Secretion of human intracellular aspartic proteinase cathepsin E expressed in the methylotrophic yeast, Pichia pastoris and characterization of produced recombinant cathepsin E.

The human gastric cathepsin E (CTSE), a dimeric aspartic proteinase, was expressed in the methylotrophic yeast Pichia pastoris by placing the CTSE cDNA under the control of the methanol inducible alcohol oxidase promoter. The human CTSE expressed in P. pastoris was secreted into the culture medium as an active enzyme directed by its native signal sequence despite its intracellular localization in mammalian cells. The time course analysis of the culture supernatant of the P. pastoris transformant expressing human CTSE revealed that the recombinant human CTSE was secreted as a 90 kDa molecule and then converted via an 84 kDa intermediate to an 82 kDa mature molecule. A large-scale culture of the transformant was performed in a high cell density fermentor and the recombinant human CTSE was highly purified from the culture supernatant. The purified recombinant cathepsin E had the molecular mass of 82 kDa with the amino-terminal sequence starting with Ile37 of the sequence deduced from its cDNA sequence, suggesting that the human cathepsin E was accumulated in the culture supernatant as mature dimeric enzyme. The result of endoglycosidase-H digestion followed by Western blot analysis of the purified recombinant cathepsin E suggested that the human cathepsin E expressed in P. pastoris received N-linked high-mannose type glycosylation. The enzymatic properties of the recombinant enzyme were comparable to those of natural human CTSE.

Meiotic behavior and linkage relationships in the secondarily homothallic fungus Agaricus bisporus.

This study followed the transmission of 64 segregating genetic markers to 52 haploid offspring, obtained from both homokaryotic and heterokaryotic meiospores, of a cross (AG 93b) of Agaricus bisporus, the commonly cultivated "button mushroom." The electrophoretic karyotypes of the AG 93b component nuclei were determined concurrently (n = 13). Eleven distinct linkage groups were identified by two-point analysis. DNA-DNA hybridization showed that nine of these corresponded to unique chromosome-sized DNAs. Two other chromosomal DNAs were marked with nonsegregating markers, including the rDNA repeat. Two remaining chromosomes remained unmarked but hybridized to repeated-sequence probes. Cross 93b had an essentially conventional meiosis in which both independent assortment and joint segregation of markers occurred, but in which crossing over was infrequent over much of the mapped genome. The 48 homokaryotic spore-offspring had overall crossover frequencies that were similar to, but possibly slightly less than, those of three homokaryon constituents of heterokaryotic spore-offspring. These daa provide support for our earlier cytogenetic model of sporogenesis in A. bisporus, that explains why heterokaryotic spore-offspring usually appear to exhibit no recombination. No evidence favoring an alternative, mitotic model of sporogenesis was found. The resulting genetic map appears to survey the genome extensively and for the first time permits localization of loci determining economically important traits in this fungal crop species. Large differences in the vigor of homokaryotic offspring were correlated with the inheritance of certain chromosome segments and were also often associated with significant departures from Mendelian segregation ratios.

Susceptibility of Helicobacter pylori isolates to metronidazole, clarithromycin and amoxycillin in vitro and in clinical treatment in Japan.

BACKGROUND: Primary and acquired resistance to antibiotics is an important factor in determining the reason for treatment failure in Helicobacter pylori infection. We examined the relationship between the susceptibility of H. pylori isolates and the efficacy of chemotherapy. METHODS: The minimal inhibitory concentrations (MICs) of metronidazole (MNZ), clarithromycin (CLAR) and amoxycillin (AMOX) of 320 H. pylori pre-treatment isolates were determined by the agar dilution method. In 290 patients with peptic ulcers. H. pylori infection was treated by dual or triple combination therapies for 2 weeks: one proton pump inhibitor (30 mg/day lansoprazole or 20 mg/day omeprazole) and one or two antibiotics (500 mg AMOX, 200 mg CLAR or 250 mg MNZ twice a day). MICs were also determined after the treatment failure. RESULTS: Among the drugs tested, for MNZ and CLAR, 8.1% and 9.1% of the isolates, respectively, were resistant, while no isolate was resistant to AMOX. After unsuccessful treatment using MNZ and CLAR, 66.7% and 70.61% of the isolates changed from sensitive to resistant, respectively. All isolates were sensitive to AMOX after treatment failure. CONCLUSIONS: The failure of the H. pylori treatment results in the induction of resistance to CLAR and/or MNZ. Regimens with a high cure rate should be used in order to prevent the generation of acquired resistance to antibiotics.

The effect of Helicobacter pylori eradication therapy on gastric antral myoelectrical activity and gastric emptying in patients with non-ulcer dyspepsia.

BACKGROUND: Dysmotility of the gastroduodenal region and delayed gastric emptying have been considered to play roles in non-ulcer dyspepsia (NUD). Helicobacter pylori-induced inflammation of the gastric mucosa may affect gastric motility. AIM: To evaluate the effects of H. pylori eradication therapy on gastrointestinal motility and symptoms in NUD patients. METHODS: : Forty-six NUD patients were examined for gastric emptying, antral myoelectrical activity, H. pylori infection, and symptom scores. In H. pylori-positive NUD patients, gastric emptying, antral myoelectrical activity, and symptom scores were also analysed 2 months after cure of H. pylori infection. RESULTS: Sixty-seven per cent of NUD patients were H. pylori-positive. Both abnormal gastric emptying and antral myoelectrical activity were observed in NUD patients. H. pylori-positive NUD patients were divided into three groups according to their gastric emptying: the delayed group, the normal group, and the rapid group. In the delayed and rapid gastric emptying groups, the emptying and symptom scores were improved significantly by eradication. There was no improvement in symptom scores in the normal gastric emptying NUD group by the eradication therapy. CONCLUSIONS: Disturbed gastric emptying and antral myoelectrical activity play roles in NUD. H. pylori-induced disturbed gastric emptying may cause some NUD symptoms. Gastric emptying and symptom scores are improved by H. pylori eradication therapy in NUD patients with disturbed gastric emptying; H. pylori eradication therapy is effective in H. pylori-positive NUD patients with disturbed gastric emptying.

The effect of helicobacter pylori eradication therapy on gastric antral myoelectrical activity and gastric emptying in patients with non-ulcer dyspepsia.

BACKGROUND: Dysmotility of the gastroduodenal region and delayed gastric emptying have been considered to play roles in non-ulcer dyspepsia. In addition, it has been reported that Helicobacter pylori induced inflammation of the gastric mucosa may affect gastric motility. AIM: To evaluate the effects of H. pylori eradication therapy on gastrointestinal motility and symptoms in non-ulcer dyspepsia patients. METHODS: A total of 46 non-ulcer dyspepsia patients were examined for gastric emptying, antral myoelectrical activity, H. pylori infection, and symptom scores. In H. pylori-positive non-ulcer dyspepsia patients, gastric emptying, antral myoelectrical activity, and symptom scores were also analysed 2 months after being cured of H. pylori infection. RESULTS: A total of 67.4% of the non-ulcer dyspepsia patients were H. pylori-positive. Both abnormal gastric emptying and antral myoelectrical activity were observed in non-ulcer dyspepsia patients. H. pylori-positive non-ulcer dyspepsia patients were divided into three groups according to their gastric emptying: the delayed gastric emptying group, the normal gastric emptying group, and the rapid gastric emptying group. In the delayed and rapid gastric emptying groups, the gastric emptying and symptom scores were improved significantly by the eradication therapy. However, there was no improvement in symptom scores in the normal gastric emptying non-ulcer dyspepsia group by the eradication therapy. CONCLUSIONS: Disturbed gastric emptying and antral myoelectrical activity play roles in non-ulcer dyspepsia. Helicobacter pylori infection, inducing disturbed gastric emptying, may cause some non-ulcer dyspepsia symptoms. Gastric emptying and symptom scores are improved by H. pylori eradication therapy in non-ulcer dyspepsia patients with disturbed gastric emptying. H. pylori eradication therapy is effective in H. pylori-positive non-ulcer dyspepsia patients with disturbed gastric emptying.

Helicobacter pylori an chronic atrophic gastritis.

Phenol red dye spraying endoscopy, in addition to a biopsy study, was employed for the determination of Helicobacter pylori distribution on the gastric mucosa of 108 patients with no gross gastric findings. The sensitivity of this method was 100%, and its specificity was 84.6%. In patients with a C0 pattern of the fundic-pyloric (F-P) border, the H. pylori-positive rate was only 21%, but this increased with the spread of atrophic mucosa, to approximately 90%. However, in patients with an O3 pattern, H. pylori could not be found. In addition, in 24 (80.0%) of 30 patients with no glandular atrophy, this organism was not found, while, in contrast, the organism was demonstrated in 54 (85.8%) of 63 patients with a slight degree of glandular atrophy. In patients with severe mucosal atrophy accompanying intestinal metaplasia, however, the organism was not found. One-half of the patients examined were followed up, using this technique. In those who were H. pylori-negative, glandular atrophy remained unchanged, while advanced glandular atrophy was seen in 10 (28.6%) of 35 H. pylori-positive patients. A cephalad shift of the F-P border was observed in 20 (57.1%) of these 35 H. pylori-positive patients, while in those who were negative, the F-P border remained unchanged, with one exception.

A proton pump inhibitor, E3810, has antibacterial activity through binding to Helicobacter pylori.

Helicobacter pylori infection is causally related to atrophic gastritis, and it may also be associated with peptic ulcer and gastric carcinoma. Eradication of H.pylori is recommended in patients with such diseases, especially in those with peptic ulcer. A new potent proton pump inhibitor, E3810, had an antibacterial effect on H. pylori, as has been reported for omeprazole and lansoprazole, two other proton pump inhibitors. The minimum inhibitory concentration of E3810 was 1.57-3.13 micrograms/ml, lower than that of omeprazole or lansoprazole. To clarify the mechanism of the antibacterial effect of E3810, we analyzed the characteristics of E3810 binding to H. pylori. Scatchard plot analysis of this binding showed a curvilinear profile, indicating the presence of several molecules with different affinities to E3810 on H. pylori. The binding capacity of E3810 to H. pylori was calculated to be about 2 x 10(6) sites/cell. These results suggested that E3810 has an antibacterial effect against H. pylori and that the effect may be mediated through direct binding to H. pylori.

Evaluation of endoscopic 13C-urea breath test for assessment of Helicobacter pylori eradication.

To minimize the inaccuracies of the diagnostic methods for Helicobacter pylori infection, we developed a new diagnostic method and called it the endoscopic 13C-urea breath test (EUBT). We evaluated the accuracy of EUBT for detecting this infection and assessing its eradication. EUBT was conducted on 267 patients with gastroduodenal disease. After the collection of a baseline breath sample, gastroduodenal endoscopy was performed. A 20-ml aliquot of a solution containing 100 mg of 13C-urea was sprayed over the entire gastric mucosa via the endoscope. A breath sample was then collected 15 min after spraying. The content of 13CO2 was measured in the breath samples by ratio mass spectrometry. Two biopsy specimens each from the antrum and the middle corpus were obtained for culture and histology. The EUBT cutoff level was 1.4%. The sensitivity and specificity of EUBT for the diagnosis of H. pylori infection were both 98.2%, and for assessment of the eradication they were 100% and 98.9%, respectively. EUBT is an accurate method not only for the diagnosis of H. pylori infection but also for assessment of its eradication.

Helicobacter pylori-negative gastric and duodenal ulcers.

It is unclear whether Helicobacter pylori infection is essential to the development of peptic ulcers. In this study, we examined the rates of H. pylori-negativity among patients with peptic ulcers. We also attempted to clarify the characteristics of H. pylori-negative peptic ulcers to throw light on the pathogenesis of peptic ulcers. The study included 215 consecutive patients with gastric ulcers (GUs) and 120 consecutive patients with duodenal ulcers (DUs). After routine endoscopic examination and phenol red dye endoscopy, forceps biopsies were performed for culture, histology, and the rapid urease test. A patient was considered H. pylori-negative when the serum anti-H. pylori IgG and the three tests on biopsied specimens were all negative. H. pylori-negative rates were 3.2% in the patients with GUs and 1.7% in the patients with DUs. Lack of atrophy of the gastric mucosa was significantly more common in the H. pylori-negative patients with GUs. A history of ulcer disease was less common and antral ulcers were more common in H. pylori-negative GU patients, but not significantly so. As the urea breath test had not been performed, the possibility of a false-negative result cannot be completely ruled out, but we believe that the H. pylori-negative rate in our study is more reliable than these rates in previous reports, because we visualized H. pylori distribution by phenol red dye endoscopy to avoid false-negative results in biopsies, and we used both biopsy and serum anti-H. pylori IgG findings to establish an H. pylori-negative diagnosis. Since H. pylori-negative peptic ulcers certainly exist, H. pylori infection is thought not to be essential to the development of peptic ulcers. There were few differences between the characteristics of H. pylori-negative and H. pylori-positive peptic ulcers in our study. A large-scale study is required to clarify the characteristics of H. pylori-negative peptic ulcers.

Vacuolating cytotoxin production by Helicobacter pylori isolates from peptic ulcer, atrophic gastritis and gastric carcinoma patients.

OBJECTIVE: We investigated the production of vacuolating cytotoxin by Helicobacter pylori isolates from patients with peptic ulcer, atrophic gastritis or gastric carcinoma in order to examine the pathophysiological significance of vacuolating cytotoxin in these diseases. METHODS: H. pylori was isolated from 18 patients (five with peptic ulcers, seven with atrophic gastritis and six with gastric carcinoma). Culture supernatants of H. pylori isolates, concentrated 20-fold, were serially diluted and then analyzed for cytotoxin activity semi-quantitatively using A431 cells as indicator cells. The relative activity of vacuolating cytotoxin was defined according to the maximum dilution. RESULTS: Cytotoxin production was observed in two out of five, six out of seven and six out of six isolates from peptic ulcer, atrophic gastritis and gastric carcinoma patients, respectively. The mean relative activity was calculated as 0.80, 2.71 and 2.50 in CONCLUSION: These results suggest that vacuolating cytotoxin-producing H. pylori is strongly associated with both atrophic gastritis and gastric carcinoma.

Differences in urease activity in live Helicobacter pylori cultured from patients with gastroduodenal diseases.

AIM: To develop a reliable method for measuring urease activity in live bacteria, and to determine whether there are any differences in urease activity among the Helicobacter pylori strains involved in gastroduodenal disease. DESIGN: The stability of the method was examined in the first phase of the study, and in a second phase the mean urease activity in clinical isolates from different groups of patients was compared. MATERIALS AND METHODS: To assess the stability and reliability of the method, we assessed the relationship between bacterial proliferation and urease activity, the relationship between the number of bacteria and the optical density, and differences in urease activity among bacterial generations. Ten of the 3-day-old colonies in the third generation were suspended in phosphate-buffered saline, and urease activity was measured as 10(5) colony-forming units/ml bacteria. RESULTS: The assay system appeared to be effective, because the urease activity of live bacteria in the logarithmic growth period was constant, the number of bacteria and the optical density showed a linear correlation on a bilogarithmic graph and there was no significant difference in urease activity over three generations. With this method, urease activity varied from 0.192 to 80.42 mIU/10(5) colony-forming units of bacteria/ml. There was no significant difference in the mean urease activity of live bacteria from controls, gastric ulcer patients and duodenal ulcer patients. However, the mean urease activity in bacteria from cancer patients was significantly higher than that of controls or duodenal ulcer patients. CONCLUSIONS: H. pylori strains derived from cancer patients, which have relatively high levels of urease activity, might easily colonize the stomach and lead to much mucosal damage during the long course of H. pylori infection.

Immunogenetic analysis of the human leukocyte antigen DQA1 locus in patients with duodenal ulcer or chronic atrophic gastritis harbouring Helicobacter pylori.

AIM: To examine the human leukocyte antigen (HLA)-DQA1 locus in patients harbouring Helicobacter pylori with chronic atrophic gastritis or duodenal ulcer as part of an investigation into immunogenetic differences in the host. SUBJECTS AND METHODS: We examined 116 patients harbouring H. pylori (55 patients with chronic atrophic gastritis, 61 with duodenal ulcers) and 28 H. pylori-negative healthy controls for HLA-DQA1 genotypes. H. pylori infection was determined by culturing biopsy samples from the gastric body and antrum and by enzyme-linked immunosorbent assay. HLA-DQA1 typing was carried out by the polymerase chain reaction-restriction fragment length polymorphism method. RESULTS: The allele frequency of DQA1*0102 was significantly higher in H. pylori-negative controls (0.250) than in H. pylori-positive duodenal ulcer patients (0.090). In contrast, the allele frequency of DQA1*0301 was significantly lower in H. pylori-negative controls (0.214) than in H. pylori-positive duodenal ulcer patients (0.418). CONCLUSION: These results suggest that there are genetic differences in the HLA-DQA1 locus between H. pylori-positive duodenal ulcer patients and H. pylori-negative healthy controls.

Clonality in Sclerotinia sclerotiorum on Infected Cabbage in Eastern North Carolina.

ABSTRACT Eighty-four isolates of Sclerotinia sclerotiorum from four cabbage production fields in North Carolina and 16 isolates from an experimental cabbage field plot in Louisiana were DNA-fingerprinted and tested for mycelial compatibility. In a comparison with 594 unique DNA fingerprints of S. sclerotiorum from Canadian canola, no fingerprints were shared among Canadian, North Carolina, and Louisiana populations. DNA fingerprints from the North Carolina sample were distinctive from those of the Canadian and Louisiana samples, with significantly more hybridizing fragments in the 7.7- to 18-kilobase range. Forty-one mycelial compatibility groups (MCGs) and 50 unique DNA fingerprints were identified from the North Carolina sample. Three MCGs and three fingerprints were identified from the Louisiana sample. From the North Carolina sample, 32 MCGs were each associated with a unique fingerprint; of these, there were 11 clones (i.e., cases in which two or more isolates belonged to the same MCG and shared the same DNA fingerprint). Six clones sampled from two or more fields represented approximately 29% of the total sample (24 of 84 isolates), with six clones recovered from fields 75 km apart. There were 10 cases in which one MCG was associated with more than one DNA fingerprint and two cases in which one DNA fingerprint was associated with more than one MCG. The small sample from Louisiana was strictly clonal. The North Carolina sample had a clonal component, but deviated from one-to-one association of MCG with DNA fingerprint to an extent consistent with more recombination or transposition than the other two populations sampled.

Early progression stage of malignancy as revealed by immunohistochemical demonstration of DNA instability; I, Human gastric adenomas.

The degree of DNA-instability as revealed by the immunohistochemical staining with monoclonal anti-single-stranded DNA antibody after acid hydrolysis (DNA-instability test) was used as the marker of malignancy. This was applied to human gastric regenerative epithelium in chronic peptic ulcer (5 cases), adenoma (35 cases), and well differentiated tubular adenocarcinoma (5 cases). Proliferative activity was evaluated by proliferating cell nuclear antigen (PCNA) immunohistochemistry, and the quantitative analyses of the mean number and mean area of silver-stained nucleolar organizer regions (AgNORs) per one nucleus were performed for all these cases. All cancers and adenomas were positively stained by the DNA-instability test diffusely, indicating the malignant character of the latter from the view point of DNA-instability, in contrast to the negative stainability of all regenerative epithelium. The percent number of PCNA-positive cells and mean number and mean area of AgNORs tended to be larger in adenoma and cancer than in regenerative epithelium, although the differences were not usually statistically significant. Supporting the malignant character of adenoma, single cell necroses and abnormal mitoses were almost always present in the lesion. In conclusion, all adenoma lesions were regarded as malignant in nature, namely, in-situ carcinoma, existing at an early stage of progression of malignancy.

Importance of acidic mucin secretions by foveolar and mucous neck cells of rat fundic mucosa as the defence mechanisms against HCl as revealed by fasting.

The localization of neutral mucin and acidic mucins in both control and fasted rat gastric fundic mucosa were examined by microscopic and electron microscopic histochemical methods. By Carnoy's fixation, the surface mucous coat of the control rat gastric fundic mucosa was found to be composed of alternating layers of acidic mucins and neutral mucin, indicating the synchronous and cyclic secretions of them. In many gastric pits of the fundic glands, the acidic mucins were found to spring out from the deep foveolar regions like volcanoes. This phenomenon may suggest that the acidic mucins play a fundamental role in protecting the pit cells against HCl during its passage, and the layers of neutral mucin and acidic mucins in the surface coat is the safeguard against the HCl and digestive enzymes in the gastric lumen. In the fasting rat gastric fundic mucosa, the acidity and the amount of the gastric juice were markedly decreased, indicating the suppressed secretions of mucins and HCl. The decreased production of sulfomucin was directly demonstrated by 35SO4-autoradiography. Many mucous neck cells existing in close association with the parietal cells were ballooned due to accumulation of alcian blue (AB)-positive but high iron-diamine (HID)-negative sialomucin, which was not demonstrable in the control. The secretory granules of sialomucin contained in the ballooned mucous neck cells were positively stained ultrastructurally with cacodylate-ferric colloid to stain acid mucopolysaccharides.

Early progression stage of malignancy of human colon border-line adenoma as revealed by immunohistochemical demonstration of increased DNA-instability.

The degree of DNA-instability was used as the marker of malignancy and applied to adenoma (7 benign cases and 17 border-line cases) and cancer (8 carcinoma-in-adenoma cases and 17 invasive cancer cases) of human colon. Proliferative activity by PCNA index and the activity of protein synthesis by AgNORs were also estimated for all cases as the supporting markers of malignancy. In all border-line cases, the following findings were obtained: (1), the degree of DNA-instability as revealed by immunohistochemical staining with anti-single-stranded DNA antiserum after acid hydrolysis was increased in border-line adenoma to the level of invasive overt cancer, indicating its malignancy with marked DNA-instability; (2), reflecting the malignant character, abnormal mitosis and single cell necrosis were usually observed in all border-line adenomas by fluorescent Feulgen staining, indicating the DNA alterations; (3), not only the parenchymal but also the stromal PCNA indices were statistically larger in border-line adenoma than in benign adenoma, indicating the "stromal activation" in malignancy; (4), the volumes of AgNORs were much increased in border-line adenoma in comparison with those in benign adenoma, and these showed further increases with the progression of malignancy to the invasive overt cancer. These findings indicate that border-line adenoma of human colon has already malignant character at the early progression stage, although no apparent epithelial atypia, or destructive invasion, is taking place.

Influence of ammonia solution on gastric mucosa and acetic acid induced ulcer in rats.

Aqueous ammonia in concentrations of 0.02 or 0.1% was continuously administered to rats to study its effect on the gastric mucosa histologically and cell kinetically. Furthermore, acetic acid ulcer, which is a model of chronic gastric ulcer, was experimentally induced in the stomachs of rats to assess the influence of 0.02% ammonia on the course of this ulcer. Male Donryu rats were divided into three groups given 0.02% ammonia, 0.1% ammonia or tap water. On several occasions (1, 3 and 5 days and 1, 4, 8, 12 and 24 weeks from the beginning of the experiment), the gastric mucosa in the fundic gland region and the antrum was examined histologically, and from the viewpoint of cell kinetics. The assessment in the 8th and 24th weeks employed the double labeling technique with bromodeoxyuridine and 3H-thymidine. The assessment on the other occasions used the flash labeling technique with bromodeoxyuridine. Both the 0.02% and 0.1% ammonia treatment groups showed a decrease in PAS-positive mucus and an enhanced cell cycling in the early stage of the experiment. After long periods of treatment, these groups showed a reduction in the gland height, a recovery in PAS-positive mucus and a suppression of cell cycle, suggesting direct toxicity of ammonia on the gastric mucosa. Although glandular atrophy was observed in these animals, infiltration of inflammatory cells was not observed. Thus, the relationship between ammonia and gastritis remained obscure. No ulcer developed in any group. Subsequently, we experimentally induced Ul-IV or Ul-V acetic acid ulcers in the stomachs of rats, according to the method of Okabe et al. (1971, 1972). These rats were divided into two groups given 0.02% ammonia or tap water. In the 4th and 8th weeks of the experiment, the stomachs of these rats were examined histologically and from the viewpoint of cell kinetics. The 0.02% ammonia treatment group showed a significant increase in the ulcer index (long diameter x short diameter; mm2) in the 4th and 8th weeks. This group also showed suppressed cell cycling of the regenerative epithelium and fibroblasts in the ulcer margin, suggesting direct toxicity of ammonia. Thus, healing of peptic ulcer was delayed by continuous administration of 0.02% ammonia.

Antisecretory effect of a PGE2 derivative, enprostil (TA/RS-84135): inhibitory action of amogastrin-stimulated secretion.

A newly developed prostaglandin E2 derivative, enprostil (TA/RS-84135), was administered to five healthy volunteers in the stomach and duodenum through a teflon tube to examine its effect on amogastrin-stimulated gastric acid secretion. The acid output 1-3 h after administration of enprostil (35 micrograms) decreased remarkably as compared with that in the same period after placebo administration. However, no significant difference in antisecretory effect was observed between the two routes of administration. A decrease in pepsin output occurred in parallel with the decrease in acid output. These results suggest that the antisecretory effect of enprostil is at least partially due to systemic absorption and the predominantly topical action seen in animals does not seem to occur in man. No side-effects attributable to enprostil were observed during the test period. In conclusion, enprostil seems to be clinically useful as an antiulcer agent.

A study on the prevention of duodenal ulcer recurrence with pirenzepine.

A double-blind, controlled study with either 50 mg of pirenzepine or a placebo once a day was performed in 128 patients to determine the effect of pirenzepine against relapses of duodenal ulcer, the healing of the subjects' former ulcers having been confirmed endoscopically. The study lasted 12 months at longest; endoscopy was performed at 3,6,9 and 12 month intervals during the study, as well as when subjective symptoms developed. Cumulative non-relapse rates were obtained every 3 months according to the life expectancy table; at 3 and 6 months, they were 85.7% and 73.0% for the pirenzepine group and 67.9% and 49.0% for the placebo group, thus indicating significantly lower relapse rates in favour of the pirenzepine group (p less than 0.05). There was no significant difference between the two groups with regard to the 12-month cumulative non-relapse rate. Comparison of background factors in relapsed and non-relapsed cases suggested that relapses were more frequent among pirenzepine-treated patients who had required a shorter course of treatment or had received H2 antagonists in the treatment of their previous ulcer. Relapses of duodenal ulcer were closely correlated with relapses of subjective symptoms, but there was no significant difference between the pirenzepine group and the placebo group with regard to the frequency of subjective symptoms at the time of ulcer recurrence. Pirenzepine, like H2 antagonists, proved to prevent ulcer relapses temporarily. The relapse rate with the compound appeared to be higher in cases where the former ulcer had been treated with H2 antagonists.

Fulminant Mycoplasma pneumoniae pneumonia with marked elevation of serum soluble interleukin-2 receptor.

We treated a married couple who developed Mycoplasma pneumonia at the same time, and whose clinical courses and serum soluble interleukin-2 receptor (sIL-2R) levels were markedly different. The 30-year-old wife developed acute respiratory failure and her sIL-2R levels were extremely increased. After pulse therapy, her clinical state was improved, with a marked decrease in sIL-2R. In contrast, the 36-year-old husband, also suffered from the pneumonia with a moderate increase of sIL-2R, and recovered without any complications. The difference in their clinical states may be reflected in their serum sIL-2R levels, a marker of T cell activation in vivo.