Factors Affecting Energy Metabolism and Prognosis in Patients with Amyotrophic Lateral Sclerosis.

BACKGROUND/AIMS: Nutritional status is a factor affecting prognosis in patients with amyotrophic lateral sclerosis (ALS). Here, we aimed to clarify the factors associated with hypermetabolism and the prognosticators of ALS. METHODS: Forty-two inpatients (22 men, 20 women) diagnosed with ALS according to the revised El-Escorial criteria were investigated. The following data were retrospectively analyzed: anthropometric measurements, blood biochemistry, disease severity, basal energy expenditure (BEE), resting energy expenditure (REE) measured by indirect calorimetry, spirometry, and bioelectrical impedance analysis. Single and multiple regression analysis was performed to examine factors affecting REE and metabolic changes (defined as the ratio of REE to fat-free mass [FFM]). The Kaplan-Meier method was used to examine factors associated with the occurrence of cumulative events (death or tracheostomy). RESULTS: Among the 42 inpatients, REE was significantly higher than BEE, indicating hypermetabolism in ALS. Multiple regression analysis revealed that REE/FFM is strongly associated with the skeletal muscle index (-3.746 to -1.532, p < 0.0001) and percent forced vital capacity (%FVC) (-0.172 to -0.021, p = 0.013). Moreover, both the skeletal muscle index and %FVC were significant prognosticators associated with the occurrence of cumulative events. CONCLUSIONS: Energy metabolism was elevated in ALS, and respiratory status and muscle mass were associated with the hypermetabolism and poor prognosis. Adequate nutritional support may improve outcomes in ALS by preventing deterioration of respiratory status and reduction in muscle mass.

Factors affecting pancreatic hyperamylasemia in patients undergoing peroral single-balloon enteroscopy.

BACKGROUND AND AIM: Acute pancreatitis following balloon-assisted enteroscopy is a rare but serious complication. The causative mechanism is uncertain and prevention strategies are not established. We conducted a retrospective study to clarify the risk factors for pancreatic hyperamylasemia. METHODS: Eighty-four patients undergoing peroral single-balloon enteroscopy (SBE) were enrolled in this study. Serum pancreatic and salivary amylase levels were measured 2 h after endoscopic examination. RESULTS: We experienced three patients with post-SBE pancreatitis. Factors predicting pancreatic hyperamylasemia were: (i) elderly patients; (ii) deeper insertion; and (iii) clockwise insertion. In contrast, younger age at examination was a significant factor observed in salivary hyperamylasemia. CONCLUSIONS: It is important to measure pancreatic amylase and not total amylase after SBE. When carrying out peroral SBE, the distance of insertion should be reduced especially if the scope traces a clockwise loop or the subject is elderly.

Phase I clinical trial of the vaccination for the patients with metastatic melanoma using gp100-derived epitope peptide restricted to HLA-A*2402.

BACKGROUND: The tumor associated antigen (TAA) gp100 was one of the first identified and has been used in clinical trials to treat melanoma patients. However, the gp100 epitope peptide restricted to HLA-A*2402 has not been extensively examined clinically due to the ethnic variations. Since it is the most common HLA Class I allele in the Japanese population, we performed a phase I clinical trial of cancer vaccination using the HLA-A*2402 gp100 peptide to treat patients with metastatic melanoma. METHODS: The phase I clinical protocol to test a HLA-A*2402 gp100 peptide-based cancer vaccine was designed to evaluate safety as the primary endpoint and was approved by The University of Tokyo Institutional Review Board. Information related to the immunologic and antitumor responses were also collected as secondary endpoints. Patients that were HLA-A*2402 positive with stage IV melanoma were enrolled according to the criteria set by the protocol and immunized with a vaccine consisting of epitope peptide (VYFFLPDHL, gp100-in4) emulsified with incomplete Freund's adjuvant (IFA) for the total of 4 times with two week intervals. Prior to each vaccination, peripheral blood mononuclear cells (PBMCs) were separated from the blood and stored at -80°C. The stored PBMCs were thawed and examined for the frequency of the peptide specific T lymphocytes by IFN-γ- ELISPOT and MHC-Dextramer assays. RESULTS: No related adverse events greater than grade I were observed in the six patients enrolled in this study. No clinical responses were observed in the enrolled patients although vitiligo was observed after the vaccination in two patients. Promotion of peptide specific immune responses was observed in four patients with ELISPOT assay. Furthermore, a significant increase of CD8+ gp100-in4+ CTLs was observed in all patients using the MHC-Dextramer assay. Cytotoxic T lymphocytes (CTLs) clones specific to gp100-in4 were successfully established from the PBMC of some patients and these CTL clones were capable of lysing the melanoma cell line, 888 mel, which endogenously expresses HLA-restricted gp100-in4. CONCLUSION: Our results suggest this HLA-restricted gp100-in4 peptide vaccination protocol was well-tolerated and can induce antigen-specific T-cell responses in multiple patients. Although no objective anti-tumor effects were observed, the effectiveness of this approach can be enhanced with the appropriate modifications.

The sky blue method as a screening test to detect misplacement of percutaneous endoscopic gastrostomy tube at exchange.

BACKGROUND: During tube exchange for percutaneous endoscopic gastrostomy (PEG), a misplaced tube can cause peritonitis and death. Thus, endoscopic or radiologic observation is required at tube exchange to make sure the tube is placed correctly. However, these procedures cost extensive time and money to perform in all patients at the time of tube exchange. Therefore, we developed the "sky blue method" as a screening test to detect misplacement of the PEG tube during tube exchange. METHODS: First, sky blue solution consisting of indigocarmine diluted with saline was injected into the gastric space via the old PEG tube just before the tube exchange. Next, the tube was exchanged using a standard method. Then, we checked whether the sky blue solution could be collected through the new tube or not. Finally, we confirmed correct placement of the tube by endoscopic or radiologic observation for all patients. RESULTS: A total of 961 patients were enrolled. Each tube exchange took 1 to 3 minutes, and there were no adverse effects. Four patients experienced a misplaced tube, all of which were detectable with the sky blue method. Diagnostic parameters of the sky blue method were as follows: sensitivity, 94% (95%CI: 92-95%); specificity, 100% (95%CI: 40-100%); positive predictive value, 100% (95%CI: 100-100%); negative predictive value, 6% (95%CI: 2-16%). CONCLUSION: These results suggest that the number of endoscopic or radiologic observations to confirm correct replacement of the PEG tube may be reduced to one fifteenth using the sky blue method.

Milk fat globule epidermal growth factor-8 blockade triggers tumor destruction through coordinated cell-autonomous and immune-mediated mechanisms.

Carcinogenesis reflects the dynamic interplay of transformed cells and normal host elements, but cancer treatments typically target each compartment separately. Within the tumor microenvironment, the secreted protein milk fat globule epidermal growth factor-8 (MFG-E8) stimulates disease progression through coordinated alpha(v)beta(3) integrin signaling in tumor and host cells. MFG-E8 enhances tumor cell survival, invasion, and angiogenesis, and contributes to local immune suppression. We show that systemic MFG-E8 blockade cooperates with cytotoxic chemotherapy, molecularly targeted therapy, and radiation therapy to induce destruction of various types of established mouse tumors. The combination treatments evoke extensive tumor cell apoptosis that is coupled to efficient dendritic cell cross-presentation of dying tumor cells. This linkage engenders potent antitumor effector T cells but inhibits FoxP3(+) T reg cells, thereby achieving long-term protective immunity. Collectively, these findings suggest that systemic MFG-E8 blockade might intensify the antitumor activities of existing therapeutic regimens through coordinated cell-autonomous and immune-mediated mechanisms.

Similarities between N-glycan glycoform of tonsillar IgA1 and that of aberrant IgA1 abundant in IgA nephropathy patient serum.

BACKGROUND: There are many reports on the presence of an incompletely glycosylated O-linked oligosaccharide(s) in the IgA1 hinge region of some IgA nephropathy patients. As the candidates of such IgA1, tonsillar IgA1 and aberrant IgA1, which are abundant in an IgA nephropathy patient, were proposed. On the other hand, in mice, the abnormality of the N-linked oligosaccharide chain of IgA induced the IgA nephropathy. Therefore, analyses of the N-glycan glycoform on serum IgA1, aberrant IgA1 and tonsillar IgA1 were carried out using the 3-dimensional mapping method. RESULTS: The sugar chain composition was almost the same in these 3 IgA1 preparations. However, the structural characteristics for the aberrant IgA1 showed a drastic increase in the neutral N-glycans; in particular, 25% of the sugar chains in the aberrant IgA1 were the high mannose-type as compared with approximately 5%-6% in the serum IgA1 and tonsillar IgA1. The neutral complex-type N-glycan chain with fucose was higher in both the aberrant IgA1 and tonsillar IgA1 than in the serum IgA1. A typical component in the aberrant IgA1 was the fully galactosylated biantenna with the fucose residue. CONCLUSIONS: We found an abnormality in the N-linked oligosaccharides of the aberrant IgA1. In addition to our previous report about the abundance of asialo-O-linked oligosaccharide in both the tonsillar IgA and aberrant IgA, our results concerning the N-glycan glycoform of the aberrant IgA showed the possible promotion of its self-aggregation and its glomerular deposition by the synergistic difference in the O- and N-linked carbohydrate chains, and also the derivation of the aberrant IgA1 in the sera from the tonsillar tissue.

Effect of geranylgeranylacetone on gentamycin ototoxicity in rat cochlea culture.

OBJECTIVE: Geranylgeranylacetone (GGA), an anti-ulcer drug, has been reported to induce heat shock proteins (HSPs) in several animal organs. Purpose of this study was to investigate whether GGA has protective effect on gentamycin (GM) ototoxicity and whether it induces HSP70 in rat cochlea. METHODS: We used cochlea explant culture from postnatal 5-day rat. Explants were pre-incubated with GGA, then incubated with GM and GGA. The number of surviving outer hair cells (OHCs) labeled by phalloidin was counted to evaluate the protective effect of GGA. The expression of HSP70 in whole cochlea tissue was investigated by immunoblot analysis. RESULTS: The number of surviving OHCs was significantly high in GGA 10(-5)M group compared to GM only group and GGA 10(-6)M group. In the immunoblot analysis, HSP70 levels in GGA added groups were slightly high compared to simple culture group, but much lower than those in heat shock group. CONCLUSION: It was suggested that GGA-induced HSP70, and had partial protective effects on GM ototoxicity in the cochlea. GGA has possibility to be safe and useful treatment drug for cochlea disorder.

Disordered balance of IgA subclass production in the tonsils of some IgA nephropathy patients.

BACKGROUND: There are reports concerning the relationship between tonsillectomy and immunoglobulin A nephropathy (IgAN). Two reports on the biochemical analysis of over-produced IgA1 from IgAN patients were recently published. On the other hand, histochemical analysis of tonsillar tissue indicated the disordered balance in IgG and IgA producing cells and in the IgA subclass producing cells in IgAN patients. METHODS: IgA in tonsillar extracts and serum was separated into passed fraction (IgA2) and bound fraction (IgA1) by a jacalin-agarose column. Isoelectric focusing (IEF) analysis was carried out using an IPGphor instrument. The IgA content in each sample was analyzed by enzyme-linked immunosorbent assay (ELISA). RESULTS: The IgA1/IgA2 ratio of the tonsillar extracts from controls and IgAN patients was compared. The ratio distribution indicated statistically significant differences. The mean ratio for the control tonsil was 61/39. However, the ratio from eight out of thirty-two IgAN patients exhibited a higher value than the mean + 2SD (standard deviation) of the controls. Among them, three patients exhibited 92/8. Meanwhile, the ratios for serum by this method were close to the previously reported 89/11. There were no differences in the IgA1 IEF profile between the representative lowand high-IgA1 producing patients. CONCLUSIONS: This is the first report concerning IgA subclass distribution in tonsillar tissue. The ratio 61/39 for tonsillar IgA differ from the value (>90% of IgA1) in the previous histochemical report. The value is similar to the previous report for colostrum, whole saliva, jejunal fluid and bronchial fluid. The IgA1/IgA2 ratio distribution in the tonsillar extracts from the patients with chronic tonsillitis is significantly different from that of the IgAN patients.

Three-dimensional reconstruction of the human semicircular canals and measurement of each membranous canal plane defined by Reid's stereotactic coordinates.

OBJECTIVES: Equations for estimating the planar relationships of the human semicircular canals were devised by Blanks et al from a dissected bony labyrinth in a human skull. However, a similar study on the membranous semicircular canal planes has never been published. METHODS: In this study, the angle between each membranous canal plane and Reid's stereotactic horizontal plane was measured on serial histologic sections of 7 temporal bones from Japanese adults. We reconstructed the 3 semicircular canals by computer-aided 3-dimensional analysis. The angles between each pair of both bony and membranous canal planes were measured. RESULTS: In the bony labyrinth, the angles between the 2 canal planes of the lateral-anterior, anterior-posterior, and lateral-posterior pairs were 90.51 degrees +/- 2.98 degrees (mean +/- SD), 91.70 degrees +/- 1.85 degrees, and 94.52 degrees +/- 3.32 degrees, respectively. The angles between the 2 membranous canal planes of the lateral-anterior, anterior-posterior, and lateral-posterior pairs were 90.05 degrees +/- 4.74 degrees, 91.03 degrees +/- 2.93 degrees, and 91.92 degrees +/- 5.22 degrees, respectively. CONCLUSIONS: The data from our study of the membranous labyrinth showed that the angles between each canal plane and the others were much closer to 90 degrees than was found by Blanks et al for the bony labyrinth.

Compositional similarity between immunoglobulins binding to asialo-, agalacto-IgA1-Sepharose and those deposited in glomeruli in IgA nephropathy.

BACKGROUND: It has been found that the immunoglobulin A1-binding protein (IgA1-BP) can be separated from human serum using an asialo-, agalacto-IgA1 (aglyco-IgA1)-Sepharose column (13). As the IgA content in IgA1-BP was significantly higher in IgA nephropathy patients than that in other nephropathy patients, the relationship between IgA in IgA1-BP and glomerular deposited IgA was predicted. METHODS: IgA1-BP was separated from serum using the aglyco-IgA1-Sepharose column and the aglyco-IgA1-HPLC column. A jacalin-agarose column fractionated the IgA. The immunoglobulins were analyzed by the enzyme-linked immunosorbent assay (ELISA). RESULTS: A rapid separation method of IgA1-BP from serum by the aglyco-IgA1-HPLC column was developed and this column confirmed the reproduction of the IgA1-BP separation. The following similarities between IgA in IgA1-BP and glomerular deposited IgA were detected. A major portion of IgA in IgA1-BP was the IgA1 subclass. The IgA was rich in medium-size IgA and in the jacalin-high-affinity IgA1 fraction. The IgA showed a statistically significant lower kappa/lambda ratio in its light-chain composition than that of serum IgA, i.e. abundance in IgA bearing the lambda light chain. Other immunoglobulin classes (IgG and IgM) in IgA1-BP also exhibited a significantly low kappa/lambda ratio. CONCLUSIONS: In this experiment, preferential bindings of the IgA1 subclass, the medium-size IgA and the IgA with the lambda light chain to the aglyco-IgA1-column were observed. Based on previous reports concerning aglyco-IgA1 self-aggregation, the interaction of aglyco-IgA1 with matrix proteins and the rat glomerular deposition of artificially deglycosylated IgA1, IgA in IgA1-BP is thought to be partially the same molecule as the IgA deposited on the mesangial matrix in the IgA nephropathy patient.

Natriuretic peptides up-regulate aquaporin 3 in a human colonic epithelial cell line.

Several specialized channels termed aquaporins (AQPs) facilitate water transport in the gastrointestinal tract. AQP3 localizes to epithelial cells in the human small intestine and colon. However, the regulatory mechanisms responsible for AQP3 function in the gastrointestinal tract are not well understood. To characterize the regulation of AQP3 expression by atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP), we studied mRNA expression by Northern blotting, protein expression by Western blotting and DNA binding activity by electrophoretic mobility shift assay (EMSA) in the human colonic epithelial cell line HT-29. We also used several inhibitors to investigate signal transduction. AQP3 mRNA was up-regulated in addition to ANP (>100 nM) and BNP (>10 nM). The expression of AQP3 protein was enhanced at 1 h after the addition of ANP and BNP. The combination of protein kinase-A (PK-A) and protein kinase-G (PK-G) inhibitors completely inhibited the expression of AQP3 mRNA enhanced by ANP or BNP to its basal level. The EMSA of the cyclic-AMP response element (CRE) in HT-29 cells revealed a single band. These results indicate that ANP and BNP up-regulated the expression of AQP3 mRNA and protein, and both PK-A and PK-G dependent pathways mediated this effect.

Alteration of aquaporin mRNA expression after small bowel resection in the rat residual ileum and colon.

BACKGROUND: Diarrhea occurring after small bowel resection gradually improves due to intestinal adaptation. It is known that several water channels, termed aquaporins (AQP), are expressed in the gastrointestinal tract and facilitate water transport. However, the changes of AQP after bowel resection remain unclear. In the present paper, the alterations in AQP mRNA expression were investigated after a massive small bowel resection in the rat residual ileum and colon. METHODS: The 6-week-old male Sprague-Dawley rats (n = 15) underwent an 80% distal small bowel resection. The residual ileum and colon were dissected on postoperative day 1, 3, 5 or 7 (n= 3 on each day). Total RNA was purified from each mucosa, and the expressions of AQP and sodium-dependent glucose transporter (SGLT1) mRNA were analyzed by northern blot. The plasma vasoactive intestinal polypeptide (VIP) concentrations on the preoperative day and postoperative day 1 were assayed. RESULTS: In the residual small intestine, the expression of AQP-1 and AQP-3 mRNA increased significantly on postoperative day 1. The AQP-7 mRNA increased on postoperative day 3, but the AQP-4 mRNA did not change after the bowel resection. The SGLT1 mRNA gradually decreased after the bowel resection. In the colon, the expression of AQP-3 increased on postoperative day 1 and 7, but AQP-4 mRNA did not change after surgery. The AQP-8 mRNA levels increased slightly on postoperative day 7. Plasma VIP concentration did not change between preoperative day and postoperative day 1. CONCLUSIONS: These results indicate that several AQP, except for AQP-4, were up-regulated after a massive small bowel resection, and that AQP might play important roles during adaptation.

Tonsillar IgA1 as a possible source of hypoglycosylated IgA1 in the serum of IgA nephropathy patients.

BACKGROUND: There are many reports of incompletely glycosylated O-linked oligosaccharides on the IgA1 hinge region in certain IgA nephropathy patients. In addition, other reports have noted a relationship between tonsillectomy and IgA nephropathy. METHODS: Immunoglobulins from extracts of tonsillectomized tissue and other sources were analysed by isoelectric focusing (IEF) and by enzyme-linked immunosorbent assay (ELISA). RESULTS: The IEF profile of tonsillar IgA differed from that of serum IgA and it was enriched in cationic IgA. However, extracts from tonsillitis controls and IgA nephropathy patients exhibited profiles that were very similar. Enzymatic removal of sialic acid induced a shift of the peaks to the cathode side. The profiles of IgA from treated tonsillar extract and treated serum were closely overlapped. In addition, asialo Galbeta1,3GalNAc was clearly present in cationic IgA from tonsillar extract and in aberrant IgA1 from serum following enzymatic transfer of sialic acid to IgA1. Serum IgA also contained partly sialylated IgA1. Quantitative analysis of IgA and IgG in the extracts indicated that IgA was significantly higher, whereas IgG was significantly lower in IgA nephropathy patients. CONCLUSIONS: We found that the IgA1 produced in tonsillar tissue differed from serum IgA1. Furthermore, an overproduction of asialo IgA1 resulted from the disordered balance between IgA- and IgG-producing cells in the tonsils from the IgA nephropathy patient. Although it is unclear how such asialo IgA1 molecules are transferred from tonsil tissue to serum, a tonsillar source may produce a few micrograms of aberrant IgA1 that then appears in serum.

Mucosal permeability regulates receptor binding of luminal epidermal growth factor in the adult rat intestine.

Epidermal growth factor (EGF) stimulates repair in the damaged intestine, but its role in the normal intestine is not clear. Because EGF receptors are found on the basolateral surface but not the luminal surface, we hypothesized that mucosal permeability regulates EGF binding. Adult male rats were divided into 3 groups, one that was fed normal chow (the control), one that was starved for 4 days, and one that was given methotrexate (MTX) intragastrically (10 mg/kg/day for 3 days). The rats were sacrificed and everted sacks of the jejunum were made and incubated in EGF solution. Western blot analysis of mucosal homogenates showed that the amount of phosphotyrosyl EGF receptor in the starved and MTX-treated groups was, respectively, about 1.5 times and 2 times that in the control group. The mucosal permeability in the starved and MTX treated groups also increased and varied directly with the amount of phosphotyrosyl EGF receptor. These results suggest that in the adult rat intestine, luminal EGF may play a role only under tissue damage, where enhanced permeability permits the EGF to pass through the mucosa and bind to its receptor on the basolateral membrane.

Enhancement of aquaporin-3 by vasoactive intestinal polypeptide in a human colonic epithelial cell line.

BACKGROUND: Vasoactive intestinal polypeptide (VIP) plays an important role in water transport in the intestine. Several specialized channels termed aquaporins (AQP) facilitate water transport in the gastrointestinal tract. Aquaporin-3 localizes to epithelial cells in the human small intestine and colon. However, the regulatory mechanisms underlying the functions of AQP3 remain unclear. To characterize the regulation of AQP3 expression by VIP, we studied messenger (m)RNA expression, protein expression and DNA binding activity in a human colonic epithelial cell line, HT-29. METHOD: Human colonic epithelial cells, HT-29, were incubated with VIP (10-12-10-7 M). The cells were treated with protein kinase-A (PK-A) inhibitors (H-89, H-9) or chloride channel-blockers (diphenylamine-2-carboxylate (DPC), 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPD)). The expression of AQP3 mRNA and protein was determined by Northern blot and Western blot, respectively. The DNA-binding activities of cyclic adenosine monophosphate (cAMP) response elements/activating transcription factor (CRE/ATF)) in the nuclear extract were determined by electrophoretic mobility shift assay. RESULTS: Aquaporin-3 mRNA was up-regulated at a concentration of 10-10 M VIP. The expression of AQP3 protein was enhanced at 3 h after addition of VIP. The PK-A inhibitors (H-89, H-9) inhibited the expression of AQP3 mRNA enhanced by VIP and cAMP. The gel shift assay of CRE/ATF in HT-29 cells revealed a single band. CONCLUSION: These results indicate that VIP upregulated the expression of AQP3 mRNA and protein, and that a cAMP-dependent pathway mediated this effect in a human colonic epithelial cell line, HT-29.

Detection of enriched Thomsen-Friedenrich antigen on IgA1 from IgA nephropathy patients.

BACKGROUND: Human serum IgA1 has a mucin-like structure on its hinge portion which is composed of a mucin-type sugar chain and amino acid sequence rich in proline, serine and threonine. There are incompletely glycosylated O-linked oligosaccharide(s) on the IgA1 hinge region in some nephropathy patients. METHODS: We made a detailed analysis of the incompleteness of the sugar chain by digesting IgA1 with various glycosidases. To verify the incompleteness of the sugar chains, the galactosamine/glucosamine ratio (O/N ratio) was introduced as a specific value for each IgA1 preparation. RESULTS: When IgA1 from serum was treated with alpha-N-acetylgalactosaminidase and/or neuraminidase or endo-beta-N-acetylglucosaminidase H (Endo-H), the O/N ratio did not change. However, endo-alpha-N-acetylgalactosaminidase (Endo-A) reduced the O/N ratio of IgA1 from the IgA nephropathy patient whereas before treatment, the O/N ratio had been similar in the normal control and IgA nephropathy. CONCLUSIONS: This result means there is a small amount of the unsubstituted and the sialylated N-acetylgalactosamine residues (Tn and sialyl Tn antigen, respectively), and abundant asialo Galbeta1,3GalNAc (TF antigen) in the IgA1 molecule. In view of the incompleteness of the IgA1 sugar chain, the decrease in the sialic acid content of the mucin-type sugar chain on IgA1 from an IgA nephropathy patient became obvious in this experiment.

Alteration in expression of polyamine and glucose-related enzyme mRNA after small bowel resection in the rat residual ileum.

The adaptive hyperplasia of the residual intestine after a massive bowel resection is not fully understood. We investigated the alterations in polyamine and glucose-related enzyme mRNA expression during intestinal adaptation. Six-week-old male Wistar rats underwent an 80% resection of the small intestine. The residual ileum was removed on the preoperative day (control) and on postoperative day (POD) 1, 3, 5 and 7. The total RNA was extracted from the mucosa, and a Northern blot analysis was performed. In the residual small intestine, the expression of polyamine synthesis enzymes, ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) mRNAs were increased on POD 1. The expression of polyamine degradation enzymes diamine oxidase (DAO) and spermidine/spermine N1-acetyltransferase (SSAT) mRNA did not change dramatically. Antizyme-1 (AZ-1) mRNA was significantly increased on POD 1. The mRNA expression of glucose absorption and metabolism-related proteins, including the Na+-dependent D-glucose cotransporter (SGLT1), fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase (Fru-6-P,2-kinase/Fru-2,6-Pase) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were only slightly changed on POD 1. The enzymes responsible for polyamine biosynthesis but not catabolism were upregulated at the translational level in enterocytes after a small bowel resection. The expression of glucose transport and glycolysis enzyme mRNAs did not increase after a small bowel resection.

In vitro alterations in fecal short chain fatty acids and organic anions induced by the destruction of intestinal microflora under hypotonic and aerobic conditions.

The pathogenesis of inflammatory bowel disease (IBD) remains unknown. It has been suggested that luminal factors such as the microflora, short chain fatty acids (SCFAs), food antigens and so on play important roles in the disease progression. Many reports have revealed alterations in the SCFA and organic acid concentration of the colon, especially increased lactate and decreased butyrate, in IBD patients. The mechanisms responsible for these alterations, however, remain unclear. Therefore, the effects of aerobic conditions on the alterations in the SCFA and organic anion levels in the feces was evaluated. Fecal specimens were collected from 5 healthy volunteers. Under aerobic condition, a mixture of feces and distilled water was incubated in 37 degrees C for 1 and 3 h. The pH, osmotic pressures, the concentrations of potassium, bicarbonate, SCFA and organic anion, and the activities of alpha-amylase and lactate dehydrogenase (LDH) in the mixture were then measured. We also examined any changes in the microscopic microflora under the hypotonic and aerobic conditions. The results showed the osmotic pressure, and the concentrations of lactate and SCFAs (formate, acetate, propionate and n-valerate) were progressively increased with longer incubation times, and reached a statistically significant difference. In particular, the ratios of lactate, succinate and n-valerate after 1 and 3 h of incubation increased remarkably. In contrast, the electrolyte levels and both alpha-amylase and LDH activities were not altered significantly. Microscopically, the microflora in the mixture decreased with prolonged incubation times. These data suggest that under these in vitro conditions, the organic anion and SCFA levels in the feces easily increased. It is probable that the alterations in the SCFA and organic anion levels in IBD patients may be partly due to intracellular components derived from microflora destroyed under hypotonic and aerobic conditions in the colonic lumen, for example caused by mucosal bleeding. These alterations may influence the pathogenesis and progression of IBD.