Effectiveness of an 'half elemental diet' as maintenance therapy for Crohn's disease: A randomized-controlled trial.

BACKGROUND: Although thiopurines have a proven role in maintenance therapy for Crohn's disease, an alternative therapy is needed for patients intolerant or resistant to thiopurines. AIM: To evaluate the effectiveness of home enteral nutrition as a maintenance therapy regimen in which half of the daily calorie requirement is provided by an elemental diet and the remaining half by a free diet. We refer to this home enteral nutrition therapy as 'half elemental diet'. METHODS: Between 2002 and 2005, 51 patients in remission from two hospitals were randomly assigned to a half elemental diet group (n = 26) or a free diet group (n = 25). The primary outcome measure of this study was the occurrence of relapse over the 2-year period. RESULTS: The relapse rate in the half elemental diet group was significantly lower [34.6% vs. 64.0%; multivariate hazard ratio 0.40 (95% CI: 0.16-0.98)] than that in the free diet group after a mean follow-up of 11.9 months. Compliance was similar in the two groups. No adverse event occurred in any of the patients throughout the study. CONCLUSION: This randomized-controlled trial shows the effectiveness of an half elemental diet, which is a promising maintenance therapy for Crohn's disease patients.

In vitro cytotoxicity of a novel antitumor antibiotic, spicamycin derivative, in human lung cancer cell lines.

Spicamycin (SPM), produced by Streptomyces alanosinicus, induces potent differentiation in a human leukemia cell line, HL60. One of the derivatives of SPM (SPM-D), KRN5500, has a wide range of antitumor activity against human cancer cell lines. We examined the cytotoxicity of SPM-D in small and non-small cell lung cancer cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and colony assays. SPM-D was active against a wide range of lung cancer cell lines. All three cisplatin (CDDP)-resistant cell lines established in our laboratory (PC-9/CDDP, PC-14/CDDP, and H69/CDDP) showed collateral sensitivity to SPM-D with relative resistance values of 0.43, 0.34, and 0.32, respectively. Intracellular SPM-D in PC-14/CDDP was 35% higher than that for PC-14 suggesting that intracellular accumulation can explain the collateral sensitivity to SPM-D at least in PC-14/CDDP. On the other hand, in PC-9/CDDP cells, no increase of intracellular SPM-D accumulation was observed, but the conversion ratio of a metabolite (the amino nucleoside moiety of spicamycin binding with glycine, SAN-G) from SPM-D evaluated by TLC was higher as compared with that of parental PC-9 cells (45.5% versus 37%; PC-9/CDDP versus PC-9). The increased intracellular metabolism of SPM-D could explain the mechanism of collateral sensitivity in PC-9/CDDP cisplatin-resistant cell lines. To elucidate the determinant of the SPM-D-induced cytotoxicity, we established SPM-D-resistant cell lines, PC-9/SPM-D, PC-14/SPM-D, and H69/SPM-D, by exposing cells to stepwise increases in SPM-D concentration. The relative resistances of these sublines were more than 5000, 46.6, and 37.8 times those of the parental cell lines, respectively. The intracellular concentration of the active metabolite, SAN-G, was found to be decreased in the SPM-D-resistant sublines. This result indicates that the intracellular metabolism of SPM-D to SAN-G is one of the determinants of cellular sensitivity to SPM-D in these SPM-D-resistant cell lines. In conclusion, both drug accumulation and metabolism may contribute to the sensitivity/resistance to SPM-D and both may merit investigation.

Cell turnover in normal cycling human ovary.

We investigated cell proliferation and apoptosis in 37 normal cycling human ovaries to determine cell turnover in the various stages of follicular and luteal development. We examined cell proliferation by immunostaining for both Ki67 and proliferative cell nuclear antigen (PCNA) and by silver staining of nucleolar organizer regions (AgNORs). Apoptosis was examined by 3'-hydroxy nick-end labeling and by immunostaining of an apoptosis-related antigen, Ley. The labeling indexes of Ki67, PCNA, and AgNORs were significantly increased in antral follicles. However, there were no significant differences in the labeling index of Ki67, PCNA, and AgNORs between dominant and nondominant follicles, including nonovulated follicles in the luteal phase. These results indicate that the transformation of granulosa cells from quiescence to active growth is important in early folliculogenesis. Immunoreactivity for Ki67 and PCNA were observed predominantly in the functioning corpus luteum, but not in the degenerating corpus luteum, indicating proliferation only during the luteal phase. Immunoreactivity for Ley and nick end-labeling reactive cells were not observed in the follicular and luteal phases, except for scattered cells in the degenerating corpus luteum. This may be because of the relatively long process of human follicular growth and atresia.

Improvement by eicosanoids in cancer cachexia induced by LLC-IL6 transplantation.

Cachexia frequently occurs in the late stages of cancer, and is difficult to manage. We previously reported that interleukin-6 (IL-6) cDNA transfection into Lewis lung carcinoma (LLC-IL6) induced cachexia-like symptoms in C57BL/6 mice. This was thought to be a useful experimental model of cancer cachexia. We have examined the effects of two eicosanoids, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), in order to evaluate whether they could relieve cachexia. LLC-IL6-bearing animals were divided into three treatment groups receiving DHA, EPA or water as the control; 80-microliter samples of these compounds (purity > 95%) were administered orally by catheter daily starting 7 days after tumor transplantation. Tumor growth curves were similar in the three groups. There were no differences in water or food intake in the three groups. However, body weight, a marker of cachexia, was significantly higher in treated mice than in the control group. Sixteen days after tumor transplantation, the mean body weight was 17.45 g (P < 0.05), 17.2 g and 16.41 g in the groups receiving DHA, EPA and water respectively. The eicosanoids did not affect serum levels of IL-6. Ubiquitination of muscle protein, a marker of proteolysis coupled to cachexia, was compared in LLC-IL6- and LLC-transplanted mice. The eicosanoids prevented the ubiquitination of approximately 180 kDa protein. These results suggest that eicosanoids may prevent the cachexia mediated by IL-6.

In vitro and in vivo growth of B16F10 melanoma cells transfected with interleukin-4 cDNA and gene therapy with the transfectant.

In an attempt to develop the most effective cytokine gene therapy, we transfected mouse interleukin(IL)-2, mouse IL-4, and human IL-6 cDNAs into mouse melanoma cells, B16F10. Transfection with IL-4 cDNA decreased the tumorigenicity of B16F10 most strongly. We investigated whether gene therapy with IL-4-transfected B16F10 cells was possible. Flow-cytometric analysis showed that major histocompatibility complex class I and II expression in B16F10 and IL-4-cDNA-transfected B16F10 (B16F10-IL4) cells did not differ. Doubling times of B16F10 and B16F10-IL4 were 20.1 and 21.1 h respectively. The growth of B16F10 cells was retarded if C57BL/6 mice were inoculated with B16F10-IL4 at the contralateral sides. When 5 x 10(5) B16F10 cells were transplanted subcutaneously into the flanks of C57BL/6 mice, they all developed a tumor mass, whereas no tumor masses formed in those transplanted with B16F10-IL4 cells within 60 days. No nude, severe combined immunodeficient or beige mice were able to reject parental B16F10 or B16F10-IL4 cells, although, B16F10-IL4 tumor growth in all these immunodeficient mice was slower than that of B16F10. Therefore, we concluded that T and natural killer cells are necessary for rejection of B16F10-IL4 tumor cells.

Plasma glicentin in diabetic and gastrectomized patients.

Recent successful synthesis of human glicentin prompted us to establish an immunoassay method for determination of human glicentin in plasma. Human glicentin in plasma was measured using a newly developed sandwich ELISA. The mean fasting levels of human glicentin were 18.6+/-2.4 and 19.7+/-2.1 pM in normal subjects and diabetic patients, respectively. In diabetic patients with renal failure, plasma glicentin was elevated, exceeding 100 pM. In normal subjects, plasma glicentin increased to a peak level of about 130 pM at 60 min after an oral glucose load, and then decreased. In patients who underwent gastrectomy, plasma glicentin rapidly increased to a peak of about 300 pM at 30 min after oral glucose load. In a patient with short bowel syndrome plasma glicentin did not change following an oral glucose load. These results correspond with previous findings for gut glucagon-like immunoreactive materials (GLI) or enteroglucagon. We conclude that glicentin is secreted from the small intestine in response to intraluminal glucose stimulation in humans.

In vivo screening models of cisplatin-resistant human lung cancer cell lines using SCID mice.

In vivo screening models of a cisplatin (CDDP)-resistant human small-cell lung cancer cell (SCLC) line, H69/CDDP, and a non-small-cell lung cancer cell (NSCLC) line, PC-14/CDDP, were evaluated. The transplantability of the tumor xenografts to SCID mice was more than 90%. Tumor xenografts of H69/CDDP and PC-14/CDDP showed CDDP resistance during in vivo treatment. The novel anticancer agent 254-S showed only a partial effect on the growth of H69/CDDP and PC-14/CDDP while ormaplatin showed no cross resistance to CDDP. The in vivo results correlated well with the results of the in vitro MTT assay. In this in vivo sensitivity test, H69/CDDP and PC-14/CDDP were more sensitive to ormaplatin than its parental cell lines. In vivo sensitivity testing using SCID mice bearing transplanted CDDP-resistant tumors was shown to be useful for evaluating the effects of new anti-cancer drugs, especially those that might overcome CDDP resistance.

Characterization of an etoposide-resistant human ovarian cancer cell line.

Etoposide (VP-16) is one of the most important anticancer agents available and is used in many chemotherapeutic regimens. To characterize resistance to this drug, we established a VP-16-resistant human ovarian cancer cell line, SKOV3/VP, by continuous stepwise exposure of SKOV3 cells to VP-16. The degree of resistance to VP-16 of SKOV3/VP was about 25 times that of the parent cell line (SKOV3), and SKOV3/VP showed cross-resistance to teniposide, adriamycin, CPT-11, and vincristine. The accumulation of [3H]-VP-16 observed in SKOV3/VP cells was about half that seen in SKOV3 cells, and the accumulation of Adriamycin by this resistant cell line was also lower than that of its parent. Overexpression of neither the multidrug resistance gene mdr-1, the multidrug-resistance-associated protein (mrp) gene, nor P-glycoprotein was detected using reverse transcriptase-polymerase chain reaction analysis and flow cytometry with MRK-16, a monoclonal antibody against P-glycoprotein. The topoisomerase II activity of nuclear extracts from SKOV3/VP cells was lower than that from the parental cells, as was the amount of DNA topoisomerase II, demonstrated by immunoblotting. These results suggest that the mechanism responsible for the multidrug resistance of this cell line may be attributable to changes on its DNA topoisomerase II and to its reduced accumulation of the drugs as compared with the parental line SKOV3.

Intraduodenal capsaicin inhibits gastric migrating motor complex via an extrinsic neural reflex in conscious dogs.

The aim was to study the effect of intraduodenal capsaicin on interdigestive gastric contractions. Mongrel dogs were equipped with strain-gauge force transducers to measure gastroduodenal motility. The effects of intraduodenal capsaicin with or without pharmacological antagonists on spontaneous and motilin-induced interdigestive gastric contractions and on plasma motilin were studied in dogs with intact stomachs. The effect of intraduodenal capsaicin on gastric contractions was also studied in vagally denervated gastric (Heidenhain) pouch and vagally innervated antral pouch. Intraduodenal capsaicin inhibited spontaneous and motilin-induced gastric contractions. The spontaneous peak in plasma motilin was inhibited by intraduodenal capsaicin. The effect of intraduodenal capsaicin on motilin-induced gastric contractions was not affected by blockade of nitric oxide synthase, or by beta-adrenoceptor antagonist. Administration of alpha-adrenergic blocker inhibited basal interdigestive gastric motility. Intraduodenal capsaicin had no effect on contractions in the Heidenhain pouch but inhibited those in vagally innervated antral pouch. Duodenal afferent fibres stimulated by capsaicin inhibit gastric contractions via a nitric oxide-independent extrinsic neural reflex.

Treatment of severe ovarian hyperstimulation syndrome by ultrafiltration and reinfusion of ascitic fluid.

Two severe OHSS patients were treated by ultrafiltration and reinfusion of their ascitic fluid. Improvement of symptoms was marked after this treatment with no complications, and termination of pregnancy was avoided. Treatment of OHSS with this ultrafiltration and reinfusion method may help to resolve serious cases of OHSS and become a useful treatment for severe OHSS.

Colonization of microflora in mice: mucosal defense against luminal bacteria.

To investigate the pathogenesis of inflammatory bowel disease, it is critical to develop a system that uses simple and reproducible models for analyzing the "normal" mucosal defense mechanism. In the present study, germ-free mice were conventionalized by the oral administration of microorganisms prepared from the feces of genetically identical mice. Histological assessment and mucin characterization of small intestine and colon were then carried out. Histological findings in the gut were site-dependent and clearly time-dependent. Acute inflammation was most evident in the cecum. The cecal mucosa exhibited hyperplastic changes in epithelial cells, infiltration of polymorphonuclear cells, crypt abscesses, and epithelial projections on the epithelial surface 7 days after conventionalization. Some of the changes were similar to those seen in human ulcerative colitis. The histological findings in the conventionalized mice were comparable to those in specific pathogen-free mice after 28 days. Mucin histochemistry revealed that bacterial colonization altered the number of rectal goblet cells and the mucin composition in a time-dependent fashion. Although this model shares only some characteristics of human inflammatory bowel disease, it is unique in demonstrating the acquisition of mucosal defense. Understanding of this process is critical for the elucidation of inflammatory bowel disease pathogenesis.

Chronic diversion of bile to the urinary bladder induces pancreatic growth in dogs.

The aim of this study was to elucidate the mechanism of chronic biliary diversion and its effect on pancreatic growth. In the first part of the study, nine mongrel dogs underwent diversion of bile from the gastrointestinal tract by ligating the common bile duct and interposing a segment of jejunum between the gallbladder and the urinary bladder (cholecystojejunocystostomy [CJC]). Despite the loss of 7% of their body weight at 12 weeks after bilioenteric diversion, CJC dogs had significantly greater pancreatic wet weight than control dogs (51.2 +/- 2.2 g vs. 37.1 +/- 2.2 g). In the second part of the study, six other dogs underwent CJC. Twelve weeks later, bilioenteric continuity was restored by creating a cholecystojejunoduodenostomy (CJD). The dogs were given butter (3 g/kg) by mouth (prior to surgery, 12 weeks after CJC, and 4 weeks after CJD). Pancreatic excisional biopsy specimens were obtained at each operation and at autopsy. CJC induced more pancreatic RNA per milligram of weight (743 +/- 52, CJC; 579 +/- 44, prior to surgery, P <0.05 vs. CJC; 520 +/- 26 microg/100 mg tissue, CJD, P <0.01 vs. CJC), but not more DNA, and significantly higher basal plasma cholecystokinin levels and butter-stimulated cholecystokinin responses when compared with values prior to surgery or following CJD. We conclude that chronic biliary diversion induces pancreatic growth associated with hypersecretion of cholecystokinin in dogs.

Cross-resistance to antineoplastic agents in a human small-cell lung-cancer subline resistant to okadaic Acid.

We report on a human small cell lung cancer subline (H69/OA100) resistant to okadaic acid, an inhibitor of protein phosphatases. H69/OA100 showed cross-resistance to cis-diamminedichloroplatinum(II) (CDDP), adriamycin, and vinca alkaloids. Intracellular retention of adriamycin and CDDP in H69/OA100 was the same as those in H69. H69/OA100 was not shown to express MDR-1 by the reverse transcription polymerase chain reaction method. Expression level of mRNA of multidrug resistance-associated protein (MRP) in H69/OA100 was the same as that in H69. These data suggest that the mechanism of drug resistance in H69/OA100 might be due to a new mechanism of non-P-glycoprotein mediated multidrug resistance.

[Gastric emptying of semi-solid meal in normal man with special reference to relationship between plasma acetoaminophen concentrations and gastric emptying time].

Correlation between gastric emptying time measured by a radioisotope method and plasma acetoaminophen level was studied in 11 preoperative early gastric cancer in semisolid meal. A significant correlation was found between both gastric emptying half-time and plasma acetoaminophen level at 30, 45 minutes after ingestion of meal, and plasma acetoaminophen level at maximum. We conclude that acetoaminophen analysis is useful to measure gastric emptying at semisolid meal.

Management of small cell lung cancer in elderly.

BACKGROUND: There is increasing interest in the treatment of elderly small cell lung cancer (SCLC) patients, however the patients enrolled in clinical studies are strictly selected elderlies. MATERIALS AND METHODS: We retrospectively reviewed the treatment of consecutive unselected patients more than 70 years of age in whom SCLC were diagnosed over the past 22 years. RESULTS: In patients who received cisplatin-containing chemotherapy, response rate(RR) and mean survival time(MST) were 53.8% and 9 months. Whereas, in patients who were treated with carboplatin-containing chemotherapy, RR and MST were 72.0% and 7 months. The latter group included the patients whose doses of carboplatinum was reduced 20 to 50% in the first course of treatment. There was no difference found in RR, survival, and incidence of toxicity between these chemotherapy groups. CONCLUSION: Our results showed that in elderly SCLC patients, carboplatin could be substituted for cisplatin without apparent loss of therapeutic efficacy.

Suramin inhibits the phosphorylation and catalytic activity of DNA topoisomerase II in human lung cancer cells.

Suramin is a prototype of a new class of anticancer drugs. We investigated the action of suramin on the signal transduction pathways to DNA topoisomerase II (Topo II). Suramin showed a growth-inhibitory effect on a human lung cancer cell line (PC-9) with an IC50 of about 160 micrograms/ml. Suramin inhibited the catalytic activity of Topo II with an IC50 of about 100 micrograms/ml without stabilization of the cleavable complex of DNA and Topo II. Suramin decreased the phosphorylation of Topo II with an IC50 of 175 micrograms/ml, but did not change the degree of Topo II expression. These IC50 values for inhibition of catalytic activity and phosphorylation of Topo II were equivalent to the growth-inhibitory dose determined by tetrazolium dye assay. Phosphorylation of the tyrosine residues of Topo II was not changed by suramin. In the presence of okadaic acid, a potent inhibitor of serine/threonine protein phosphatase, suramin also decreased the phosphorylation of Topo II, suggesting that the drug did not act on the serine/threonine protein phosphatases inhibited by okadaic acid. Suramin also inhibited the protein kinase C (PKC) activity of PC-9 cells. These results suggest that suramin decreases the phosphorylation of Topo II mediated by PKC. This effect of suramin might cause the inhibition of Topo II activity resulting in the growth inhibition of tumor cells.

Effects of beta- and gamma-carboline derivatives of DNA topoisomerase activities.

beta-Carbolines, harman (1-methyl-9H-pyrido[3,4-b]indole) and norharman (9H-pyrido[3,4-b]indole) and gamma-carbolines, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) and 3-amino-4-methyl-5H-pyrido[4,3-b]indole (Trp-P-2), are present in cooked foods and cigarette smoke. We studied the effects of these heterocyclic amines on the activity of DNA topoisomerases. Trp-P-1 and Trp-P-2 inhibited topoisomerase I (topo I) activity with ED50 values of 1.48 and 1.55 micrograms/ml, respectively, in a relaxation assay. Harman and norharman inhibited topo I activity but with much higher ED50 values, 23.8 and 34.4 micrograms/ml, respectively. Trp-P-1 and Trp-P-2 also inhibited topoisomerase II (topo II) activity at about 50 micrograms/ml, in a decatenation assay. Harman and norharman showed a much lower inhibitory effect on topo II activity. None of these compounds stabilized the cleavable complex mediated by topo II. Trp-P-1 and Trp-P-2 intercalated into DNA at concentrations inhibitory to topoisomerases. We considered that the intercalation with DNA and the inhibition of DNA topoisomerases by heterocyclic amines might be partly related to their inhibition of DNA excision repair and their enhancing effect on UV- or chemically induced mutagenic activity.

The influence of age and health status on the serum alpha 1-acid glycoprotein level of conventional and specific pathogen-free pigs.

Serum alpha-1-acid glycoprotein (alpha 1AG) was measured in 212 Landrace White pigs between birth and finishing age. The alpha 1AG level of healthy pigs five to ten months of age was 338 +/- 79 micrograms/mL, and the upper normal limit in mature swine has been established as 500 micrograms/mL. In both specific pathogen-free (SPF) and conventional pigs, the alpha 1AG level within one day of birth was 14,263 +/- 2,393 micrograms/mL, 40 times the normal adult value, but rapidly decreased to 699 +/- 186 micrograms/mL by four weeks of age. In conventional pigs, alpha 1AG began to increase after four weeks, averaged 1,428 micrograms/mL by eight weeks, but gradually decreased to adult levels by 20 weeks of age. In comparison, alpha 1AG of SPF pigs was only 800 micrograms/mL at eight weeks and decreased more rapidly to normal by 16 weeks of age. The conventional pigs had a high incidence of clinical pneumonia and specific antibodies to Actinobacillus pleuropneumoniae and Mycoplasma hyopneumoniae at the age of eight weeks. As the clinical pneumonia disappeared, serum alpha 1AG level also gradually declined. In contrast, SPF pigs had little clinical illness, low alpha 1AG, and little serological evidence of microbial infection. Conventional pigs with nonrespiratory infections, encephalitis, or with hernias had increased alpha 1AG. While the very high alpha 1AG level of the neonatal pig may be due to genetic influences, increases later in life are likely in response to stimuli from its external environment. Monitoring of serum alpha 1AG in several herds aided in the recognition of disease processes and may have potential use in swine herd health management.

Characterization of serum alpha 1-acid glycoprotein in fetal and newborn calves during development.

Serum alpha 1-acid glycoprotein (alpha 1-AG) in bovine fetuses and newborn calves was characterized. Serum alpha 1-AG concentration increased during fetal development and neonatal stages. Mean +/- SD serum alpha 1-AG concentration reached a peak of 1,368 +/- 207 micrograms/ml immediately after birth, but thereafter gradually decreased to 249 +/- 100 micrograms/ml, similar to the normal adult bovine range. By use of isoelectric focusing of thin-layer gels, we detected 7 microheterogeneity bands ranging from pI 3.2 to 3.8 in adult bovine serum. Twelve bands ranging from pI 2.6 to 3.8 were found in 9-month fetuses and in neonates. The 5 most-acidic bands, which are absent in adult serum, ranged between pI 2.6 and 3.1 and decreased with maturation as band patterns assumed adult characteristics. By crossed affinity electrophoresis, alpha 1-AG of adult bovine serum was separated into 4 peaks according to its differential affinity to concanavalin A (conA). Seventy-five percent of the alpha 1-AG concentration was represented by peak 3 (P-3) and peak 4 (P-4), which had moderate or strong binding to conA. In contrast, fetal sera contained only peak 1 (P-1), which did not have conA-binding affinity. In neonatal sera, 4 peaks were recognized, of which P-1 comprised 70% of the total alpha 1-AG. Thereafter, with aging, percentage of P-3 and P-4 increased as band composition approached the normally expected adult pattern.

[The coexistence of pulmonary tuberculosis and thymoma a case report].

A 33-year-old male was admitted with complaints of cough, dysphagia, and swelling of face and upper extremities. Chest X-ray and CT scan revealed a large mediastinal mass and infiltrates in the right upper lobe. Percutaneous biopsy proved the mediastinal tumor as thymoma with cellular atypia. After irradiation, the tumor was surgically removed. Caseous epitheloid granulomas were found in the dissected mediastinal lymph nodes. AFB (Acid fast bacillus) stain of the patient's gastric fluid was positive for Mycobacterium. The coexistence of these two diseases was incidental, however, this case suggested that clinicians should perform careful evaluation of lung parenchyma as well as mediastinum on chest X-ray to identify occult diseases including pulmonary tuberculosis in patients with mediastinal mass lesion.