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Fatty-acid biosynthesis in a branched-chain alpha-keto acid dehydrogenase mutant of Streptomyces avermitilis.

Fatty-acid biosynthesis by a branched-chain alpha-keto acid dehydrogenase (bkd) mutant of Streptomyces avermitilis was analyzed. This mutant is unable to produce the appropriate precursors of branched-chain fatty acid (BCFA) biosynthesis, but unlike the comparable Bacillus subtilis mutant, was shown not to have an obligate growth requirement for these precursors. The bkd mutant produced only straight-chain fatty acids (SCFAs) with membrane fluidity provided entirely by unsaturated fatty acids (UFAs), the levels of which increased dramatically compared to the wild-type strain. The levels of UFAs increased in both the wild-type and bkd mutant strains as the growth temperature was lowered from 37 degrees C to 24 degrees C, suggesting that a regulatory mechanism exists to alter the proportion of UFAs in response either to a loss of BCFA biosynthesis, or a decreased growth temperature. No evidence of a regulatory mechanism for BCFAs was observed, as the types of these fatty acids, which contribute significantly to membrane fluidity, did not alter when the wild-type S. avermitilis was grown at different temperatures. The principal UFA produced by S. avermitilis was shown to be delta 9-hexadecenoate, the same fatty acid produced by Escherichia coli. This observation, and the inability of S. avermitilis to convert exogenous labeled palmitate to the corresponding UFA, was shown to be consistent with an anaerobic pathway for UFA biosynthesis. Incorporation studies with the S. avermitilis bkd mutant demonstrated that the fatty acid synthase has a remarkably broad substrate specificity and is able to process a wide range of exogenous branched chain carboxylic acids into unusual BCFAs.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗

Metabolic effects of D-psicose in rats: studies on faecal and urinary excretion and caecal fermentation.

D-psicose (D -ribo-2-hexulose), a C-3 epimer of D-fructose, is one of the "rare sugars" present in small quantities in commercial carbohydrate complex or agricultural products. We investigated the absorption and excretion of D-psicose when orally administrated (5g/kg body weight) to Wistar rats, and the fermentation of D-psicose was measured as caecal short-chain fatty acids (SCFAs) when fed to rats in controlled diets (0, 10, 20 and 30%). Urinary and faecal excretions of D-psicose over the 24 h, following a single oral administration, were 11-15% of dosage for the former and 8-13% of dosage for the latter. Serum D-psicose concentration and D-psicose in the contents of stomach and small intestines decreased progressively after administration. D-psicose in caecum contents was detected after 3h and 7h administration, but not after 1h. Rats fed on D-psicose diets showed short-chain fatty acid production with caecal hypertrophy. These results suggest that D-psicose is partly absorbable in the digestive tract and is excreted into urine and faeces. As with other poorly absorbed dietary carbohydrates, D-psicose is fermented in the caecum by intestinal microflora.

Administration, Oral↗

Intestinal fuels: glutamine, short-chain fatty acids, and dietary fiber.

In recent years, considerable research has focused on the physiologic effects and clinical uses of three dietary constituents thought to be trophic to the intestinal tract in human beings: glutamine, short-chain fatty acids (SCFAs), and dietary fiber. Glutamine is an important nitrogen-carrying amino acid that may be "conditionally essential" in certain disease states to support the gut barrier and immune function and overall protein use. Colonic irrigations with SCFA preparations have demonstrated enhanced healing of bowel tissue in animals and human beings. Dietary fiber supports bacterial SCFA production, normal stool output, and the gut barrier and immune function. However, optimal fiber doses for various medical conditions are not known, and the risk for gastrointestinal (GI) obstruction, diarrhea, gas, and bloating necessitates careful selection of patients and daily monitoring of fiber tolerance. A review of the current literature indicates that widespread use of glutamine and SCFA additives parenterally and enterally awaits further evidence of safety and efficacy in human beings, establishment of appropriate doses, and advances in formulation technology. Administration of dietary fiber to enhance bowel motility should be considered in long-term tube-fed patients with intact GI function and sufficient fluid tolerance to permit hydration of fiber. Industrywide agreement on fiber analysis methods and labeling standards (eg, fiber fermentability vs solubility) would facilitate selection of enteral products. To streamline studies and optimize research efforts in future clinical trials, standard criteria for evaluating GI function, diarrheagenic factors, and intestinal outcome variables should be established.

Animals↗

Validity of repeated dietary measurements in a dietary intervention study.

The aim of the study was to evaluate the compliance in a dietary intervention study. When drawing conclusions about the relationship between dietary intake and disease occurrence/disease-related variables it is important to obtain valid dietary data. 20 healthy, non-smoking normal-weight omnivores changed from a mixed to a lactovegetarian diet. Dietary surveys (four 24 h recalls per person and time-period), urinary and faecal sample collections were performed before and 3, 6 and 12 months after the dietary shift. The validation of energy, protein, sodium and potassium yielded approximately the same ratio of dietary intake to biological marker at 0 and 3 months. This ratio decreased towards 6 months and continued to decrease towards 12 months. The fibre intake was compared to the total faecal weight directly and indirectly by calculating the fibre intake from the stool weight, the water content in faeces and the excretion of short-chain fatty acids (SCFAs). These four methods of fibre validation showed that the ratio of dietary intake to biological marker was always highest at 12 months, indicating an overestimation of the fibre intake at the end of the study. This is the first time these methods of validating fibre intake have been used in an epidemiological study. The ratio of dietary calcium intake to urinary and faecal calcium excretion did not show any statistical difference between the period before and 3 months after the dietary shift. To conclude, almost all investigated dietary data show approximately the same validity before and 3 months after the dietary shift, and show the least validity 12 months after the dietary shift. Thus, this study demonstrates that it is difficult to obtain valid dietary data 1 year after a drastic dietary change, indicating a decreased compliance to the new dietary regimen at the end of the 1 year study period. This represents important information when attempting to relate biological effects to dietary intake, and illustrates the importance of using biological markers for food intake in dietary surveys.

Adult↗

Resistant starch and colorectal neoplasia.

There are several approaches to examining the relationship between resistant starch (RS) and development of colorectal cancer (CRC). These include examination of epidemiological relationships, objective testing of effects of RS given to humans on biological events of relevance to CRC, and studies in animal models where protection and mechanisms of protection can be directly tested. Nine epidemiological studies have examined the relationship between starch and CRC and/or adenomas. Most show a significant protective effect. However, epidemiological tools for measuring consumption of RS are poorly developed and so a benefit for RS can only be inferred. On balance, the magnitude of protection by starch appears to be in the order of 25-50%. Human intervention studies have examined the effect of various types and amounts of RS consumption on colonic biology. To generalize from these studies, RS softens stools and increases stool bulk, decreases pH, increases short-chain fatty acids (SCFAs) including butyrate, reduces products of protein fermentation, and decreases bile salts in fecal water. Such changes seem to be achieved within about 4 weeks of commencing consumption. The greatest effects are seen with the highest doses where increased fecal starch recovery is observed. A modest number of animal studies have been undertaken. Those examining effects of RS on colonic biology and biomarkers for CRC confirm and extend the results in humans. RS modifies the lumenal environment, largely through altered fermentation of polysaccharides and proteins. RS also affects epithelial biology in that it increases apoptotic deletion of genetically damaged cells. More work is needed to define what types and combinations of RS, perhaps with probiotics, exert the greatest effects on colonic environment and epithelial biology, and then to test these in the cancer models for their protective effect. A few studies have examined effect of RS on cancer as an end point in several rodent models, but the results are not clear cut. In conclusion, consumption of RS dramatically affects the colonic lumenal environment and facilitates apoptotic deletion of genetically damaged cells in the colon, several of which are considered to be biomarkers associated with risk for CRC. These effects can be interpreted as reflecting improved colonic health, which might be of benefit in protection against CRC. Direct evidence for protection is still not available.

Animals↗

The role of short-chain fatty acid metabolism in colonic disorders.

During the past decade it has become evident that colonic mucosal metabolism is more complex than previously suspected. Luminal short-chain fatty acids (SCFAs) are recognized as an essential fuel source for colonocytes, particularly in the distal colon. Their absence may explain the development of diversion colitis; however, this has not been confirmed by clinical trials. The histologic, endoscopic, and metabolic similarities between diversion colitis and ulcerative colitis suggest that a nutritional SCFA deficiency state may play a role in the pathogenesis of these disorders. Diversion colitis and continent urinary diversion, utilizing distal and proximal colon reservoirs, provide in vivo models to study normal colonic mucosa in circumstances of reduced intraluminal SCFA concentrations and altered luminal effluent. Further studies utilizing these models would enhance our understanding of the regional differences in mucosal cell metabolism and adaptability and, hopefully, provide therapeutic alternatives for the management of colonic disorders. The welfare of colonic mucosa, as it relates to SCFA metabolism, awaits another exciting decade of investigation.

Colonic Diseases↗

Digestive processes in the human colon.

The presence or absence of a normal small bowel is evidently important for the digestion and absorption of nutrients in humans. The importance, however, of the large intestine as an organ with digestive potential and an ability to salvage energy is much less appreciated. Whereas the bacterial fermentation of plant polysaccharides, with the production and absorption of short-chain fatty acids (SCFAs), contributes 60-90% of all the energy requirements in plant-eating animals, the colonic fermentation in humans is of minor importance for nutrition, with only 5-10% of the energy requirements available from colonic digestion of starch, nonstarch polysaccharides, and protein not absorbed in the small bowel, if the intestine has a normal length and function. In contrast, the digestive resource of the large bowel might be important for people with reduced upper-gut function and with the malabsorption of large amounts of dietary nutrients to the cecum, e.g., in patients with short bowel. The substrates available for bacterial fermentation and for the maintenance of the colonic flora are largely starch and nonstarch polysaccharides (dietary fiber). Whereas nonstarch polysaccharides are undegradable by amylase in the small intestine, starch is hydrolyzed by amylase, but with very different rates and to very different degrees dependent on the origin and structure of the starches. The unequal susceptibility to amylase explains the different amount of starch that is not absorbed in the small intestine. The rate of colonic fermentation of starch has also been shown to relate to the rate of starch hydrolysis by amylase.(ABSTRACT TRUNCATED AT 250 WORDS)

Colon↗