Bone marrow-derived stromal cells are associated with gastric cancer progression.

BACKGROUND: The aim of this study was to clarify the role of bone marrow-derived stromal cells (BM-SCs) expressing CD271 in the development of gastric cancer. METHODS: The effect of human BM-SCs on the proliferation and motility of six gastric cancer cell lines, OCUM-2M, OCUM-2MD3, OCUM-12, KATO-III, NUGC-3, and MKN-74, was examined. CD271 expression levels in BM-SCs were analysed by flow cytometry. We also generated a gastric tumour model by orthotopic inoculation of OCUM-2MLN cells in mice that had received transplantation of bone marrow from the CAG-EGFP mice. The correlation between the clinicopathological features of 279 primary gastric carcinomas and CD271 expression in tumour stroma was examined by immunohistochemistry. RESULTS: Numerous BM-SCs infiltrated the gastric tumour microenvironment; CD271 expression was found in ∼25% of BM-SCs. Conditioned medium from BM-SCs significantly increased the proliferation of gastric cancer cell lines. Furthermore, conditioned medium from gastric cancer cells significantly increased the number of BM-SCs, whereas migration of OCUM-12 and NUGC-3 cells was significantly increased by conditioned medium from BM-SCs. CD271 expression in stromal cells was significantly associated with macroscopic type-4 cancers, diffuse-type tumours, and tumour invasion depth. The overall survival of patients (n=279) with CD271-positive stromal cells was significantly worse compared with that of patients with CD271-negative stromal cells. This is the first report of the significance of BM-SCs in gastric cancer progression. CONCLUSIONS: Bone marrow-derived stromal cells might have an important role in gastric cancer progression, and CD271-positive BM-SCs might be a useful prognostic factor for gastric cancer patients.

Clinicopathological Correlations of Autophagy-related Proteins LC3, Beclin 1 and p62 in Gastric Cancer.

AIM: This study evaluated the clinicopathological significance of autophagy, an intracellular degradation system, in gastric cancer. MATERIALS AND METHODS: The expression levels of three autophagy-related proteins, namely light chain 3 (LC3), Beclin 1 and p62, were analyzed by immunohistochemistry using samples from 510 patients with primary gastric cancer. RESULTS: LC3, Beclin 1, and p62 expression was positive in 79 (15.5%), 126 (24.7%) and 251 (49.2%) out of 510 carcinomas, respectively. Autophagy was defined when samples were positive for at least two out of the three proteins. Autophagy-positive cases were 113 (22.1%) out of the 510. Autophagy determined by LC3, Beclin 1, and p62 significantly correlated with lymph node metastasis, vessel invasion, and hepatic metastasis. A Kaplan-Meier survival curve showed that autophagy was significantly associated with poor survival of patients with gastric cancer, especially for those with disease at stage I. Multivariate analysis indicated that autophagy was an independent prognostic factor. CONCLUSION: Autophagy promotes the progression of gastric cancer at an early clinical stage.

Inverse relationship between the titre of TT virus DNA and the CD4 cell count in patients infected with HIV.

OBJECTIVES: To investigate the prevalence and relative titre of TT virus (TTV) DNA, and to examine the relationship between the extent of TTV viraemia and the immune status among 144 patients with HIV infection; 178 age- and sex-matched healthy individuals were also studied. METHODS: TTV DNA was detected quantitatively by two distinct polymerase chain reaction (PCR) methods [untranslated region (UTR) and N22]. UTR PCR detects all TTV genotypes, and N22 PCR can primarily detect four major TTV genotypes (1-4). RESULTS: Using UTR PCR and N22 PCR, respectively, TTV DNA was detected significantly more frequently in HIV-infected patients than in controls (99 versus 91%, P < 0.001; 56 versus 27%, P < 0.0001), and the relative titre (10N/ml) was significantly higher in HIV-infected patients [4.5 +/- 1.2 (mean +/- SD) versus 3.1 +/- 0.9, P < 0.0001; 2.6 +/- 1.5 versus 1.5 +/- 0.9, P < 0.0001]. Age, sex, co-infection with hepatitis B or C virus, and risk factors for HIV transmission did not appear to be significant factors associated with the titre of TTV viraemia. However, the titre of TTV DNA was significantly higher in HIV-infected patients with AIDS (P < 0.0001), those with low CD4 T cell count (P < 0.0001), or those with high HIV viral loads (P = 0.0047). CONCLUSION: TTV is highly prevalent and high-titred in HIV-infected patients. The TTV viral load may reflect the degree of immune status of these immunocompromised hosts.

Detection of pathogenic Entamoeba histolytica DNA in liver abscess fluid by polymerase chain reaction.

A sensitive method for detection of pathogenic Entamoeba histolytica DNA in drained fluids from liver abscess patients, using the polymerase chain reaction (PCR), has been developed. The PCR employs oligonucleotide primers specific for the gene encoding the 30 kDa molecule of pathogenic E. histolytica. Liver abscess fluids (19 samples), from 14 patients with a presumptive amebic liver abscess, were examined microscopically and by the PCR method. Only two of the 19 samples were positive microscopically, whereas all 19 samples tested positive by PCR. This technique can be used to confirm the diagnosis of amebic liver abscess.

Diagnostic value of culture, histology and PCR for Helicobacter pylori in the remnant stomach after surgery.

BACKGROUND: Diagnosis of Helicobacter pylori infection in the remnant stomach has not been established. AIMS: To investigate the diagnostic value of culture, histology, PCR and serum IgG against H. pylori (ELISA) with and without eradication therapy in the remnant stomach, compared with the unoperated stomach. METHODS: Biopsy samples for bacterial culture and histological diagnosis of H. pylori were taken from the stoma and upper corpus of the remnant stomach and gastric juice was used for PCR assay. RESULTS: Bacterial culture-based diagnosis in the remnant stomach, sensitivity and specificity of culture were 95.1%, 100%; histology 89%, 92.3%; PCR 66%, 89.7%; and ELISA 100%, 50%, respectively, in cases without H. pylori eradication therapy. In assessment of the results of therapy for the remnant stomach, sensitivity and specificity of culture were 100%, 100%; histology 80%, 96.8%; PCR 80%, 91.7%; and ELISA 100%, 0%, respectively. CONCLUSION: Bacterial culture had the highest diagnostic value in the remnant stomach as well as unoperated stomach. Sensitivity by histology and PCR was lower in the remnant stomach than the unoperated stomach, but specificity values were equal. Serum ELISA assay was not suitable for the remnant stomach.

Sex differences in mucosal response to Helicobacter pylori infection in the stomach and variations in interleukin-8, COX-2 and trefoil factor family 1 gene expression.

BACKGROUND: Gastric cancer incidence in men is almost double that in women. We investigated mucosal responses in the stomach against Helicobacter pylori (H. pylori) infections to elucidate the interindividual or sex-related differences, which may in turn be associated with gastric cancer incidence, mucosal changes of stomach as measured by the Sydney System, and interleukin-8, cyclooxygenase-2 and trefoil factor family 1 (TFF1) gene expression. METHODS: An age-, sex-, H. pylori status- and disease-matched case-control study was performed in 574 H. pylori-positive and 225 H. pylori-negative patients selected from 4125 patients with a diagnosis of benign disease of the stomach. Levels of acute and chronic inflammations, atrophy and intestinal metaplasia scored according to the Sydney System were compared by stomach site and by sex. Two biopsy specimens (antral and corpus gastric mucosa) from patients with benign gastric diseases (142 patients; 72 men, 70 women) were analysed for interleukin-8, cyclooxygenase-2 and TFF1 mRNA expression as measured by real-time PCR. RESULTS: Inflammation and activity scores in antrum with H. pylori infection were higher in men, but scores declined according to age. Atrophy and intestinal metaplasia scores in corpus with H. pylori infection appeared more severe in men than in women, especially in older patients. In women, atrophy score increased with increasing age, particularly in postmenopausal H. pylori-negative patients. Interleukin-8 mRNA induction was detected in both antrum and corpus mucosa in H. pylori infection, but sex differences were not found. Response of cyclooxygenase-2 mRNA expression against H. pylori infection in the mucosa was higher in men than women. In H. pylori-negative patients, TFF1 mRNA levels in women were significantly higher than in men, and TFF1 mRNA was significantly lower in positive than negative women. CONCLUSIONS: Sex differences in mucosal responses to H. pylori infection in the stomach may be correlated with sex differences in the incidence of stomach cancer.

A case of AIDS-related hepatic Kaposi's sarcoma.

In Japan, Kaposi's sarcoma (KS) is a very rare neoplasm. However, it does occur as one of the complications in patients with acquired immunodeficiency syndrome (AIDS). AIDS-related KS commonly involves the skin, lymph nodes, and gastrointestinal tract. Hepatic KS is sometimes observed in AIDS patients at autopsy, but it is very rarely diagnosed during life. We report a case of hepatic KS in an AIDS patient, detected by ultrasonography (US), computed tomography (CT), and magnetic resonance imaging (MRI) during life and proven at autopsy. Abdominal US revealed multiple hyperechoic tumors along the portal vein. CT scan showed low density and delayed enhancement by contrast material. These tumors were revealed as a low intensity area on a T1-weighted image of MRI and as a high intensity area on T2-weighted and proton density images. US, CT scan, and MRI revealed characteristic findings of hepatic KS. These procedures are very useful for the diagnosis of hepatic KS. To our knowledge, this is the first report of hepatic KS in Japan.

Studies on bactericidal activities of beta-lactam antibiotics on agar plates: the correlation with the antibacterial activities determined by the conventional methods.

A simplified method to determine minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of beta-lactam antibiotics on agar plates is described. MIC values were determined on agar plates for benzylpenicillin, methicillin and cephalothin using Staphylococcus aureus and Klebsiella pneumoniae. A beta-lactamase solution was then sprayed onto the plates to inactivate the drug(s). After further incubation at 37 degrees C overnight, the minimal concentration at which no test bacteria were visible on the plates was defined as MBC. Both MIC and MBC values decreased with decreased inoculum size. The two values were almost coincidental when high dilutions were used as the inocula. These values were compared with those obtained by the conventional broth dilution method. In this study, MIC as well as MBC values determined by the simplified method were generally smaller than the values determined by the broth dilution technique.

Timed bacteriostatic and bactericidal activities of selected antimicrobial agents against Bacteroides fragilis isolated from clinical specimens.

The activities of selected antimicrobial agents were evaluated for bacteriostatic and bactericidal activities for a large number of clinically obtained strains of Bacteroides fragilis, with special reference to the incubation time of the microbes with the drugs. If the mode of action of a drug is categorized as bactericidal when the ratio of bactericidal concentration/bacteriostatic concentration is low (less than or equal to 4), and as bacteriostatic when high (greater than or equal to 8), during given periods of incubation, then clindamycin, minocycline and chloramphenicol appeared to be bacteriostatic, and cefoxitin, cefmetazole, latamoxef (moxalactam) and metronidazole bactericidal, when the incubation time was brief (6 hours). All these drugs acted bactericidally on most of the test strains, if the time of incubation was prolonged to 24 hours.

Possible beta-lactamase activities detectable in infective clinical specimens.

Biological beta-lactam antibiotic-inactivating activities were detected in bacteriuria and suppurating pleural fluids. Clinical specimens were sterilized with membrane filters and the amounts of ampicillin and/or cephalothin which were being inactivated by 1 ml of each filtrate were determined. In general, filtrates which originally yielded Klebsiella sp. tended to show activity against ampicillin; whereas those yielding Enterobacter sp. and Pseudomonas aeruginosa showed activity against cephalothin.

Comparative bactericidal activities of beta-lactam antibiotics determined in agar and broth media.

Comparative bactericidal activities were determined utilizing a relatively large number of test strains, in both agar and broth media, with special reference to the time of exposure of the bacteria to certain beta-lactam antibiotics. It was apparent that the activities increase with time. The concentrations producing a 99.9% kill with cephalothin for Escherichia coli, Klebsiella sp., and carbenicillin for Pseudomonas aeruginosa were higher in broth than in agar. In contrast, those of benzylpenicillin for alpha-streptococcus (non-enterococcal) were higher in agar than in broth. If the bactericidal concentrations with 3-hour or 6-hour exposure to antibiotics were used as the criterion, these concentrations of carbenicillin for P. aeruginosa, and benzylpenicillin for alpha-streptococcus were, in particular, unusually high compared with the conventionally determined bacteriostatic concentrations (MICs).

Utilization of therapeutic embolization in haemorrhage caused by carcinoma of the tongue.

Selective transcatheter arterial embolization, using Gelfoam, was performed in 2 patients with bleeding from tongue arteries due to carcinoma. Though the patients were in poor general condition, being in the terminal stage of cancer, there were no complications and the bleeding was successfully controlled. This method was effective in controlling haemorrhage from the tongue due to carcinoma of the tongue.

A case of hepatocellular carcinoma in HIV-infected patient.

Persistent human immunodeficiency virus (HIV) infection induces an immuno-suppressive state and therefore malignant tumors are a very common complication. Hepatocellular carcinoma is very rare, however, because it is associated with chronic liver disease by the persistent infection of hepatitis B or C virus (HBV or HCV). We reported a case of HCC with HIV infection who had no evidence of HBV or HCV infection, and that had a rapid growth and active pulmonary metastases. Pathological findings of the resected liver showed moderately differentiated HCC and no chronic liver disease. Despite efforts to find potential HBV integration in tumor and non-tumor tissue, none was observed. To our knowledge, this is the first report of HCC in HIV-infected patient with no evidence of hepatitis virus infection.

Oral administration of fluoroquinolones in the treatment of typhoid fever and paratyphoid fever in Japan.

OBJECTIVE: To study the adverse reactions and therapeutic effects of fluoroquinolones to investigate whether they can be used for the treatment of patients with typhoid fever and paratyphoid fever. METHODS: The adverse reactions and therapeutic effects of fluoroquinolones were studied retrospectively in patients with typhoid fever and paratyphoid fever. PATIENTS: 58 patients (54 Japanese) with typhoid fever, 42 patients (41 Japanese) with paratyphoid fever, and 1 Japanese patient with both typhoid fever and paratyphoid fever, who were admitted in hospitals in Tokyo, Kawasaki, Yokohama, Kyoto, and Osaka from 1995 to 1998 and treated with fluoroquinolones. RESULTS: Almost 80% of the patients were treated with tosufloxacin (TFLX) and the remaining 20 % were treated with norfloxacin, ciprofloxacin, levofloxacin, or sparofloxacin. Side effects (nausea, urticaria, aphthous stomatitis) and elevation of serum amylase were found in 3.6% and 8.3 % of patients treated with TFLX, respectively, but these adverse reactions disappeared in all of these cases either with or without a change in the drug used. No adverse reactions were found in patients treated with the other fluoroquinolones. The clinical and bacteriological effects of these drugs were adequate. CONCLUSION: Though further studies still need to be performed on the fluoroquinolones other than TFLX, we can preliminarily conclude that fluoroquinolones are safe drugs and they can be recommended for the initial therapy of patients with typhoid fever and paratyphoid fever.

Activities of cefoxitin in vitro and in vivo: a study in patients with biliary tract infection undergoing percutaneous transhepatic biliary drainage.

Activities of cefoxitin and the effect on killing of bacteria in bile from six patients undergoing percutaneous transhepatic biliary drainage were analyzed in relation to in vitro activities of the drug, with particular reference to the incubation time. The bacteriostatic and bactericidal activities of cefoxitin with short term (6 hr) incubation correlated well with the duration of the effective cefoxitin concentration and the decrease of viable bacteria in bile juice from the patients. These timed in vitro activities can be used as well as the conventional MIC and MBC with the prolonged incubation of 24 hr, in the deductive analysis of bacterial response in vivo.

[Study on bacterial infection by the investigation of antimicrobial susceptibility of various clinical isolates. III. Comparison of drug susceptibility between the isolates from the materials of out- and in-patients, and among the isolates from the materials of different hospital units].

Antimicrobial agents susceptibility of 42, 940 strains of clinical isolates (from 1979 to 1986) were investigated and the data were analyzed on the basis of the sources of isolates; materials from in- and out-patients or from the different hospital units. Bacteria studied were limited to the species of which isolates were 100 or more during 1979 to 1986. The following results were obtained. i) When the antibiotic susceptibility of the isolates from the out-patient-materials were compared with that from the in-patient-materials, the following 13 bacterial species isolated from the former source were found to be more susceptible to antibiotics than that of the latter. These were S. aureus, S. epidermidis, E. coli, Citrobacter sp., Klebsiella sp., Serratia sp., P. vulgaris, P. rettgeri, M. morganii, P. aeruginosa, P. putida, S. sonnei and Salmonella sp.. ii) The frequency of isolation and their antibiotic susceptibility of S. aureus, Enterobacter sp. and P. aeruginosa from both the in- and out-patients were comparable through 1979. However, S. aureus isolated from the in-patient-materials tended to show increased antibiotic resistance since 1983. This is probably due to the frequent use of the 3rd generation cephalosporins. iii) Comparison of the antibiotic susceptibility of the isolates from the different hospital units showed that the resistant strains were more frequently isolated in the materials from the urology unit and the susceptible strains were more frequently isolated in the materials from the infectious diseases unit. iv) Antibiotic resistant S. aureus and P. aeruginosa increased abruptly since 1983 in the materials from the surgery and urology unit, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

[Fundamental studies on serological diagnosis of amoebiasis. 2. Application of antigen of Entamoeba histolytica for complement fixation test to counterimmunoelectrophoresis; and clinical relevance of serological tests].

Antigen of Entamoeba histolytica for complement fixation (CF) test was applied to counterimmunoelectrophoresis (CIE), and the result was compared with that of CF test and enzyme-linked immunosorbent assay (ELISA) in regard to sensitivity and specificity. Besides, we studied antibody titers to E. histolytica in sera from acute and convalescent patients, and from patients with amoebic colitis and amoebic liver abscess. Sera used were from patients with amoebic colitis, amoebic liver abscess and uninfected control subjects. The CIE was less sensitive than the CF test and ELISA. Most CIE-negative samples had low CF and ELISA titers, but a few samples had higher CF and ELISA titers. On the other hand, all uninfected controls were negative by CIE and ELISA, and 98% were negative by the CF test. The level of antibodies tended to decrease with time after clinical recovery, but CF test and ELISA results remained positive in all patients 6 months after clinical cure. CIE converted negative in only one patient after 6 months. The level of antibodies of CF test and ELISA was significantly higher in amoebic liver abscess than amoebic colitis. These results suggested that all these three methods were so specific, and antibody titers by the CF test and ELISA were well correlated to clinical manifestations. The CIE with CF-antigen was less sensitive as compared with CF test or ELISA.

[Clinical research on patients with typhoid and paratyphoid fever (1984-1987). Research Group for Infectious Enteric Diseases, Japan].

Patients with typhoid or paratyphoid fever, admitted to 14 hospitals for infectious diseases during 1984-1987, were epidemically and clinically studied. Of the total number of 183 typhoid, 49 paratyphoid fever patients, those infected overseas was 44.3% and 71.4% respectively, giving an overall annual decrease, yet marking an increased ratio of overseas infection. Patients aged 20s-30s and males were dominant. One hundred and seventy six cases (96.2%) of typhoid and all the paratyphoid fever cases were bacteriologically diagnosed. The period from the onset to the diagnosis was around 14 days in most cases, but beyond 29 days in over 10% of the cases. We would like to emphasize that enteric fever, focusing on high fever, bradycardia, roseola, hepatosplenomegaly, leukopenia, elevated serum-GOT GPT and LDH, can be easily diagnosed by blood/stool culture before beginning chemotherapy. Intestinal bleeding was recognized in 24 cases (13.1%) of typhoid and 4 (8.2%) of paratyphoid fever, intestinal perforation in 2 (1.1%) and death in 1 (0.5%) of typhoid fever. CP was most commonly used in chemotherapy. Bacteriological relapse was recognized in 7/127 cases (5.5%) of typhoid, 6/48 (13.0%) of paratyphoid fever those followed beyond 3 weeks, though eradication was attained by retreatment. One strain of S. typhi resistant to CP.ABPC.KM.SM was isolated in 1986 from a patient infected overseas. New quinolones seem reliable in our preliminary studies.

[Varicella in adulthood: clinical features, severity scores, source of infection and complications].

Varicella has been thought to be one of the representative infectious disease in childhood, but recently we are under the impression that adults contracting varicella are increasing in number. On the other hand, they say that varicella generally causes a serious illness in adult patients. So we investigated signs and symptoms of varicella, source of infection, occupations of adult patients, except those who were immunologically compromised, by means of medical records, to know the characteristics of varicella in adulthood. According to the varicella severity score proposed by Nagai et al., varicella in the hospitalized adult patient was found to be much severer than that in children. The most remarkable symptoms, were high fever and sore throat, and these were the main reason of hospitalization in most of our patients. Although severity scores were very high in admitted adult patients with varicella, their clinical courses were not serious, and most of them recovered with only supportive therapy. These patients rarely suffered from complications, like pneumonia. If adult patients with varicella hospitalized in the early stage and received supportive care, they could recover without any complications. In most cases of adult varicella the source of infection was unknown. In the case of married persons, however, many of them were infected through their child. When adults contract varicella, not only the patients themselves suffer from high fever and sore throat, but also they act as the source of infection, if they are medical care workers. Furthermore, in public, the contraction of varicella results a socioeconomic loss from suspension of business caused by the illness. Prophylaxis with varicella vaccine, therefore, should be considered, when there are people who have never contracted varicella, whether or not they are medical staff.

[In vitro drug susceptibility test of Plasmodium falciparum using a portable thermostat and CO2 gas generator].

We have recently developed a method of in vitro cultivation of P. falciparum using a portable incubator and AnaeroPack.CO2 (Onda et al.), we applied semi-microtechnique drug susceptibility tests to the culture method to evaluate the system using several P. falciparum strains or isolates of different susceptibilities to chloroquine (SGE-1, FCR-3, K-1, Patient 1 and 2). The new method gave comparable results to those shown by the standard test employing a modular incubator chamber with standard gas composition of 5% O2, 5% CO2 and 90% N2. Many useful data on the epidemiology of drug resistant malaria such as the emergence of multi-drug resistant isolates could be collected by applying this new method to the field survey.