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PubMed1977

Formation of methionine from alpha-amino-n-butyric acid and 5'-methylthioadenosine in the rat.

Adult rats may utilize two metabolites of methionine for the biosynthesis of this essential amino acid. In separate experiments methionine, labeled with 14C or with 35S was observed in plasma and urine following the administration of [2-14C]-alpha-amino-n-butyric acid or [35S]-5'-methylthioadenosine by stomach tube. Although alpha-amino-n-butyric acid (ABA) or homoserine, alone or with dietary sodium sulfate, choline, and/or S-methylcysteine, was not utilized for growth, weight loss in weanling rats was decreased by dietary cysteine when fed as an additive to a basal methionine-free, cysteine-free, labile methyl-free, sulfur-free diet. Following the addition of 10 mg ABA and 28 mg 5'-methylthioadenosine/day to the basal diet, growth response was equivalent to that occurring in rats receiving 27 mg of methionine/day with the basal diet. The implications of these findings for adaptation to protein restriction and a discussion of equilibrium and steady state conditions related to the increase in methionine content in the blood are presented.

Adenosine↗
PubMed1993

Oat bran but not a beta-glucan-enriched oat fraction enhances butyrate production in the large intestine of pigs.

Digestibility of polysaccharides and other macronutrients and the metabolic response of the microflora in the large intestine to a low dietary fiber wheat flour diet and three enriched diets with equal amounts of added dietary fiber (oat bran, a beta-glucan-enriched oat fraction and insoluble oat residues) were studied in ileal-cannulated pigs. The digestibility of starch was high in the small intestine (98-100%). At this site of the gastrointestinal tract there was also a significant degradation of mixed linked beta (1 --> 3; 1 --> 4)-D-glucan (beta-glucan) (45-54%), whereas arabinoxylan was quantitatively recovered in ileal effluent. Type and amount of polysaccharides passing the ileal-cecal junction had little effect of the density of microorganism in the large intestine (approximately 10(10) viable counts/g digesta) but did have a high impact on the activity of the flora in colon as measured by the concentration of ATP in digesta. The relative proportion of butyrate in the short-chain fatty acids in the luminal contents of the large intestine was 6.6-8.4% when the low dietary fiber wheat flour diet was fed. However, when either oat bran or insoluble residues were included in the diet, the level was raised to 9.3-11.2%. No effect was seen after the addition of the beta-glucan-enriched fraction. This study showed that arabinoxylan and not beta-glucan in the cell walls of oat bran was responsible for the enhanced butyrate production of oat bran.

Animals↗
PubMed2001

Wheat aleurone flour increases cecal beta-glucuronidase activity and butyrate concentration and reduces colon adenoma burden in azoxymethane-treated rats.

Processed wheat aleurone flour (WAF) is a source of insoluble fermentable dietary fiber that comes from the outer layers of the wheat kernel. A study was designed to evaluate WAF, wheat bran (WB) and alpha-cellulose as the source of dietary fiber (5 g/100 g of diet) in a semipurified high fat (20 g/100 g of as 1:1 lard/sunflower seed oil) diet fed to male Sprague-Dawley rats in which intestinal tumors were induced using azoxymethane (AOM). WAF at 33 g/100 g of diet (WAF33) and WB at 16 g/100 g of diet (WB16) increased the weight of feces and produced significantly higher concentrations in the cecum of the short-chain fatty acid butyrate (P < 0.001) than did no fiber (NF) and WAF added at only 10 g/100 g (1.5 g of dietary fiber) (WAF10). Cecal and fecal pH were both significantly lower in the WAF33 and WB16 treatments relative to control and no fiber treatments (P < 0.001). The intestinal tumors in the rats were assessed at 6 mo after the study began, and the WAF33- or WB16-fed rats showed a trend (P = 0.06) with 43% fewer colon adenomas relative to control. There was a significant inverse relationship between ss-glucuronidase activity and colon adenomas in the rat colon (r2 = 0.37, P = 0.001). WAF fiber influenced some metabolic markers of fermentation in the colon in a manner similar to that of WB, which, independent of the bulking effect, was associated with a trend to reduced colon adenomas. Significantly increased cecal ss-glucuronidase activity and/or butyrate concentrations may have protective influences in this context by mechanisms not yet fully elucidated.

Adenoma↗
PubMed2005

Hyaluronic acid butyric esters in cancer therapy.

In this review we focus on a promising novel histone deacetylase (HDAC) inhibitor (HA-But) obtained by the esterification of butyric acid (BA), the smallest HDAC inhibitor, with hyaluronic acid (HA), the main constituent of the extracellular matrix which selectively recognizes a transmembrane receptor (CD44) overexpressed in most primary cancers and associated with tumor progression. In vitro, HA-But has proved to be 10-fold more effective than BA in inhibiting the proliferation of a panel of human cancer cell lines, representative of the most common human cancers, and, similar to BA, to regulate the expression of some cell cycle-related proteins, to induce growth arrest in the G1/G0 phase of the cell cycle and to increase histone acetylation. In vivo, HA-But treatment has demonstrated a marked potency in inhibiting primary tumor growth and lung metastases formation from murine Lewis lung carcinoma (LL3) as well as liver metastases formation from intrasplenic implantation of LL3 or B16-F10 murine melanoma cells. In particular, the effect of s.c. and i.p. treatment with HA-But on liver metastases resulted, respectively, in 87 and 100% metastases-free animals, and in a significant prolongation of the survival time compared to the control groups. The results suggest that the presence of the HA backbone does not interfere with the biological activity of butyric residues and that HA-But could represent a promising cell-targetable antineoplastic agent for the treatment of primary and metastatic tumors.

Animals↗
PubMed2004

Butyrate suppresses hypoxia-inducible factor-1 activity in intestinal epithelial cells under hypoxic conditions.

Interaction between the products of intestinal bacteria and the intestinal epithelial cells is a key event in understanding the biological, physiological, and pathological functions of the intestinal epithelium. Here, we examined the effect of butyrate, one of the major intestinal bacterial products, on hypoxia-inducible factor-1 (HIF-1) activity under hypoxic conditions in intestinal epithelial cells. HIF-1 activity was assessed by luciferase assay using cytoplasmic extracts of intestinal epithelial cells, Caco-2, and IEC-6 cells. These cells were transiently transfected with hypoxia response element (HRE)-luciferase reporter plasmids and cultured under hypoxic conditions in the presence or absence of sodium butyrate (NaB). The effect of NaB on HRE DNA binding activity in Caco-2 cells under hypoxic conditions was assessed by electrophoretic mobility shift assay. Expression of a hypoxia-responsive gene encoding intestinal trefoil factor (ITF) in Caco-2 cells after NaB treatment was assessed using reverse-transcription PCR. The barrier function of Caco-2 cells under hypoxic conditions was also evaluated by transepithelial electrical resistance measurement. NaB suppressed up-regulation of HIF-1 transcriptional activity under hypoxic conditions in Caco-2 and IEC-6 cells. In parallel, NaB reduced HRE DNA binding activity under the same conditions. Furthermore, NaB down-regulated enhanced transcription of ITF gene. Addition of NaB under hypoxic conditions delayed recovery of transepithelial electrical resistance of the monolayers after hypoxia-reoxygenation treatment. These findings indicate that NaB suppresses HIF-1 transcriptional activity on hypoxia-responsive genes by reducing the HRE DNA binding activity under hypoxic conditions in intestinal epithelial cells.

Animals↗
PubMed2002

Roseburia intestinalis sp. nov., a novel saccharolytic, butyrate-producing bacterium from human faeces.

Five strains of butyrate-producing, anaerobic, gram-positive bacteria were isolated from human faecal material. These strains were slightly curved rods that showed motility by means of multiple subterminal flagella. The DNA G + C content of the strains was 29-31 mol%. A detailed investigation of the phenotypic and phylogenetic characteristics of the strains revealed that they represent a novel species of anaerobic, low-G+C-content, butyrate-producing bacterium that shows net acetate utilization during growth on media containing carbohydrates and short-chain fatty acids. The 16S rRNA gene sequences of the five isolates were determined and they confirmed that these strains were closely related to each other. Phylogenetic analysis indicated that the most closely related species are Eubacterium rectale, Eubacterium oxidoreducens and Roseburia cecicola, members of cluster XIVa of the Clostridium subphylum of gram-positive bacteria, although they share less than 95% sequence identity with the novel strains. It is proposed that a novel species, Roseburia intestinalis sp. nov., be created, with strain L1-82T (= DSM 14610T = NCIMB 13810T) as the type strain.

Base Composition↗
PubMed1982

Control of interferon mRNA levels and interferon yields in butyrate and 5'-bromodeoxyuridine-treated Namalwa cells.

Treatment of Namalwa cells with butyrate or 5'-bromodeoxyuridine (BrdUrd) before induction with Sendai virus caused an increase in the production of both interferon (IFN) and interferon mRNA (IFN mRNA). However, the increase in IFN mRNA did not completely account for the increase in IFN yield. The treatments did not affect the time course of IFN mRNA transcription and translation, or the association of IFN mRNA with polysomes. Likewise, the treatments did not alter the post-translational fate of the IFN produced. We conclude that butyrate and BrdUrd affect IFN production at the level of transcription or processing of IFN mRNA and suggest that increased efficiency of translation provides an additional level of control.

Bromodeoxyuridine↗
PubMed1981

Presence of diaminopimelic acid in propionate-negative Bacteroides species and in some butyric acid-producing strains.

AThe presence of diaminopimelic acid (m-DAP) in strains of Bacteroides melaninogenicus, B. bivius and other species as well as in unidentified strains of Bacteroides was investigated by thin-layer chromatography. Strains of B. bivius and B. disiens all contained m-DAP as did the subspecies intermedius and melaninogenicus of B. melaninogenicus. Strains of B. asaccharolyticus and similar black pigment-producing butyrate-positive isolates showed heterogeneity. Asaccharolytic strains were DAP negative, whereas two strains fermenting glucose were positive. Some of the non-pigmented propionate-negative and butyrate-negative unidentified strains also contained DAP. The consistent finding of m-DAP in strains of B. bivius, B. disiens, and B. melaninogenicus indicates that DAP detection might be of value in the identification of these species.

Amino Acids, Diamino↗
PubMed1998

Identification and characterization of a monocarboxylate transporter (MCT1) in pig and human colon: its potential to transport L-lactate as well as butyrate.

1. Oligonucleotide primers based on the human heart monocarboxylate transporter (MCT1) cDNA sequence were used to isolate a 544 bp cDNA product from human colonic RNA by reverse transcription-polymerase chain reaction (RT-PCR). The sequence of the RT-PCR product was identical to that of human heart MCT1. Northern blot analysis using the RT-PCR product indicated the presence of a single transcript of 3.3 kb in mRNA isolated from both human and pig colonic tissues. Western blot analysis using an antibody to human MCT1 identified a specific protein with an apparent molecular mass of 40 kDa in purified and well-characterized human and pig colonic lumenal membrane vesicles (LMV). 2. Properties of the colonic lumenal membrane L-lactate transporter were studied by the uptake of L-[U-14C]lactate into human and pig colonic LMV. L-Lactate uptake was stimulated in the presence of an outward-directed anion gradient at an extravesicular pH of 5.5. Transport of L-lactate into anion-loaded colonic LMV appeared to be via a proton-activated, anion exchange mechanism. 3. L-Lactate uptake was inhibited by pyruvate, butyrate, propionate and acetate, but not by Cl- and SO4(2-). The uptake of L-lactate was inhibited by phloretin, mercurials and alpha-cyano-4-hydroxycinnamic acid (4-CHC), but not by the stilbene anion exchange inhibitors, 4,4'-diisothiocyanostilbene-2, 2'-disulphonic acid (DIDS) and 4-acetamido-4'-isothiocyanostilbene-2, 2'-disulphonic acid (SITS). 4. The results indicate the presence of a MCT1 protein on the lumenal membrane of the colon that is involved in the transport of L-lactate as well as butyrate across the colonic lumenal membrane. Western blot analysis showed that the abundance of this protein decreases in lumenal membrane fractions isolated from colonic carcinomas compared with that detected in the normal healthy colonic tissue.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2006

H2 and acetate transfers during xylan fermentation between a butyrate-producing xylanolytic species and hydrogenotrophic microorganisms from the human gut.

The aim of this work was to investigate in vitro interrelationships during xylan fermentation between an H2 and butyrate-producing xylanolytic species recently isolated in our laboratory from human faeces and identified as Roseburia intestinalis and the H2-utilizing acetogen Ruminococcus hydrogenotrophicus or the methanogen Methanobrevibacter smithii. H2 transfer between M. smithii or Ru. hydrogenotrophicus and the xylanolytic species was evidenced, confirming the great potential of these H2-consuming microorganisms to reutilize fermentative H2 during fibre fermentation in the gut. In addition, acetate transfer was demonstrated between the xylanolytic Roseburia sp. and the acetogenic species, both metabolites transfers leading to butyric fermentation of oat xylan without production of H2.

Acetates↗
PubMed1996

Pathways of acetate, propionate, and butyrate formation by the human fecal microbial flora.

The pathways of short-chain fatty acid (SCFA; acetate, propionate, and butyrate) formation from glucose were determined for the human fecal microbial communities of two subjects. The pathways were identified by radioisotope analysis of the SCFA and CO2 obtained after incubation of fecal suspensions with glucose under 20% CO2 with [1-14C]glucose, [3,4-14C]glucose, or 14CO2. Acetate was chemically degraded to learn the labeling of the methyl and carboxyl carbons. The labeling of CO2 and acetate showed that the major route of glucose catabolism was the Embden-Meyerhof-Parnas pathway, with production of CO2 from pyruvate carboxyl carbon. Labeling of the methyl and carboxyl carbons of acetate by 14CO2 or [3,4-14C]glucose proved that acetate was formed from CO2 by the Wood-Ljungdahl pathway. CO2 reduction accounted for about one-third of the acetate formed by suspensions from subject 1 and about one-fourth of the acetate formed by suspensions from subject 2. Propionate was formed by a CO2 fixation pathway, and butyrate was formed by classical routes of acetyl-S coenzyme A condensation. The amount of CO2 formed from [1-14C] glucose and acetate labeling patterns obtained with the other 14C precursors indicated that the Entner-Doudoroff, transketolase-transaldolase, and heterolactic pathways were not significant. Fermentation of cabbage cellulose by subject 1 followed the same pathways as were used for glucose. The results with suspensions from subject 2 suggested that some radioactive acetate was formed from the C-3 of glucose by the Bifidobacterium pathway.

Acetates↗
PubMed2000

Toxins, butyric acid, and other short-chain fatty acids are coordinately expressed and down-regulated by cysteine in Clostridium difficile.

It was recently found that a mixture of nine amino acids down-regulate Clostridium difficile toxin production when added to peptone yeast extract (PY) cultures of strain VPI 10463 (S. Karlsson, L. G. Burman, and T. Akerlund, Microbiology 145:1683-1693, 1999). In the present study, seven of these amino acids were found to exhibit a moderate suppression of toxin production, whereas proline and particularly cysteine had the greatest impact, on both reference strains (n = 6) and clinical isolates (n = 28) of C. difficile (>99% suppression by cysteine in the highest toxin-producing strain). Also, cysteine derivatives such as acetylcysteine, glutathione, and cystine effectively down-regulated toxin expression. An impact of both cysteine and cystine but not of thioglycolate on toxin yield indicated that toxin expression was not regulated by the oxidation-reduction potential. Several metabolic pathways, including butyric acid and butanol production, were coinduced with the toxins in PY and down-regulated by cysteine. The enzyme 3-hydroxybutyryl coenzyme A dehydrogenase, a key enzyme in solventogenesis in Clostridium acetobutylicum, was among the most up-regulated proteins during high toxin production. The addition of butyric acid to various growth media induced toxin production, whereas the addition of butanol had the opposite effect. The results indicate a coupling between specific metabolic processes and toxin expression in C. difficile and that certain amino acids can alter these pathways coordinately. We speculate that down-regulation of toxin production by the administration of such amino acids to the colon may become a novel approach to prophylaxis and therapy for C. difficile-associated diarrhea.

Amino Acid Sequence↗
PubMed1995

Interleukin 8 secretion by colonic crypt cells in vitro: response to injury suppressed by butyrate and enhanced in inflammatory bowel disease.

Epithelia from several sites exhibit inducible secretion of interleukin 8 (IL-8). This study aimed to characterise secretion of IL-8 by colonic epithelial cells in vitro. Colonic crypt cells were isolated enzymatically from resected colon and the IL-8 content of culture supernates was measured by ELISA. The rate of secretion of IL-8 accelerated and levels of IL-8 transcripts increased appreciably during culture. Exposure to tumour necrosis factor alpha (TNF alpha) failed to increase secretion further. Secretion was not induced by the enzymatic digestion or by serum used in the culture medium but was significantly inhibited by butyrate, by a mean of 23%. Control experiments indicated that colonic crypt cells were the likely source. The secretion of IL-8 over 24 hours by cells from uninflamed mucosa of patients with ulcerative colitis or Crohn's disease was more than twofold that from normal cells, while that from cancer bearing colons was normal. TNF alpha (10 mM) significantly suppressed IL-8 secretion only in the ulcerative colitis group and the change was different to those in the normal (p = 0.007) and Crohn's disease groups (p = 0.012). Cells from inflamed areas secreted more IL-8 than those from autologous uninflamed areas (p = 0.009) but responses to modulating factors were no different. The induction of IL-8 secretion by colonic crypt cells in vitro is probably a response to injury associated with isolation and culture. It is suppressed by butyrate and increased in inflammatory bowel disease independently of the presence of mucosal inflammation. Whether epithelial derived IL-8 plays a part in the pathogenesis of inflammatory bowel disease is not yet clear.

Adolescent↗
PubMed2003

Sodium butyrate inhibits cell growth and stimulates p21WAF1/CIP1 protein in human colonic adenocarcinoma cells independently of p53 status.

Butyric acid, one of the short-chain fatty acids produced by microbial fermentation in the colon, exhibits antiproliferative activities in various cancer cell lines. The initial objective of the study was to assess whether the effect of sodium butyrate (NaB) on cell growth differed by p53 status of the cells. Four human colorectal adenocarcinoma cell lines were used: HT29 (p53 point mutation), Caco2 (p53 truncation), LS513 (p53 wild type), and Lovo (p53 wild type). NaB significantly inhibited cell growth in all four cell lines. NaB arrested HT29 and LS513 cells in G0/G1 and Caco2 and Lovo in G2-phase. A second objective was to determine whether NaB similarly affected the cyclin-dependent kinase inhibitor, p21WAF1/CIP1. In all cell lines, p21 mRNA levels were immediately elevated after NaB exposure, and p21 protein levels were increased within 6 h. NaB increased p21 promoter activity in both Caco2 and Lovo, suggesting p53 independence. NaB did not influence p21 mRNA stability. Although three DNase I hypersensitivity sites were identified in the region of the p21 gene, induction of p21 mRNA by NaB was not accompanied by relaxation of the chromatin in the region of the p21 gene.

Adenocarcinoma↗
PubMed2002

Differential activation of the IGF binding protein-3 promoter by butyrate in prostate cancer cells.

Sodium butyrate (NaB), a dietary micronutrient, is a potent growth inhibitor that initiates cell differentiation in many cell types, including prostate cancer cells. The molecular mechanisms by which these effects occur remain largely unknown. In this study, we investigated the effects of NaB on the expression of IGF binding protein (IGFBP)-3, a known growth regulator, in two human prostate cancer cell lines (PC-3 and LNCaP). Treatment with NaB (0-10 mM) caused a dose-dependent stimulation of IGFBP-3 mRNA expression and parallel increases in protein levels. A specific histone deacetylase inhibitor, trichostatin A (TSA) similarly induced IGFBP-3 expression, indicating that histone hyperacetylation may be critical in the regulation of IGFBP-3 expression. To investigate the molecular mechanism of NaB-regulated IGFBP-3 expression, 1.87 kb of the human IGFBP-3 gene promoter was cloned into the pGL2-basic luciferase reporter vector. In both PC-3 and LNCaP cells, NaB (10 mM) significantly increased luciferase activity 20- to 30-fold, compared with the untreated control. However, using 5' sequential deletion constructs of the IGFBP-3 promoter, the NaB response sequences in the IGFBP-3 promoter were different in PC-3 and LNCaP cells. Our studies identified a region, -75 to +69 from the start of transcription (+1), that is fully inducible by NaB treatment in LNCaP cells, but not in PC-3 cells. Unlike other well characterized NaB-regulated genes, Sp1 DNA sequences are not involved in NaB up-regulation of IGFBP-3 gene in LNCaP cells. Further deletion studies identified two independent regions critical for NaB-induced transactivation in LNCaP cells. These regions contain consensus binding sites for p53 and GATA, respectively, but mutational analyses and gel shift assays suggested that, while the p53 response element is required for NaB responsiveness, neither p53 nor GATA are involved. In summary, we have demonstrated that 1) NaB significantly up-regulates IGFBP-3 mRNA and protein levels in PC-3 and LNCaP prostate cancer cells; and 2) novel butyrate- responsive elements lacking consensus Sp1 sites are used in LNCaP cells.

Blotting, Northern↗
PubMed1988

Requirement of serum pretreatment for induction of alkaline phosphatase activity with prednisolone, butyrate, dibutyryl cyclic adenosine monophosphate and NaCl in human liver cells continuously cultured in serum-free medium.

A cell line (HuL-1) derived from normal fetal human liver was adapted to grow continuously in a modified Eagle's minimum essential medium without serum or hormones. The population doubling time of this adapted cell line (HuL-1-317) was about 72 h and the modal number of chromosomes was 54. The morphology of HuL-1-317 cells was round in the absence of serum, but at 37 degrees C with the addition of serum (1-10%), the cells flattened. HuL-1-317 cells had a low level of alkaline phosphatase activity. However the enzyme activity was slightly enhanced by the combination of prednisolone, butyrate, dibutyryl cyclic adenosine monophosphate and a hypertonic concentration of NaCl after 3 days of incubation at 37 degrees C. The increase in alkaline phosphatase activity with the four agents was further amplified dose-dependently by the pretreatment of the cells with serum. The stimulatory effect of the serum was evident at concentrations as low as 1%, and was maximal at 20%. The half life of the effect of serum on alkaline phosphatase induction was 48 h at 37 degrees C. Serum alone could not enhance the enzyme activity without the four agents. The present results indicate that serum contributes to the regulation of alkaline phosphatase induction by the combination of prednisolone, butyrate, dibutyryl cyclic adenosine monophosphate and NaCl in fetal human liver cells (HuL-1-317).

Alkaline Phosphatase↗
PubMed2004

High-productivity continuous biofilm reactor for butanol production: effect of acetate, butyrate, and corn steep liquor on bioreactor performance.

Corn steep liquor (CSL), a byproduct of the corn wet-milling process, was used in an immobilized cell continuous biofilm reactor to replace the expensive P2 medium ingredients. The use of CSL resulted in the production of 6.29 g/L of total acetone-butanol-ethanol (ABE) as compared with 6.86 g/L in a control experiment. These studies were performed at a dilution rate of 0.32 h-1. The productivities in the control and CSL experiment were 2.19 and 2.01 g/(L.h), respectively. Although the use of CSL resulted in a 10% decrease in productivity, it is viewed that its application would be economical compared to P2 medium. Hence, CSL may be used to replace the P2 medium. It was also demonstrated that inclusion of butyrate into the feed was beneficial to the butanol fermentation. A control experiment produced 4.77 g/L of total ABE, and the experiment with supplemented sodium butyrate produced 5.70 g/L of total ABE. The butanol concentration increased from 3.14 to 4.04 g/L. Inclusion of acetate in the feed medium of the immobilized cell biofilm reactor was not found to be beneficial for the ABE fermentation, as reported for the batch ABE fermentation.

Acetates↗
PubMed1980

Differentiative and proliferative effects of (But)2cAMP, n-butyric acid and prednisolone on the malignant melanoma cell line (TM-1) in vitro and in vivo.

Both (But)2cAMP and n-butyric acid suppressed the proliferation of TM-1 cells and induced the formation and elongation of the cellular processes in vitro. Prednisolone stimulated the proliferation of TM-1 cells and increased the number of polygonal cells in vitro. These results indicated that (But)2cAMP and n-butyric acid induce the differentiation of TM 1 cells, whereas prednisolone brings the cells in an undifferentiated state. The growth of TM-1 cell tumor transplanted into nude mice was suppressed by 5 mg of (But)2cAMP and 1 mg of theophylline. Remarkable histological changes were seen in the tumors treated with these agents as compared with untreated tumors. This suggests that (But)2cAMP suppresses the proliferation of TM-1 cells in vivo.

Animals↗