Spontaneous complete regression of a rectal cancer.

We report a unique case of a biopsy-proven rectal cancer exhibiting spontaneous complete regression in an extremely short period of 3 months. An 80-year-old man visited our hospital because of a positive fecal occult blood test. Colonoscopy showed a sessile polyp, about 25 mm in diameter, in the middle part of the rectum. Instead of endoscopic resection, two endoscopic biopsies were taken for histological evaluation, as an invasive cancer was endoscopically suspected.Well-differentiated invasive adenocarcinoma was revealed, and thus surgical resection was planned. At the second colonoscopy for endoscopic tattooing before surgery, the polyp was found to have unexpectedly developed into a flat lesion. Furthermore, the surgically removed specimen showed that the flat lesion had transformed to a depressed lesion, and surprisingly, no cancerous tissue was detected histologically.

Outbreak of Nocardia farcinica infection with the same pattern in randomly amplified polymorphic DNA analysis.

We experienced three cases of nocardiosis by Nocardia farcinica in the same ward within a six-month period. The result of gene analysis by randomly amplified polymorphic DNA gave the same pattern. Thus, these three cases were considered to be caused by the same strain of N. farcinica, implying the presence of nosocomial infection.

The cloning of eggplant seedling cDNAs encoding proteins from a novel cytochrome P-450 family (CYP76).

Using a CYP75 cDNA as a probe, we have cloned and sequenced two closely related cDNAs from eggplant seedlings, which encode typical cytochrome P-450 (P450) proteins. A database search revealed that the predicted proteins share less than 40% amino acid homology with other P450 proteins, which suggests that they form a novel P450 gene family (CYP76).

Mucosa-associated bacteria in ulcerative colitis before and after antibiotic combination therapy.

BACKGROUND: We proposed that Fusobacterium varium is one of the causative agents in ulcerative colitis. AIM: To examine the efficacy of antibiotic combination therapy against F. varium and to investigate the mucosa-associated bacteria before and after the therapy using a new molecular approach. METHODS: Twenty patients with ulcerative colitis were randomly assigned into the antibiotic treatment group (amoxicillin, tetracycline and metronidazole for 2 weeks) and no-antibiotics group. Clinical assessment, colonoscopic and histological evaluations were performed at 0 and 3-5 months after the treatment. DNA from mucosal bacteria was isolated from biopsy specimens. We investigated the mucosa-associated bacterial components by terminal restriction fragment length polymorphism with the restriction enzyme HhaI and MspI, and quantified the change in the number of bacteria by real-time polymerase chain reaction. Immunohistochemical detection of F. varium in biopsy specimens was also performed. RESULTS: After the treatment, the clinical assessment, colonoscopic and histological scores improved in the antibiotic group compared with the control group. Three peaks of terminal restriction fragment length polymorphism decreased after treatment only in the antibiotic group. Eubacterium rectale, Dorea formicigenerans, Clostridium clostridioforme and F. varium were included in these peaks. Based on the real-time polymerase chain reaction study, only F. varium was significantly reduced after treatment. In the immunostaining, post-treatment scores in treatment group were significantly lower than that in control group. CONCLUSIONS: Antibiotics combination therapy was effective for ulcerative colitis. The number of mucosa-associated F. varium significantly decreased after the treatment.

Atrial natriuretic peptide in patients with the syndrome of inappropriate antidiuretic hormone secretion and with diabetes insipidus.

To examine a possible role for atrial natriuretic peptide (ANP) in water and sodium metabolism disturbances associated with abnormal vasopressin (AVP) secretion, we measured plasma ANP concentrations in 15 patients with the syndrome of inappropriate antidiuretic hormone secretion (SIADH) and in 17 patients with central diabetes insipidus (DI). The mean plasma ANP concentration (30.2 +/- 10.4 pmol/L) in SIADH patients who had hyponatremia, plasma hypoosmolality, hyperosmolar urinary compared to plasma sodium levels, and increased plasma AVP levels relative to plasma osmolality was significantly higher than that in normal subjects (12.6 +/- 4.9 pmol/L), although there was a considerable individual variation in plasma ANP ranging from normal to clearly elevated levels (15.1-47.0 pmol/L). When hyponatremia was corrected by water restriction or demeclocycline administration, plasma ANP levels decreased significantly and fell into the normal range (12.5 +/- 4.3 pmol/L). DI patients who complained of polyuria and polydipsia and had hypoosmolar urine, normal or elevated plasma sodium concentrations, and decreased plasma AVP levels relative to plasma osmolality, on the other hand, had a significantly lower mean plasma ANP level (7.6 +/- 2.9 pmol/L) than normal subjects. There was, again, a considerable overlap between plasma ANP levels in individual DI patients (4.2-13.9 pmol/L) and those in normal subjects. Treatment with 1-desamino-8-D-arginine vasopressin resulted in a significant increase in the mean plasma ANP level (18.6 +/- 8.0 pmol/L). There were no significant correlations between plasma ANP and AVP levels in either group of patients. The results indicate that ANP secretion is modulated by changes in plasma volume consequent to abnormal AVP secretion, which may have a pathophysiological significance in maintaining volume homeostasis.

cDNAs sequences encoding cytochrome P450 (CYP71 family) from eggplant seedlings.

Three cytochrome P450 (P450) cDNAs were isolated from an eggplant hypocotyl cDNA library using eggplant CYP75 cDNA as a probe. These cDNAs have greater than 65% identity in their amino acid sequences, indicating that they belong to the same family. Comparison of these with P450 proteins from other sources showed that the protein with the greatest degree of homology is CYP71, isolated from avocado fruits (approximately 48%). We concluded that they are novel members of the CYP71 gene family (CYP71A2, 3 and 4). We have examined the level of mRNA transcripts from the CYP71 family in eggplant hypocotyl tissues and petunia flower buds, and found that the level of transcripts is developmentally regulated in the flower bud.

Congenital muscular dystrophy: Clinical and pathologic study of 50 patients with the classical (Occidental) merosin-positive form.

We studied 50 patients with the merosin-positive form of congenital muscular dystrophy (MP-CMD) clinically and pathologically. The frequency of MP-CMD in our laboratory was approximately one-half that of the Fukuyama type and one-sixth that of Duchenne muscular dystrophy. The early signs of MP-CMD included decreased fetal movement during pregnancy (14%) and poor suck (42%), floppiness (30%), and respiratory difficulty (16%) in early infancy. Eighty-six percent of the patients had delayed motor development. Ninety-two percent of the patients followed beyond age 4 years had learned to walk. The disease was relatively slowly progressive, except in six patients who rapidly lost ambulation. Almost all patients had normal IQ, except four who were mildly to moderately retarded. Of the patients examined by cranial CT/MRI, 24% showed cerebral atrophy and 11% had areas of white matter lucency. Muscle biopsy results in those younger than 5 years showed mild dystrophic changes consisting of variation in fiber size and scattered necrotic and regenerating fibers. In older children, there were additional chronic dystrophic changes, including fiber splitting (32%), moth-eaten appearance (32%), marked fatty replacement (46%), and abnormal fiber type distribution (59%). The manifestations of MP-CMD were generally milder and more slowly progressive than those of the Fukuyama type and merosin-negative form of congenital muscular dystrophy.

MTM1 gene mutations in Japanese patients with the severe infantile form of myotubular myopathy.

The severe infantile form of myotubular myopathy is a fatal muscle disease that predominantly affects male infants and is characterized by severe weakness and hypotonia from birth. X-linked myotubular myopathy was found to be associated with mutations in the MTM1 gene in Xq28 encoding the putative tyrosine phosphatase, myotubularin. We screened the MTM1 gene for mutations in seven Japanese patients (six males and one female) who had the diagnosis of severe infantile form of myotubular myopathy. We found five mutations, including three novel mutations based on sequence analysis of RT-PCR fragments covering the entire open reading frame. Two patients (one male and one female), who had similar clinicopathologic features, did not have any mutation in the MTM1 gene open reading frame, suggesting that they may have had an autosomal recessive disease.

Heterogeneity of presynaptic muscarinic receptors involved in modulation of transmitter release.

In order to extend the characterization of muscarinic receptors at presynaptic sites their inhibitory effect on the stimulation-evoked release of [3H]noradrenaline and [3H]acetylcholine from different axon terminals was studied and the dissociation constants and potencies of different antagonists were estimated, in guinea-pig and rat. While oxotremorine reduced the release of [3H]acetylcholine and [3H]-noradrenaline in a concentration-dependent manner from different release sites (Auerbach plexus, noradrenergic neurons in the right atrium, cerebral cortex), McN-A 343, an M1 receptor agonist, enhanced their release evoked by field stimulation. When the inhibitory effect of oxotremorine on transmitter release was studied, pancuronium, pirenzepine and atropine were competitive antagonists of presynaptic muscarinic receptors located on the noradrenergic axon terminals of the atrium. While atropine and pirenzepine inhibited the muscarinic receptors of cholinergic axon terminals in the Auerbach plexus, pancuronium and gallamine had a very low affinity. Significant differences were found in the affinity constants of antagonists for muscarinic receptors located in the cholinergic axon terminals of Auerbach plexus and cerebral cortex, and noradrenergic axon terminals of the atrium. While atropine and pirenzepine exerted similar effects on these presynaptic sites, pancuronium, gallamine and (11-(2-[diethylamino)-methyl)-1-piperidinyl)acetyl)-5, 11-dihydro-6(1-pyrido(2,3-b)(1,4)-benzodiazepin-6-on) were much more effective on muscarinic receptors controlling acetylcholine release from the cerebral cortex and noradrenaline release from the heart. There was more than 100-fold (2.0 pA2 units) difference in affinities of these antagonists.(ABSTRACT TRUNCATED AT 250 WORDS)

Prognostic value of tissue inhibitor of matrix metalloproteinase-1 in plasma of patients with gastric cancer.

Tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in plasma has been reported to be related to disease progression in patients with gastric cancer. However, the prognostic significance of plasma TIMP-1 concentrations has not been clarified. Concentrations of TIMP-1 protein were measured by enzyme-linked immuno-sorbent assay in plasma samples of 147 preoperative patients who subsequently underwent gastric resection, and prognosis was compared. The cut-off value of plasma TIMP-1 concentrations was defined as 112.5 ng/ml, referring to the TIMP-1 levels in patients with intramucosal gastric cancer. Twenty-nine out of 147 patients had higher plasma TIMP-1 levels than the cut off value. When the patients were divided into those with elevated values and those with normal TIMP-1, such parameters as age, serosal invasion, metastases to lymph nodes, peritoneum, and liver, lymphatic invasion, curability, and stage were significantly different between the two. By univariate analysis of the factors affecting survival, macroscopic type, histology, serosal invasion, metastasis to lymph node, peritoneum, and liver, vessel invasions, curability, and plasma TIMP-1 were significant. However, multivariate analysis revealed that TIMP-1 was the only significant factor. In patients with gastric cancer, plasma TIMP-1 seem to be an independent and most powerful prognosticator for the survival.

Plasma concentrations of VEGF and bFGF in patients with gastric carcinoma.

We examined plasma levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in 54 patients with gastric carcinoma. Postoperative survival was significantly poorer in patients with plasma VEGF levels more than 10.0 pg/ml at the time of surgery. By an univariate analysis of the factors affecting survival, serosal invasion, lymph node metastasis, peritoneal dissemination, lymphatic vessel invasion, curability, and VEGF proteins were significant. By a multivariate analysis only VEGF levels and curability remained significant. Patients with recurrent disease, including liver metastasis, had significantly higher plasma VEGF concentrations than those with resectable primary tumors. VEGF, not bFGF, may serve as an independent prognosticator and a sensitive indicator for liver recurrence in patients with gastric carcinoma.

Inhibition of proliferation of gastric epithelial cells by a cyclooxygenase 2 inhibitor, JTE522, is also mediated by a PGE2-independent pathway.

BACKGROUND: Cyclooxygenase-2 (COX-2) is one of the rate-limiting enzymes for prostaglandin synthesis from arachidonic acid. Although it is known that inhibition of cyclooxygenase activity delays ulcer healing, the regulatory relationship between COX-2 and its metabolites in gastric epithelial cell proliferation is not well known. AIM: To investigate whether COX-2 has an effect on gastric mucosal cell proliferation and further studied whether such effect is mediated only by prostaglandin E2 (PGE2), a representative metabolite of arachidonates in the gastric mucosa. METHODS: Artificial wounds of defined area size were created on complete monolayer cell sheets of isolated rat gastric epithelial cells and rat gastric cell line RGM1 under the addition of arachidonic acid or a COX-2 selective inhibitor, JTE522. Repair of wounds was assessed by monitoring wound size, with cell proliferation detected using 5-bromodeoxyuridine staining. Quantity of secreted PGE2 was measured by enzyme immunoassay. RESULTS: Stimulation of foetal calf serum increased the expression of COX-2 protein and inhibition of COX-2 retarded wound healing with reduction of cell proliferation. Arachidonic acid increased PGE2 production and accelerated restoration. Combination of JTE522 and arachidonic acid resulted in a marked retardation of wound healing compared to the control, but JTE522 did not completely suppress the increase in cellular PGE2 content following the addition of arachidonate. CONCLUSIONS: The difference in the effects of JTE522 on PGE2 production and on wound healing suggest that the involvement of COX-2 in gastric epithelial cell proliferation is not mediated solely by PGE2.

Monomer-dimer equilibria of a Bence Jones protein and its variable fragment.

The circular dichroic (CD) spectra of a type lambda Bence Jones protein (Tod), its variable (VL) fragment, and the constant (CL) fragment of a type lambda protein (Nag) were measured under various conditions. In the pH region from 5.5 to 7.5, the CD spectra of Tod protein with intact interchain disulfide bond (L(SS)) and and CL did not change with pH, while the spectra of Tod protein in which the interchain disulfide bond had been reduced and alkylated (L(RA)) and VL did not change with pH. The dimerization reactions of L(RA) and VL were studied by following the CD change with protein concentration. The CD spectrum of CL did not change with the protein concentration. The dimerization constant for L(RA) was 4 X 10(4) M-1 at at pH 7.5 and 25 degrees C, which was smaller than that for VL (1 X 10(5) M-1). The ellipticity at 278 nm for the L(RA) dimer was different from that for the L(SS) dimer and changed with pH. These findings indicate that the L(RA) dimer and L(SS) dimer have different conformations. The differences in the conformation and L-L interaction between the L(RA) dimer and L(SS) dimer are discussed on the basis of the conformations of VL and CL and the interactions between the paired domains.

Effect of dithiothreitol on activity and protein structure of human urine urokinase.

Human urine urokinase [EC 3.4.21.31] was found to be inactivated by dithiothreitol (DTT) much more severely than by 2-mercaptoethanol at the same concentration on the basis of -SH groups. Removal of DTT by dialysis restored the activities of esterase toward acetyl-glycyl-L-lysine methyl ester, plasminogen activation, and amidase toward 7-(glutaryl-glycyl-L-arginine-amido)-4-methyl coumarin. But the restoration of amidase activity was much less than that of esterase activity. The addition of DTT mediated the conversion of high molecular weight urokinase to low molecular weight urokinase, releasing several peptides. This suggests that the urokinase consists of several polypeptides linked by disulfide bonds. The molecular weight of urokinase produced with DTT was smaller than that of low molecular weight urokinase obtained by autodigestion of high molecular weight urokinase. The autodigestion was also accompanied by liberation of some peptides. But, those peptides released on autodigestion of high molecular weight urokinase were different from those appearing in the presence of DTT.

Uropepsinogen in human urine: its protein nature, activation and enzymatic properties of activated enzyme.

Uropepsinogen (UPG) in human urine was highly purified by chromatography on columns of DEAE-lignocellulose, elastin-celite, etc. The purified UPG was composed of two electrophoretically distinguishable components (UPG I and UPG II), and they were isolated in the pure state, respectively. UPG I and UPG II were the same in molecular weight (3.9 X 10(4)) and in N-terminal (leucine) and C-terminal (alanine) amino acid residues. Also, they were quite similar in amino acid composition. They were activated to uropepsin (UP I and UP II, molecular weight, 3.3 X 10(4), respectively), but the activated enzymes were the same in the various properties examined, suggesting that the difference between UPG I and UPG II is due to only a minor change in the peptide segment, perhaps by deamidation. The activation of UPG I and UPG II occurred at acid pHs, the best pH being at 2.0. Both the proenzymes previously incubated with pepstatin at pH 6.8, however, were not activated even in the following incubation at acid sides. The results obtained are discussed in regard to the origin of the proenzyme and its properties before and after activation.

The acute effects of prostaglandin E1 on the pulmonary circulation and oxygen delivery in patients with the adult respiratory distress syndrome.

Prostaglandin E1 was administered intravenously to 10 patients who had the adult respiratory distress syndrome associated with severe infection in order to investigate its hemodynamic effects. Infusion of PGE1 significantly decreased the mean pulmonary arterial pressure, mean systemic arterial pressure, pulmonary vascular resistance and systemic vascular resistance, and increased the cardiac index, oxygen delivery and oxygen consumption. No significant difference was noted in the intrapulmonary shunt fraction. These results indicate that administration of PGE1 improves pulmonary hemodynamics and tissue oxygenation in patients with acute respiratory distress syndrome, by reducing right ventricular afterload and increasing the cardiac index.

Insufficient diagnostic accuracy of imported serological kits for Helicobacter pylori infection in Japanese population.

Although there are many reports of the high diagnostic accuracy of commercially available serologic kits for Helicobacter pylori infection in Western countries, they rarely has been investigated in oriental population. Accordingly we examined their usefulness in 492 Japanese patients with dyspeptic symptoms. Diagnostic accuracy of 4 imported serologic kits (HEL-p TEST, HM CAP, G.A.P IgG, Helico G2) was investigated using the (13)C-urea breath test as the gold standard. When intermediate results were excluded, the sensitivity, specificity and accuracy of these serologic tests ranged from 88.6% to 97.8%, 67.9% to 85.9%, and 87.9% to 91.4%, respectively, which were comparable with reported median accuracy in the Western population. However, there were many intermediate results in these tests, ranging from 5.3% to 23.0%. Their usefulness seemed to be limited in our patient population because of the large number of intermediate results.

Adjuvant radiotherapy after complete resection of thymoma.

Seventy patients were studied after undergoing complete resection of thymoma to determine the effect of postoperative adjuvant mediastinal radiotherapy on prognosis, with regard to clinical stage, histological type, and pleural factor. Pleural factor was defined as follows: p0, no adhesion to the mediastinal pleura; p1, fibrous adhesion to the mediastinal pleura without microscopic invasion; and p2, microscopic invasion of the mediastinal pleura. Recurrence of thymoma after complete resection was observed in 13 patients, 12 (92%) with pleural dissemination, 6 (46%) with local recurrence, and 2 (15%) with distant metastasis (types of recurrence are overlapping). In stage I and stage II p0 patients, no recurrence was observed, regardless of mediastinal radiotherapy. Whereas mediastinal irradiation completely prevented recurrence in stage II p1 patients, 4 (36.4%) nonirradiated stage II p1 patients experienced recurrence. In stage II p2 patients, 75% had pleural dissemination even after radiotherapy. A high incidence of recurrence was also observed in stage III, nonirradiated (25%) and irradiated (30%) patients. The results suggest that mediastinal irradiation for stage I and II p0 patients is not always necessary, and that therapy for stage II p1 is essential and also expected to decrease the recurrence rate. On the other hand, in stage II p2 and stage III thymomas, mediastinal irradiation is not sufficient to prevent pleural recurrence even after complete resection. Our classification based on pleural factor is useful for better selection of appropriate postoperative treatment for thymoma patients.

Management of chylothorax after pulmonary resection.

BACKGROUND: Conservative management with intrapleural drainage and total parenteral nutrition (TPN) has been the first choice of treatment for postoperative chylothorax. With this approach, however, it usually takes several weeks for the chylothorax to resolve and it is sometimes unsuccessful. In this study, we reviewed seven patients who had chylothorax develop after pulmonary resection for primary carcinoma of the lung. STUDY DESIGN: The patients were treated according to a "one-week trial" that consisted of one week of observation with intrapleural drainage and maximum parenteral nutritional support followed by operative intervention if the effect of the conservative therapy was not adequate. When the chylous leak was decreased to less than 100 mL/day or less than 15 percent of the maximum daily drainage volume after the "one-week trial," the conservative management was continued for two more weeks. After observation for three weeks, oral intake was begun and a final evaluation of the treatment was made. RESULTS: One patient did not consent to the "one-week trial" and underwent operative treatment on the third postoperative day. Two patients had chylous leaks less than 100 mL/day or less than 15 percent of the maximum daily chylous leak after one week observation. Conservative management with TPN was continued in these patients for two more weeks and operation was performed in one on the 20th day and in the other on the 22nd postoperative day. The remaining four patients underwent operative treatment on the seventh or eighth postoperative day. All of the operations for chylothorax were successful, and chest tubes were removed promptly. These results show that operative management of chylothorax was reliable and safe. The "one-week trial," however, offered few advantages in determining the therapeutic strategy for postoperative chylothorax.

Intrapulmonary metastasis of lung cancer: soft x-ray investigation of inflated and fixed lung.

BACKGROUND: Intrapulmonary metastasis (IPM) of lung cancer is thought to be an important factor influencing patient prognosis. It is not easy to detect a small IPM by preoperative examination and sometimes even by postoperative pathologic investigation. We applied soft x-ray investigation to inflated and fixed lungs for the detection of IPM. STUDY DESIGN: From 1990 to 1992, 75 patients with lung cancer who had no metastatic lesions on preoperative whole CT, MRI, and technetium-99m bone scintigram examinations underwent lung resection. The resected lungs were fixed in an inflated condition, sliced at the corresponding CT levels into 10-mm-thick sections, and submitted for soft x-ray examination. When an accessory nodular shadow(s) was detected on the soft x-ray images, the size of the nodule and its distance from the primary tumor were measured. RESULTS: In 23 of the 75 patients, accessory nodular shadows were detected on the soft x-ray images. Six nodules in 6 patients proved to be IPM, 2 of which were also detected by postoperative macroscopic examination. Another 2 microscopic IPM were found only by postoperative pathologic examination. The total detection rate of IPM was 10.7% (8 of 75 patients) in this series. The detection rate of IPM at our institute was 5.4% before this study (1979 to 1989). The mean diameter of the IPM detected by the soft x-ray method was 2.8 +/- 1.5 mm, and this was significantly smaller than that of the macroscopically detected nodules (7.2 +/- 3.2 mm). CONCLUSIONS: Our data show that soft x-ray investigation is an effective procedure to detect relatively small intrapulmonary metastatic nodules and will contribute to precise postoperative staging of patients with lung cancer.