Stool and urine trefoil factor 3 levels: associations with symptoms, intestinal permeability, and microbial diversity in irritable bowel syndrome.

Previously we showed that urine trefoil factor 3 (TFF3) levels were higher in females with irritable bowel syndrome (IBS) compared to non-IBS females. To assess if TFF3 is associated with symptoms and/or reflect alterations in gastrointestinal permeability and gut microbiota in an IBS population, we correlated stool and urine TFF3 levels with IBS symptoms, intestinal permeability, stool microbial diversity and relative abundance of predominant bacterial families and genera. We also tested the relationship of stool TFF3 to urine TFF3, and compared results based on hormone contraception use. Samples were obtained from 93 females meeting Rome III IBS criteria and completing 4-week symptom diaries. TFF3 levels were measured by ELISA. Permeability was assessed with the urine lactulose/mannitol (L/M) ratio. Stool microbiota was assessed using 16S rRNA. Stool TFF3, but not urine TFF3, was associated positively with diarrhoea and loose stool consistency. Higher stool TFF3 was also associated with lower L/M ratio and microbial diversity. Of the 20 most abundant bacterial families Mogibacteriaceae and Christensenellaceae were inversely related to stool TFF3, with only Christensenellaceae remaining significant after multiple comparison adjustment. There were no significant relationships between stool or urine TFF3 levels and other symptoms, nor between stool and urine levels. In premenopausal females, urine TFF3 levels were higher in those reporting hormone contraception. Collectively these results suggest that higher stool TFF3 levels are associated with IBS symptoms (loose/diarrhoeal stools), lower gut permeability, and altered stool bacteria composition (decreased diversity and decreased Christensenellaceae), which further suggests that TFF3 may be an important marker of host-bacteria interaction.

Relationships of abdominal pain, reports to visceral and temperature pain sensitivity, conditioned pain modulation, and heart rate variability in irritable bowel syndrome.

BACKGROUND: Irritable bowel syndrome (IBS) is a heterogeneous condition with a number of pathophysiological mechanisms that appear to contribute to symptom chronicity. One of these is altered pain sensitivity. METHODS: Women between ages 18-45 were recruited the community. Of those enrolled, 56 had IBS and 36 were healthy control (HC) women. Participants completed questionnaires, kept a 4-week symptom diary and had a 12-h Holter placed to assess nighttime heart rate variability including high frequency power (HF), low frequency power (LF), and total power (TP). At mid-follicular phase approximately 80% of women completed a thermal pain sensitivity test with conditioned pain modulation and visceral pain sensitivity using a water load symptom provocation (WLSP) test. KEY RESULTS: As expected, daily abdominal pain was significantly higher in the IBS compared to HC group. There were no differences between the bowel pattern subgroups (IBS-diarrhea [IBS-D], IBS-constipation plus mixed [IBS-CM]). Thermal pain sensitivity did not differ between the IBS and the HC groups, but was significantly higher in the IBS-CM group than the IBS-D group. In the WLSP test, the IBS group experienced significantly more symptom distress than HCs and the IBS-CM group was higher than the IBS-D group. Heart rate variability indicators did not differ between the groups or IBS subgroups. Daily abdominal pain was positively correlated with LF and TP in the IBS group. CONCLUSIONS & INFERENCES: Despite similar levels of abdominal pain in IBS, the IBS-CM group demonstrated greater sensitivity to both thermal and visceral testing procedures.

Numerical Modeling of Mechanical Behavior for Buried Steel Pipelines Crossing Subsidence Strata.

This paper addresses the mechanical behavior of buried steel pipeline crossing subsidence strata. The investigation is based on numerical simulation of the nonlinear response of the pipeline-soil system through finite element method, considering large strain and displacement, inelastic material behavior of buried pipeline and the surrounding soil, as well as contact and friction on the pipeline-soil interface. Effects of key parameters on the mechanical behavior of buried pipeline were investigated, such as strata subsidence, diameter-thickness ratio, buried depth, internal pressure, friction coefficient and soil properties. The results show that the maximum strain appears on the outer transition subsidence section of the pipeline, and its cross section is concave shaped. With the increasing of strata subsidence and diameter-thickness ratio, the out of roundness, longitudinal strain and equivalent plastic strain increase gradually. With the buried depth increasing, the deflection, out of roundness and strain of the pipeline decrease. Internal pressure and friction coefficient have little effect on the deflection of buried pipeline. Out of roundness is reduced and the strain is increased gradually with the increasing of internal pressure. The physical properties of soil have a great influence on the mechanical properties of buried pipeline. The results from the present study can be used for the development of optimization design and preventive maintenance for buried steel pipelines.

CT colonography in a Korean population with a high residue diet: comparison between wet and dry preparations.

AIM: To compare wet and dry preparation methods for computed tomography colonography (CTC) in terms of preparation quality, interpretation time, and diagnostic performance for polyp detection in a population with a high residue diet. MATERIALS AND METHODS: Eighty-six patients were divided into two groups. Group 1 (n=24) received a wet preparation of 4l polyethylene glycol (PEG) solution, and group 2 (n=62) received a dry preparation of phosphor-soda. Abnormal findings, including polyps, and the time required to interpret the CTC images in both groups were documented by a radiologist. CTC findings were compared to those of colonoscopy as a reference standard. Two radiologists evaluated the quality of CTC with regard to residual fluid, faeces, and colonic distension using a four-point scale in consensus. Statistical differences for residual fluid, faeces, distensibility on CTC, and interpretation time between the two groups were analysed. The diagnostic performance of CTC in both groups was also compared. RESULTS: One-hundred and ninety polyps in 70 patients were identified using colonoscopy. Regarding the quality of images produced the wet preparation was significantly better than the dry preparation (p<0.05). The average interpretation time was significantly shorter for the wet group (11.7 min) than the dry group (16.4 min) (p<0.05). For per-patient analysis, the positive predictive value (PPV) was significantly better for the wet (100%) than the dry group (79.6%; p=0.025). Sensitivities and PPV for >or=10 mm polyps were comparable between two groups (p>0.05). CONCLUSION: In a population with a high-residue diet, CTC with wet preparation can be interpreted in a time-efficient manner and is comparable with CTC with dry preparation.

Trace but not delay fear conditioning requires attention and the anterior cingulate cortex.

Higher cognitive functions such as attention have been difficult to model in genetically tractable organisms. In humans, attention-distracting stimuli interfere with trace but not delay conditioning, two forms of associative learning. Attention has also been correlated with activation of anterior cingulate cortex (ACC), but its functional significance is unclear. Here we show that a visual distractor interferes selectively with trace but not delay auditory fear conditioning in mice. Trace conditioning is associated with increased neuronal activity in ACC, as assayed by relative levels of c-fos expression, and is selectively impaired by lesions of this structure. The effects of the ACC lesions are unlikely to be caused by indirect impairment of the hippocampus, which is required for mnemonic aspects of trace conditioning. These data suggest that trace conditioning may be useful for studying neural substrates of attention in mice, and implicate the ACC as one such substrate.

Reciprocal expressions of cyclin E and cyclin D1 in hepatocellular carcinoma.

Deregulation of the cell cycle by overexpression of G1 cyclins, cyclin E and cyclin D1 genes, has been demonstrated to be a prerequisite for the development of human cancer. Recently, cyclin E is proposed to be sufficient for the progression of the G1 cell cycle without cyclin D1. Here we show that the proposed model system was specifically present in human hepatocellular carcinoma (HCC) unlike other human cancers. Of 31 HCC tissues analyzed, 21 (67.7%) exhibited an overexpression of cyclin E protein. In contrast to cyclin E gene expression, cyclin D1 expression was strongly downregulated in 19 (61.2%) HCCs. Interestingly, 65% of HCC tissues with overexpression of the cyclin E gene exhibited downregulation of cyclin D1, suggesting reciprocal deregulation of these cyclins in the G1 progression of the cell cycle. Southern blot analysis proved the amplification of cyclin E gene in HCC with a high level of overexpression. The present findings suggest that the reciprocal deregulation of cyclin E lacking cyclin D1 expression might play a role in G1 progression and the development of HCC.

Cannabinoid modulation of time estimation in the rat.

Cannabinoids have been implicated in a variety of cognitive processes in humans, including attention, learning, memory, and time estimation. However, studies of the effects of cannabinoids on rodent behavior have focused on motor, learning, and memory tasks. To assess cannabinoid effects on time perception, this study examined whether systemically administered cannabinoid receptor agonists and a cannabinoid receptor antagonist influenced rats' performance of a time interval estimation task based on a fixed-interval schedule (a "peak procedure"). Both cannabinoid agonists WIN 55,212-2 and delta9-tetrahydrocannabinol shortened the modal response time, and cannabinoid antagonist SR 141716A lengthened the modal response time. Secondary measures of the shape of the response distribution were not influenced by any of the drugs, suggesting that the response distribution shifts were not artifacts of drug side effects. Therefore, these experiments argue for the involvement of endogenous cannabinoids in time estimation.

A performance-dependent adjustment of the retention interval in a delayed non-matching-to-position paradigm differentiates effects of amnestic drugs in rats.

Operant delayed non-matching-to-position (delayed non-matching-to-position) tasks have been widely used as tests of working memory in rats, but have suffered some loss in sensitivity to differentiating selective mnemonic from non-mnemonic deficits due to floor and ceiling effects. To circumvent this problem, a novel delayed non-matching-to-position was developed in which the retention interval was adjusted on a trial-by-trial basis to hold performance accuracy at an intermediate value. The present study assessed the effects of three amnestic drugs in this delayed non-matching-to-position. Rats were administered (i.p.) NMDA receptor antagonist ((5R,10S)-(+)-5-Methyl-10, 11-dihydro-5H-dibenzo[a,d,] cyclohepten-5,10-imine (Dizocilpine or MK-801), muscarinic receptor antagonist (-)-scopolamine hydrobromide (scopolamine), or cannabinoid receptor agonist ((R)-(+)-[2, 3-Dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1, 4-benzoxazin-6-yl]-1-naphthalenylmethanone) (WIN 55, 212-2). At high doses, both MK-801 (0.12-0.25 mg/kg) and scopolamine (0.25 mg/kg) produced deficits not selective to working memory. At low doses, scopolamine (0.06-0.12 mg/kg) and MK-801 (0.06 mg/kg) produced no deficits in any mnemonic or secondary measures. WIN 55, 212-2 produced deficits at 2.0 mg/kg that were consistent with a specific impairment of working memory. Using this particular delayed non-matching-to-position revealed that consistent changes in performance accuracy at the short retention interval were evident for scopolamine and MK-801, at times in the absence of changes in response tendency, which are consistent with an interpretation that these drugs produce general deficits in reference or procedural memory. In contrast, cannabinoid-induced deficits in choice accuracy support previous reports of delay-dependent deficits. Together, these data suggest that this delayed non-matching-to-position task is able to differentiate deficit patterns of amnestic drugs, and isolate the effects of motivational side effects of drugs from working memory measurement.

Specificity of memory measures in an adjusting delayed nonmatching-to-position task for rats.

While the delayed nonmatching-to-position (DNMTP) behavioral paradigm has often been used by neuroscientists to assess working memory in rats, its measure of working memory is compromised by floor and ceiling effects. Specifically, these floor and ceiling effects undermine the ability to detect a significant interaction in a two-factor repeated-measures ANOVA, which is required in order to conclude that impaired performance has resulted from disrupted working memory and not from a change in another psychological process (i.e. that the impairment is 'specific' to working memory). The present study was conducted to evaluate if these limitations could be overcome in a DNMTP by adjusting the length of time that the rat was required to remember (the 'delay') so as to avoid the floor and ceiling. The general procedure for two experiments presented subjects with trials where there was either a minimal (1 s) delay or a longer delay of varying length, with the goal of maintaining nonmatch-to-position accuracy near 75%. The procedure was such, that if the average accuracy was at or above 75%, then the next trial would be a long-delay trial. If it were below 75%, then the next trial would be a 1 s delay trial. In the first experiment, the subjects were presented with trials where the value of the longer delay was systematically varied between-sessions. This was done to simulate the faster rate of forgetting found in persons with amnesia. DNMTP accuracy diminished at the longer delay but not the short delay as the second interval was lengthened. However, other measures, including accuracy at the 1 s delay, discrimination accuracy during the sample phase, and the number of trials completed per session, did not change. This experiment suggested that this DNMTP could precisely measure small changes in the rate of forgetting. In a second experiment, a potential non-mnemonic confound, the level of motivation, was directly manipulated by giving the water-restricted subjects access to water immediately prior to the start of the session. The number of trials completed per session diminished, but the accuracy at both the 1 s and the longer delays did not change. These results indicated that the measures of working memory in this DNMTP were insensitive to changes in motivation. Together, these experiments confirmed that adjusting the delay in the DNMTP improves the validity of the measures in this task.

Clinical usefulness of serum and plasma vascular endothelial growth factor in cancer patients: which is the optimal specimen?

Vascular endothelial growth factor (VEGF) is secreted by various human cancer cells and plays a key role in cancer angiogenesis and metastasis. Recently, evidence of VEGF storage in blood cells including platelets has been reported. The serum VEGF levels were reported to increase during clotting as a result of its release from platelets, and plasma sample instead of serum was recommended for measuring the circulating VEGF more accurately. However, platelets have been implicated in tumor metastasis since circulating tumor cells forming aggregates with platelets were observed. The purpose of this study was to clarify which is an optimal specimen to measure VEGF in cancer patients, serum or plasma. We measured serum and plasma VEGF levels and platelet counts in 173 cancer patients and 42 healthy people, and found that serum VEGF levels were significantly higher than matched plasma VEGF and the VEGF difference (serum VEGF - plasma VEGF) correlated with platelet counts (r=0.624, p<0.05) in both cancer patients and healthy controls. We selected cancer patients with normal platelet counts (130-400x103/microl, Plt-normal cancer group). Interestingly, serum VEGF levels were higher in Plt-normal cancer group than in healthy controls. The theoretical platelet-derived VEGF in serum, calculated based on actual blood platelet counts (pg per 106 platelets), was also significantly higher in Plt-normal cancer group than in normal controls. It is, therefore, suggested that, although the serum VEGF levels are affected by blood platelets, platelet-derived VEGF also reflect biology of cancer cells, and that serum would be the more useful specimen for measurement of circulating VEGF in cancer patients for prognosis.

Attenuation of cardiac but not vascular component in baroreflex of spontaneously hypertensive rats.

The cardiac and vascular components of the baroreceptor reflex in spontaneously hypertensive rat (SHR) and stroke-prone spontaneously hypertensive rat (SHRSP) were compared against their counterparts in normotensive Wistar Kyoto rat (WKY). SHR, SHRSP and WKY of 12-16 weeks old were chronically instrumented for intra-arterial recording of blood pressure. Intravenous injections of phenylephrine and nitroprusside were used to challenge their baroreflex. The products of blood pressure change and the half time required for the pressure to return to the control value were used as the quantitative estimation of the blood pressure stabilizing capability. The cardiac component of the baroreflex was obtained from the change in the blood pressure stabilizing capability after blockade of beta and muscarinic receptors by atenolol and atropine, respectively. The vascular component was obtained by subtracting the cardiac component from the total stabilizing capability which was the difference after blockade with a ganglionic transmission blocker, hexamethonium. We found the cardiac component of the baroreflex of the hypertensive rats was significantly less sensitive than that of the WKY. In contrast, the vascular component of the baroreflex of the three strains did not differ significantly. Therefore, we concluded that the 12-16 week old SHRs were able to maintain a stable blood pressure due to the intact vascular component of the baroreflex.

Continuous culture as a tool for investigating the growth physiology of heterotrophic hyperthermophiles and extreme thermoacidophiles.

Although there is great scientific and technological interest in examining the physiology and bioenergetics of microorganisms from extreme environments, difficulties encountered in their cultivation and lack of genetic systems hampers the investigation of these issues. As such, we have adapted methods for continuous cultivation of mesophilic organisms to extremes of temperature and pH to study extremophiles. Since the risk for contamination of extremophilic continuous cultures is relatively small, long-term, steady state experiments investigating physiological response to culture perturbations are possible. Experiments along these lines have provided insights into the significance of specific enzymes in the metabolism of particular substrates, in addition to providing a better understanding of stress response and unusual physiological characteristics of hyperthermophilic and extremely thermoacidophilic microorganisms. Several examples are provided here, including the thermal stress response of Metallosphaera sedula (T(opt) 74 °C) growing at pH 2.0, and the response of the heterotrophic hyperthermophiles Pyrococcus furiosus (T(opt) 98 °C), Thermococcus litoralis (T(opt) 88 °C) and T. maritima (T(opt) 80 °C) to changes in growth medium. Also discussed will be how the same experimental systems have been used to study exopolysaccharide production and biofilm formation by hyperthermophilic heterotrophs and facilitated the estimation of bioenergetic parameters for these organisms under a variety of growth conditions. Continuous culture, used in conjunction with genome sequence information, two-dimensional gel electrophoresis and differential gene expression, can provide important insights into the metabolism of high temperature extremophiles.

Acquired Thermotolerance and Stressed-Phase Growth of the Extremely Thermoacidophilic Archaeon Metallosphaera sedula in Continuous Culture.

The response of an extremely thermoacidophilic archaeon, Metallosphaera sedula (growth temperature range, 50 to 79(deg)C; optimum temperature, 74(deg)C; optimum pH, 2.0), to thermal stress was investigated by using a 10-liter continuous cultivation system. M. sedula, growing at 74(deg)C, pH 2.0, and a dilution rate of 0.04 hr(sup-1), was subjected to both abrupt and gradual temperature shifts in continuous culture to determine the responses of cell density levels and protein synthesis patterns. An abrupt temperature shift from 74 to 79(deg)C resulted in little, if any, changes in cell density and a small increase in total protein per cell. When the culture temperature was shifted further to 80.5(deg)C, cell density dropped to below 5 x 10(sup6) cells/ml from 10(sup8) cells/ml, leading to washout of the culture. Operation at this temperature and slightly higher temperatures, however, could be achieved by exposing the culture to thermal stress more gradually (0.5(deg)C increments). As a result, stable operation could be maintained at temperatures of up to 81(deg)C, and the washout temperature could be increased to 82.5(deg)C. Continuous culture operation at 81(deg)C for 100 h (stressed phase) led to an approximately sevenfold lower steady-state cell density than that observed for operation at or below 79(deg)C. However, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis (both one and two dimensional) revealed significantly higher levels (sixfold increase) of a 66-kDa stress response protein (MseHSP60), immunologically related to Thermophilic Factor 55 from Sulfolobus shibatae (J. D. Trent, J. Osipiuk, and T. Pinkau, J. Bacteriol. 172:1478-1484, 1990). If the acclimated culture was returned to a lower temperature (i.e., 74(deg)C), the amount of MseHSP60 returned to levels observed prior to thermal acclimation. Furthermore, when the previously acclimated culture (at 81(deg)C) was shifted back from 74 to 81(deg)C, without going through gradual acclimation steps, the result was the immediate onset of washout, suggesting no residual thermotolerance. This study shows that gradual thermal acclimation of M. sedula could only extend the temperature range of stable growth for this organism by 2(deg)C above its maximal growth temperature, albeit at reduced cell densities. Also, this investigation illustrates the utility of continuous culture for characterizing heat shock response and assessing maximum growth temperatures for extremely thermophilic microorganisms.

Hepatitis C virus genotypes in Korea and their relationship to clinical outcome in type C chronic liver diseases.

OBJECTIVES: The relationship between HCV genotype and the development of more serious liver disease has not been clearly established. This study was to investigate the distribution pattern of HCV genotypes in Korea and their relationship to the viremic level and to progression of chronic liver disease. METHODS: Study population was 217 patients with type C chronic liver disease. They were divided into 4 groups; 83 patients with near-normal ALT (group 1), 64 patients with elevated ALT (group 2), 20 patients with decompensated liver cirrhosis (group 3) and 50 patients with hepatocellular carcinoma (group 4). HCV genotypes were determined by reverse transcription polymerase chain reaction (RT-PCR) using mixed primer sets, and then the fidelity of genotyping was confirmed by cloning and sequencing. HCV RNA concentration was measured by quantitative competitive RT-PCR for 23 patients in group 2. RESULTS: The genotypes could be determined in 166 (76%) out of 217 patients. Type 1b and type 2a were predominantly occurring over the other types in somewhat similar frequency (45% and 51%, respectively). The genotype distribution of type 1b and 2a among four different groups showed 42% and 54% in group 1, 49% and 45% in group 2, 53% and 47% in group 3 and 41% and 57% in group 4; thus there was no significant difference in genotype distribution among 4 different disease groups. However, the viremia levels in patients with genotype 1b infection were significantly higher than those with genotype 2a. CONCLUSION: Genotype 2a infection is as prevalent as genotype 1b in Korea, and genotype 2a infection may pose no less risk for progression of disease despite lower replication level than genotype lb infection.

Recovery of proteins and amino acids from reverse micelles by dehydration with molecular sieves.

A new method is presented to precipitate proteins and amino acids from reverse micelles by dehydrating the micelles with molecular sieves. Nearly complete precipitation is demonstrated for alpha-chymotrypsin, cytochromec, and trytophan from 2-ethylhexyl sodium sulfosuccinate (AOT)/isooctane/water reverse micelle solutions. The products precipitate as a solid powder, which is relatively free of surfactant. The method does not require any manipulation of pH, ionic strength, temperature, pressure, or solvent composition, and is applicable over a broad range of these properties. This general approach is compared with other techniques. This general approach is compared with other techniques for the recovery of biomolecules from reverse micelles. (c) 1994 John Wiley & Sons, Inc.

Predominant etiologic association of hepatitis C virus with hepatocellular carcinoma compared with hepatitis B virus in elderly patients in a hepatitis B-endemic area.

BACKGROUND: It has been thought that the mean age of patients with hepatitis B surface antigen (HBsAg) positivity and hepatocellular carcinoma (HCC) is younger than that of patients who test positive for anti-hepatitis C virus. This study was to assess the relative etiologic association of hepatitis B virus (HBV) and hepatitis C virus (HCV) with HCC, according to age-specific groups. METHODS: A total of 336 Korean patients with HCC were enrolled in the study; the mean age was 53.7 +/- 9.1 years, and the male-to-female ratio was 5.9. HBV serologic markers and anti-HCV were tested using commercially available radioimmunoassay and enzyme immunoassay kits, respectively. RESULTS: Two hundred thirty-three (69.3%) patients tested positive for HBsAg; anti-HCV was positive in 42.7% of 103 patients with HBsAg-negative disease and in 17.0% of all patients with HCC. Coinfection rate of HBV and HCV in the total number of patients with HCC was 3.9%. The mean age was 51.6 years for patients with HBsAg-positive disease and 60.4 years for patients with anti-HCV-positive disease. The ratio of HBsAg positivity compared with anti-HCV positivity in patients with HCC was 29.7 for patients younger than 50 years; 0.9 for patients older than 61 years; and 5.2 for patients in their 50s. CONCLUSIONS: The authors conclude that although HBV infection plays a more important etiologic role in the development of HCC in Korea than does HCV, HCV is no less frequently involved than HBV in the etiology of HCC for patients older than 61 years. Thus, in surveillance programs for the early detection of HCC, closer attention should be directed to the patients with anti-HCV-positivity and chronic liver disease, especially patients older than 61 years, even in a hepatitis-B-endemic area.

Polymorphic reticulosis with colonic lesion--a case report.

A 38-year-old man was admitted with a high fever, sore throat, and right upper quadrant pain. Nine months before his admission, he had undergone right hemicolectomy under the impression of intestinal lymphoma. But there had been no evidence of lymphoma on microscopic examination. Under the postoperative diagnosis of inflammatory bowel disease, corticosteroid therapy was tried without response. On the follow-up colonoscopic examination, an ovoid ulcer, with convergence of the surrounding mucosal folds at the descending colon and an irregularly shaped ulcer at the ileocolic anastomotic site, were found. The colonoscopic diagnosis was Behcet's colitis. After pathologic slides of biopsy and surgical specimens obtained from the palatine tonsil and colon were reviewed, the diagnosis of polymorphic reticulosis was made. The patient received anticancer chemotherapy, including cyclophophamide and glucocorticosteroid. To date, colonic involvement of polymorphic reticulosis has not been reported. Because of the similarity of the colonoscopic findings to those of Behcet's colitis, polymorphic reticulosis should be included in the differential diagnosis of inflammatory bowel disease. We assume that this is the first case of polymorphic reticulosis involving the colon with characteristic colonoscopic findings.