of the fluorescent derivatives that are necessary for detection. Although the same technique of derivative preparation can be...

Analysis Of Glycans By Electrospray Ionization Mass Spectrometry

The glycan solution should be as free from other compounds as possible, as these compounds may compete for the ionization and reduce the signal strength. Electrospray is not as sensitive for detection of carbohydrates as it is for peptides, and concentrations of 1 to 10 pmol l solvent are typically needed. However, certain reducing-terminal derivatives, such as that formed from the N,N-diethylaminoethyl ester of 4-aminobenzoic acid (ABDEAE derivatives Mo et al., 1999) provide increases in sensitivity on the order of 100- to 1000-fold over that of the underivatized glycan. Such derivatives may be prepared in the same way as the 2-aminobenzamide derivatives with the appropriate amine substituted for 2-AB. Glycans derivatized with 2-AB or 2-AMAC (Charlwood et al., 1999) may be cleaned (see Support Protocol 3) and then dissolved in 1 1 (v v) methanol water or 1 1 (v v) acetonitrile water containing 1 to 50 mM sodium iodide. Sodium iodide is used to produce M + metal + ions 0.1 formic acid...

DRG Category 296 Mean LOS 54 days Description Medical Nutritional and

Magnesium is regulated by vitamin D-regulated gastrointestinal (GI) absorption and renal excretion. Magnesium plays an important role in neuromuscular function. It also has a role in several enzyme systems, particularly the metabolism of carbohydrates and proteins, as well as maintenance of normal ionic balance (it triggers the sodium-potassium pump), osmotic pressure, myocardial functioning, and bone metabolism. Deficits of magnesium lead to deficits in calcium, and the two electrolyte imbalances are difficult to differentiate. The hypocalcemia that accompanies hypomagnesemia cannot be corrected unless the magnesium is replaced. Hypo-magnesemia is also a stimulus for renin release, which leads to aldosterone production, potassium wasting, and hypokalemia. Because magnesium regulates calcium entry into cells, consequences of magnesium deficiency include ventricular dysrhythmias, an enhanced digitalis toxicity, and sudden cardiac death. Deficits in potassium and calcium potentiate the...

Control For Olinked Glycosylation

Traditionally, mild alkaline reduction (reductive p-elimination) has been used to release O-linked carbohydrates from proteins (Amano and Kobata, 1990). This method has been adapted for blots to show that lectin antibody reactivity is toward O-linked rather than N-linked glycans (Duk et al., 1997). Proteins blotted to PVDF are treated with 55 mM NaOH overnight (releasing O-linked sugars) and then probed using lectins or antibodies.

Characterization Of Labeled Glycans By Pelimination And Chromatography

This protocol has three steps (1) the release of carbohydrates as sugar alditols by reductive P-elimination (2) desalting the sample, while confirming the size of the labeled sugar alditol(s) and (3) confirmation that the product is 3H Gal1-4 GlcNAcol (from galac-tosyltransferase labeling) or 3H GlcNAcol (from metabolic labeling).

Management of the Metabolic Syndrome

Healthy lifestyle promotion includes (1) dietary changes in terms of moderating calorie intake to achieve a 5-10 loss of body weight in the first year and changing dietary composition (2) increasing physical activity and (3) changes in dietary composition to lower intake of saturated fats, trans fats and cholesterol, and include carbohydrates with a low glycemic index and high content of soluble fiber.

Critical Parameters and Troubleshooting

Product analysis is critical for metabolic labeling with glucosamine. While UDP-GlcNAc is the major product, glucosamine can enter other biosynthetic pathways, such as those used for amino acid synthesis. This issue was highlighted by studies of the SV40 large T-antigen. Some researchers have found that the SV40 large T-antigen labels with a number of different tritiated carbohydrates. However, O-GlcNAc is the only carbohydrate post-trans-lational modification of the SV40 large T-anti-gen. The incorporation of glucosamine into amino acid biosynthetic pathways could be reduced by growing cells in the presence of excess nonessential amino acids (Medina et al., 1998).

Development of Ceredase

Since P-glucocerebrosidase is a large glycoprotein and carbohydrates are essential for correct folding and activity, a mammalian expression system was chosen. The cDNA for P-glucocerebrosidase was cloned into CHO cells using a dihydrofolate reductase (DHFR) expression system. Following amplification, highest producing clones were adapted to an anchorage-dependent serum-free microcarrier spinner culture system for production evaluation 61 . The highest producing line was then progressively scaled up from spinner flasks to production bioreactors of increasing size up to 2000 L. The bioreactors were operated in a continuous perfusion mode to maximize production efficiency and reduce the potential for proteolytic degradation or other posttranslational modifications of the product due to cell lysis. The Cerezyme purification process was similar to that used for the production of Ceredase and was based on standard chromatographic procedures. Since the biore-actors were harvested in a...

Infancy And Childhood

This is maintained further by weaning on to a high-fat low-carbohydrate diet but falls rapidly with weaning on to the normal high-carbohydrate diet.5'18 However, during this transition there are significant nutritional and metabolic differences between human and rat. In rat plasma fatty acid substrates for ketogenesis come mainly from hydrolysis of maternal milk triacylglycerols, due to lack of white adipose tissue at birth. In human neonates there is significant accumulated triacylglycerol in the liver, rapidly mobilized for utilization in situ after birth. Significant amounts of fat are stored in the human fetus in white adipose tissue comprising 16-20 body mass, mainly as triacylglycerides containing high proportions of palmic (C16) and oleic (C18 1) acids. Plasma free fatty acids start to rise soon after birth from lipolysis of this store. Once lactation is established, fatty acids from intestinal hydrolysis of milk triacylglycerols are directly...

Mechanisms of action

Hypoglycemia in acarbose- or miglitol-treated subjects must be treated with simple carbohydrates found in milk, juices, or glucose tablets. Disaccharides or polysaccharides (sucrose (table sugar), candy, and soft drinks) cannot be used because the a-glucosidase inhibitory effects delay their hydrolysis and absorption. When used as monotherapy, acarbose and miglitol are not associated with hypoglycemia or significant weight changes. Blocking the absorption of complex carbohydrates decreases the caloric uptake of the small intestine, but the large intestine compensates to assure that adequate caloric goals are met. a-Glucosidase inhibitors do not significantly affect LDL or HDL cholesterol concentrations, but triglyceride levels decline. These agents may prove to be useful in the management of severe hypertriglyceridemia in both the diabetic and non-diabetic population.

Key Ionization Methods And Related Ancillary Techniques

The decade of the 1980s marked a revolutionary period in the development of soft ionization techniques for the analysis by mass spectrometry of large, polar, nonvolatile molecules (Carr et al., 1991). These ionization techniques produced primarily intact protonated molecular ions of peptides and proteins. In the early 1980s, fast atom bombardment (FAB) and, to a lesser extent, plasma desorption (PD) mass spectrometry were used extensively for structural characterization of peptides and small proteins. In 1988, electrospray ionization (ESI) and matrix-assisted laser desorp-tion ionization (MALDI) mass spectrometry were first shown to be useful for the analysis of peptides, proteins, carbohydrates, and oligonucleotides. Within a few years of their introduction, ESI and MALDI techniques became the MS methods of choice for biopolymers analysis, largely supplanting FAB-MS and PD-MS. For this reason these techniques will be the focus of this overview. The capabilities and sample...

Ketone Body Concentrations within Brain

Since the liver can achieve a plasma ketone body concentration of between 1-2mM24 when exposed to a high fat diet, what might be the advantage of an organ such as brain in possessing an endogenous ketogenic capacity Firstly, it might provide a compensatory mechanism when, for reasons of reduced hepatic ketogenesis, plasma ketone body concentrations fall to lower limits (eg. 0.5 mM). This would hold true if there was an absolute requirement for a given concentration of ketone body within brain eg. 1 mM. However, there is some evidence to suggest that the brain as a whole uses ketone bodies facultatively, rather than having an obligate requirement for a minimum plasma concen-tration.25 Secondly, however, possessing an endogenous ketogenic capacity might expose ketolytic cell-types in the brain to ketone body concentrations that are simply unattainable in the plasma as a result of hepatic ketogenesis. This is potentially very important in the brain, since ketone body concentrations in...

Ketone Bodies and Seizure Disorders

Epileptic seizures affect approximately 1 of children and constitute one of the most common paediatric neurological disorders. An increasingly popular epilepsy treatment is the so-called ketogenic diet (KD),28,29 a high fat diet with associated high plasma concentrations of free fatty acids, ketone bodies and glucocorticoids,24 which can result in prolonged seizure remission in up to 30 of intractable infant epilepsies.28,29 Although the biochemical basis behind many seizure-types remains to be defined, certain anti-epileptic drugs (AEDs) (eg. vigabatrin) appear to enhance the activity of the seizure-suppressing inhibitory neurotransmitter yaminobutyric acid (GABA), whilst others (eg. lamotrigine, felbamate) appear to inhibit the activity of seizure-inducing excitatory amino-acids (EAAs) such as aspartate and glutamate.30 Similarly, the mode of action of the KD appears to involve alterations in amino-acid neurotransmitter concentrations, since, as previously mentioned, addition of...

Assays for the Mannan Binding Lectin Pathway

As discussed in previous units of this chapter, the complement system is part of the innate immune system and contributes to the establishment of adaptive immune responses. The mannan-binding lectin (MBL) pathway of the complement system is activated when MBL binds to specific carbohydrates (e.g. mannose, glucose, L-fucose, and N-acetylglu-cosamine) expressed on the surfaces of microorganisms. Such sugar residues are usually present at the terminal nonreducing position. High-affinity binding with MBL requires that these sugars be clustered to activate the MBL pathway. The affinity towards a single monosaccharide is in the order of 10-3 M, whereas the avidity can reach 10-9 M when multiple bindings are achieved. In the circulation, MBL is found in a complex with serine protease zymogens MASP-1 (MBL-associated serine protease-1), MASP-2 (MBL-as-sociated serine protease-2), and MASP-3 (MBL-associated serine protease-3). The MBL-MASP complex also contains a nonenzymatic protein MAp19. The...

Characteristics of Campylobacter and Arcobacter species

Campylobacters are Gram-negative, micro-aerophilic, non-sporing, small vibroid (spiral-shaped) cells that have rapid, darting reciprocating motility. They reduce nitrate and nitrite (apart from C. jejuni subsp. doylei and C. fennelliae) and are unable to oxidise or ferment carbohydrates. Campylobacter jejuni is the most significant of the four thermophilic Campylobacter species and is one of 20 species and subspecies within the genus Campylobacter and family Campylobacteraceae, which also includes four species in the genus Arcobacter. The other thermophilic species include C. coli, C. upsaliensis and C. lari. The thermophilic species are characterised by their ability to grow best between 37 and 42 C and their inability to grow at 25 C. Strains of the two genera Campylobacter and Arcobacter have similar morphology and metabolism, and share several other genotypic and phenotypic features.

Determining the CD Spectrum of a Protein

Many of the complex phenomena associated with food processing, including cooking, involve the interactions of biopolymers. These interactions are frequently triggered by changes in the three-dimensional conformation of the biopolymers, which may be caused by changes in temperature, pH, or ionic strength, or by modifications made by hydrolytic enzymes. The understanding of such processes, necessary for their control, requires knowledge of the dependence of the conformational changes and interactions on environmental conditions. Circular dichroism (CD) can provide a sensitive indicator of these changes at the molecular level (Li-Chan, 1998), provided the sample is in the form of an optically clear solution and, for most applications, contains purified components. This unit reflects the fact that CD has been used mainly to study proteins, but it can, in principle, be used for any molecules containing a chromophore that absorbs radiation in an accessible region of the spectrum, such as...

Characteristics of the genus Yersinia 1461 Introduction

The genus Yersinia comprises Gram-negative, rod-shaped bacteria, which are capable of both oxidative and fermentative metabolism of carbohydrates. The genus is a member of the family Enterobacteriaceae, although it has a number of distinctive characteristics, including small colony size and, in many circumstances, coccoid morphology of cells. Yersinias are also considered as 'low-temperature' pathogens and, unlike most other members of the Enterobacteri-aceae, are able to grow at 4 C. Incubation temperature has a profound effect on growth characteristics between 28-30 C and 37 C (Sheridan et al., 1998). Biochemical characteristics and suppression of expression of cellular components are involved. The genus comprises environmental organisms, which may be oppor

Background Information

The BB rat strain was derived from a colony of outbred Wistar rats that developed spontaneous diabetes mellitus at the BioBreeding Laboratories, Ottawa, Canada, in the 1970s. Affected animals became the founders of the inbred diabetes-prone (DP)-BB Wor rat strain used in the majority of published studies. At the sixth generation of inbreeding, a subpopulation of nondiabetic DP-BB Wor rats was selected to start a control line. Now designated as diabetes-resistant (DR)-BB Wor rats, these coiso-genic descendants of DP-BB Wor forbearers do not develop spontaneous diabetes. Both BB Wor rat lines are now fully inbred. BB Wor and all other BB rat strains express the RT1u rat major histocompatibility complex (MHC) haplotype. As is the case with human insulin-dependent diabetes mellitus (IDDM), predisposition to the disease is strongly linked genetically to MHC genes. The onset of hypergly-cemia is preceded by mononuclear cell infiltration of the islets (insulitis) and the appearance of...

Quality Control Evaluations

Extensive analysis of rFIX is undertaken in accordance with industry standards for recombinant proteins to assess the (1) identity, (2) purity, (3) potency, and (4) safety and quality of the rFIX product obtained from the cell culture and purification processes (the drug substance) and also of the final formulated enzyme product (Section 11.5.5) 59 . In terms of purity, rFIX consists of both zymogen (mature, inactive FIX) and activated products of the mature zymogen. The cleavage products of both pdFIX and rFIX preparations are similar, with both containing FIX and FIX-related species (such as FIXa), although the amounts in each type of preparation may vary. Any non-FIX species derived from host cells or purification processes (including host cell proteins, carbohydrates, and small molecules) are considered impurities 59 .

RFIX Biochemical Properties and Characterization

Within the EGF-like domains, the carbohydrates structure and content, as determined by peptide mapping and mass spectrometry (MS) (O-linked glycosylation at S53 and S61) are the same in rFIX and pdFIX, with some minor differences. The relative proportion of P-hydroxylation of D64 was slightly less in rFIX than

Identifying Adulterants and Diluents

The simplest way to identify diluents in controlled substance mixtures is by microscopic identification. Common diluents along with the sugars carbohydrates starches described above include sodium chloride, calcium carbonate, and various types of amorphous materials. Because of their optical properties, these materials lend themselves well to a microscopic identification. Chemical separations are fairly easy because these materials are usually insoluble in solvents such as diethyl ether or hexane, and slightly soluble in solvents such as methanol. Most organic materials are soluble in methanol or some other polar solvent. The sugars carbohydrates starches can be further identified using IR following the separation if only one sugar is present. If not, HPLC can be used to identify the sugars.

Glucocorticoids and Fat Mobilisation a Response to Stress

We can more easily understand the logic of the synergistic interactions of the GR and PPAR RXR signal pathways on mHS gene expression, when we realise that one of the functions of glucocorticoid release into the blood is in the response of starvation stress.21 Starvation partly parallels the situations of the suckling neonate or the adult on a high fat low carbohydrate diet in that all three situations result in elevated plasma free fatty acid concentrations.22,23 In the case of starvation, plasma free fatty acid concentrations increase as a result of fat mobilization from adipose tissue, in an attempt by the body to compensate for low blood glucose fuel availability. Such fatty acids are predictably converted to ketone bodies, via the hepatic HMG-CoA cycle, resulting in a significant hyperketonaemia that can provide up to 70 of the fuel requirements of the brain during starvation.22,23 Glucocorticoids released from the adrenal cortex act to effect such increases in plasma free fatty...

The ABO and Rhesus Systems

Abo Incompatibility Newborn

The discovery of the ABO system at the end of the nineteenth century laid the foundation for clinical transfusion practice. It is now known that ABO antigens expressed on red cells are determined by genes located on the long arm of chromosome nine. These genes code for glycosyl transferases, which attach different carbohydrates (sugars), to a terminal galactose of an oligosaccharide chain. These oligosaccharide chains are attached to phospholipids in the red cell membrane and to proteins (glycoproteins) in plasma. The ABO system is illustrated in Figure 6.1. In blood group A, the terminal sugar is N-acetyl D-galactosamine and in blood group B, D-galactose. Individuals of blood group AB contain both A and B antigens on their red cells. Individuals of blood group O lack a functional transferase and hence do not transfer either sugar of type A or B. The distribution of the different ABO types differs substantially between different populations as illustrated in Figure 6.1. Prominent is...

Affinity Based Biosensors for Biomolecular Interaction Analysis

Affinity-based biosensors have in recent years found increasing application for monitoring molecular interactions. These instruments use evanescent wave detection technology, and measure interactions by immobilizing one of the interactants (the ligand or target molecule) on the surface of a sensor chip. A solution containing the other interactant (the analyte ) is then applied to the chip. Binding of analyte to the surface results in a response proportional to the mass bound. The method is general with respect to interactant identity and can be applied to all molecules expressing affinity for each other, such as biomolecules (proteins, nucleic acids, carbohydrates or lipids), low-molecular-weight compounds (signaling substances or pharmaceuticals), or larger particles (vesicles, viruses, bacteria, or cells). The identity and biological activity of the immobilized ligand determines the specificity of the interaction analysis. The interaction process is followed in real time without...

Hyperemesis gravidarum

Hyperemesis gravidarum occurs in the extreme 0.5 to 1 of patients who have intractable vomiting. Patients with hyperemesis have abnormal electrolytes, dehydration with high urine-specific gravity, ketosis and acetonuria, and untreated have weight loss 5 of body weight. Intravenous hydration is the first line of therapy for patients with severe nausea and vomiting. Administration of vitamin B1 supplements may be necessary to prevent Wernicke's encephalopathy.

Mechanisms of Renal Dysfunction

Oxidative stress is defined as an imbalance between prooxidants and antioxidants. Reactive oxygen species (ROS) are intermediary metabolites that are normally produced in the course of oxygen metabolism. There are many reactive oxygen species that are produced by all cell types and can have profound effects on the vascular system to impact blood pressure regulation. Oxidative stress increases during hypertension due to increased production of ROS such as superoxide, hydroxy radical, and hydrogen peroxide and or decreased superoxide dismutase (SOD), which scavenges ROS. Most recent attention has been given to the role of superoxide. There are many enzymatic sources of superoxide including nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, xanthine oxidase, nitric oxide synthase, and cytochrome P450. ROS can react with and denature proteins, lipids, nucleic acids, carbohydrates, and other molecules leading to inflammation, apoptosis, fibrosis, and cell proliferation. However,...

Incomplete Quaternary Structure

A second example is that of PDB entry 1buu which is shown in Figure 4. Again, the model as deposited in the PDB looks somewhat odd with a single, and quite long, helix protruding from the rest of the structure (Figure 4a). In this case the biological unit is a trimer (Figure 4b), with the protruding helices packing together to form a coiled-coil which is inserted into a cellular membrane, the remainder of the structure forming a lectin domain that recognizes carbohydrates.

Discovery of Correction In Vitro Uptake and Treatment of MPS I

Correct the metabolic defect behind the GAG accumulation 3 . Although they did not know initially that this factor was an enzyme, it was clear that it was a diffusible factor and that it was very potent at very low concentrations. By 1971, the identity of the corrective factor for MPS I was determined to be a-L-iduronidase 4 , which was modified in some manner that made it distinct on heparin chromatography from noncorrective enzyme 24 . In 1977, Kaplan, et al. 25 determined that the modification that allowed for a-L-iduroni-dase to be corrective was a mannose 6-phosphate moiety on the N-linked high-mannose carbohydrates. Even with both the identity of the enzyme and the reason for its corrective properties known, the ability to test ERT in MPS I was limited by the lack of available sources of appropriately modified enzyme. Enzyme from tissue sources lacked mannose 6-phosphate and was not corrective. In addition, the native enzyme was present in vanish-ingly small quantities in...

Size Exclusion Chromatography with OnLine Light Scattering

SEC and static light scattering are complimentary techniques to determine the molecular weight of proteins. Integrating the two techniques offers numerous advantages over using either one in isolation. While batch-mode light scattering determines overall average molecular weights for heterogeneous and associating or interacting protein samples, size fractionation by SEC helps determine molecular weights of individual species in a sample solution. In addition, although SEC is a simple and fast method for estimating the molecular weights of many proteins based on their elution position, there are several problems with this conventional SEC approach. One is that the elution position depends not only on the molecular weight of the protein, but also on its shape. A protein with extended structure elutes earlier than expected and gives a higher molecular weight. Another problem is that the elution position will change if the protein has any tendency to interact with the column matrix. In...

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