Down-regulation of the two-component system and cell-wall biosynthesis-related genes was associated with the reversion to daptomycin susceptibility in daptomycin non-susceptible methicillin-resistant Staphylococcus aureus.

Daptomycin (DAP) is widely used in the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infection. The emergence of DAP non-susceptible MRSA strains during therapy is a major concern in clinical settings. Recent studies revealed that MRSA spontaneously reverts to a subsequent methicillin-susceptible S. aureus (MSSA) strain. However, it is not clear whether DAP non-susceptible MRSA has the ability to revert to a susceptible strain. We obtained an MRSA strain pair, DAP non-susceptible strain and subsequent DAP susceptible strain, from a patient. To understand the underlying mechanism by which DAP non-susceptible MRSA reverts to a susceptible strain, we performed genetic and phenotypic analysis in the strain pair. Although whole-genome analysis revealed four missense mutations, including L826F in mprF, in both strains, the net cell-surface charge was similar between the DAP non-susceptible and susceptible strains. However, the thickness of the cell wall was higher in the DAP non-susceptible strain, which was decreased to the same level as the control after reversion to the DAP susceptible strain. Moreover, the non-susceptible strain showed higher mRNA expression of the two-component system (TCS), such as VraSR, yycG and GraS, with the up-regulated transcription levels of cell-wall biosynthesis-related genes. The expression levels of those genes were decreased after reversion to the susceptible strain. These results indicated that DAP non-susceptibility due to up-regulation of the TCS and cell-wall biosynthesis-related genes may be reversible by the discontinuation of DAP, leading to reversion to the DAP susceptible phenotype.

[Efficacy of the urine antibody test for detection of Helicobacter pylori: comparison with serum antibody tests].

A urine-based enzyme-linked immunosorbent assay kit (URINELISA) for detection of the antibody for H. pylori has recently been developed. We evaluated the diagnostic capability of the URINELISA test in comparison with two serum IgG antibody kits (HM-CAP and Helico G) for H. pylori infection. The subjects of this study were 173 patients. H. pylori was detected by means of culture, immunohistochemical staining and rapid urease test of endoscopically obtained biopsy specimens. A positive diagnosis of H. pylori was when at least one of three tests was positive and a negative diagnosis when all three were diagnosed as H. pylori positive and 23 as negative. We also examined the diagnostic potential of the antibodies in urine and in serum by using the results of the bioptic methods as the gold standard. The sensitivity of both URINELISA and HM-CAP was 93.3% and that of Helico G 94.7%, while their respective specificities were 47.8%, 65.2% and 52.2%. The URINELISA kit thus showed high sensitivity but relatively low specificity. The latter was due to sampling errors in the bioptic procedure because the subjects of this study included many elderly patients with atrophic gastritis. The level of the antibody in urine decreased markedly after 2 months of eradication therapy. We conclude that the URINELISA kit is as effective as serum antibody kits for the diagnosis of H. pylori infection.

[Changes in serum anti-Helicobacter pylori IgG antibody, pepsinogen I, and pepsinogen II after eradication therapy of Helicobacter pylori].

To investigate the changes in serum anti-Helicobacter pylori IgG antibody (HP Ab), pepsinogen I (PG I), pepsinogen II (PG II), and pepsinogen I/II ratio (PG I/II) after eradication therapy of Helicobacter pylori (HP), we studied 78 patients with HP-positive peptic diseases. They received combination therapy (proton pump inhibitor + amoxicillin: n = 17, proton pump inhibitor + amoxicillin + clarithromycin: n = 61). In the 68 patients in whom HP was eradicated, HP Ab, PG I, and PG II decreased and PG I/II increased significantly after eradication. Especially, the decrease in PG II and the increase in PG I/II were rapid and remarkable, found 2 months after the beginning of eradication therapy, and then continued. On the other hand, in the patients in whom HP was not eradicated, HP Ab and PG I/II did not change significantly, while PG I and PG II temporarily increased at the end of administration of proton pump inhibitor. In conclusion, it seems that the measurement of PG II and PG I/II is useful for the early detection of HP eradication.

Long-term follow-up of mitomycin C nephropathy.

Two cases of mitomycin C nephropathy, which occurred after postoperative chemotherapy for advanced gastric cancer, were followed up for 6 (case 1) and 10 years (case 2). Hemolytic uremic syndrome developed 68 days (case 1) and 160 days (case 2) after the last administration of MMC with a total dose of 60 mg (case 1) and 40 mg (case 2). Serum creatinine levels were normalized in case 1 and they remained at about 2 mg/dl in case 2. Hyporeninemic hypoaldosteronism was transiently seen in case 2. These data suggest that recovery from the acute phase of hemolytic uremic syndrome leads to a good long-term prognosis in MMC nephropathy.

Measurement of sialylated stage-specific embryonic antigen-1 in pure pancreatic juice for the diagnosis of pancreatic cancer.

The diagnostic significance of measuring sialylated stage-specific embryonic antigen-1 (SLX) in pure pancreatic juice was evaluated in 20 patients with pancreatic cancer, 43 with chronic pancreatitis, 13 with cholecystolithiasis, and 15 control individuals. Four fractions of pure pancreatic juice were collected sequentially from the pancreatic duct by endoscopic cannulation. The SLX levels in all four fractions of pure pancreatic juice were significantly higher in patients with pancreatic cancer than in controls. On the other hand, patients with chronic pancreatitis or cholecystolithiasis did not have SLX levels that significantly differed from those of controls in any fraction. When the cut-off value was set as the mean concentration +2 times the standard deviation of the control values, the positive rates of SLX in the first fraction (washout phase) and the third fraction (secretory phase) of pure pancreatic juice from pancreatic cancer were 55% (11/20) and 40% (8/20), respectively. Although the false positive rates in the first fraction were high in chronic pancreatitis (30%) and cholecystolithiasis (31%), such high SLX levels in the third fraction were found only in one (2%) patient with chronic pancreatitis and in one (8%) with cholecystolithiasis. The specificities of the test for pancreatic cancer in the first fraction and the third fraction were 70% (39/56) and 96% (54/56), respectively. These results indicate that the measurement of SLX in the third fraction of pure pancreatic juice is useful as a specific marker for pancreatic cancer.

[Availability of determination of serum anti-Helicobacter pylori IgG antibody in diagnosis of Helicobacter pylori infection].

We studied on clinical availability of determination of serum anti-Helicobacter pylori (HP) IgG antibody in diagnosis of HP infection compared with bacterial culture. The population of this study consisted of patients who underwent endoscopic examination in our hospital. Detection and quantification of HP were made by the culture of biopsy specimens taken from the antrum and the body of the stomach. And, simultaneously, serum anti-HP antibody (HP-Ab) was measured by ELISA. The levels of HP-Ab in culture positive patients, 40.6 +/- 33.7U, are significantly higher than that of culture negative patients, 10.6 +/- 9.1U. Sensitivity and specificity in diagnosis of HP infection using determination of serum HP-Ab were 96.2% and 60.0% respectively. Significant correlation was found between the levels of serum HP-Ab and the amounts of HP bacilli in the biopsy specimen taken from body of the stomach.

Diagnostic significance of cancer-associated carbohydrate antigen (CA19-9) concentrations in pancreatic juice: analysis in pure pancreatic juice collected by endoscopic aspiration and immunohistochemical study in chronic pancreatitis.

This study evaluated the diagnostic significance of concentrations of the cancer-associated carbohydrate antigen CA19-9 in pure pancreatic juice (PPJ) collected by endoscopic cannulation. We also attempted to elucidate the features and source of the increased CA19-9 concentration found in the pancreatic juice of patients with chronic pancreatitis (CP) by means of immunohistochemical staining. The mean output as well as the mean concentration of CA19-9 in each of the four fractions collected was highest in patients with pancreatic cancer (PC) and also was elevated significantly in patients with CP compared with controls. However, CA19-9 concentrations were not elevated in patients with cholecystolithiasis. When the cutoff value was set as the mean concentration + 2SD of the controls, significantly elevated concentrations of CA19-9 were found in the third fraction (secretory phase) in 90% of the patients with PC and 66% of the patients with CP. Immunohistochemical staining revealed that CA19-9 was expressed more widely in the ductal cells of CP tissues than in those of normal pancreatic (NP) tissues, with CP tissue showing more CA19-9-positive ductal cells per area than NP tissues. In NP tissue, CA19-9 was localized to the apical surface and supranuclear regions (apical type) in all the ductal cells stained by the antigen, while approximately 50% of cases with CP exhibited a cytoplasmic pattern showing a loss of polarity of the antigen expression. Moreover, this cellular localization pattern was more pronounced in the small ducts that had proliferated and aggregated following the destruction of lobules in CP.(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of various sialylated carbohydrate antigens in malignant and nonmalignant pancreatic tissues.

The expression of six sialylated carbohydrate antigens (CA19-9, CA-50, SLEX, SLX, DU-PAN-2, ST-439) was examined in malignant and nonmalignant pancreatic tissues using an immunohistochemical method to elucidate the characteristics of these carbohydrate antigens as tumor markers. All carbohydrate antigens except for sialyl SSEA-1 (SLX, 52.4%) were expressed in more than 80% of the pancreatic cancer. CA19-9 and CA-50, belonging to type I blood group antigens, and DU-PAN-2 and ST-439 were localized predominantly in the cytoplasm of cancer cells, while sialyl Lex (SLEX) and SLX, belonging to type II blood group antigens, were stained mainly on the apical membranes of malignant glands. Although type I antigens were expressed in most nonmalignant pancreatic tissues, the type II antigens and ST-439 were absent in almost all of the normal tissues and faintly expressed in few chronic pancreatitis tissues, suggesting the high tumor specificity of these antigens. Each antigen was expressed on the apical surface of ducts in normal pancreas. However, in about 30% of chronic pancreatitis cases, type I antigens and DU-PAN-2 were observed in the cytoplasm of ductal cells. All patients showing stromal stain, possibly caused by loss of antigen polar expression and shedding into the surrounding stroma adjacent to malignant glands, revealed high levels of serum antigen. This finding suggests that the stromal appearance of antigens is a significant factor in the elevation of serum antigen levels.

Determination of mono-O-acetylated N-acetylneuraminic acids in human and rat sera by fluorometric high-performance liquid chromatography.

A simple, rapid, and highly sensitive fluorometric high-performance liquid chromatographic method for the determination of N-acetylneuraminic acid and its mono-O-acetyl derivatives in human and rat sera is described. The neuraminic acids, released by hydrolysis of serum in 2 M acetic acid, are converted with 1,2-diamino-4,5-methylenedioxybenzene, a fluorogenic reagent for alpha-keto acids, to highly fluorescent derivatives without the occurrence of O-acetyl migration and de-O-acetylation. The derivatives are separated isocratically within 25 min by reversed-phase chromatography using a TSK gel ODS-120T column. The limits of detection are 57-192 fmol in a 10-microliters injection volume at a signal-to-noise ratio of 3. This sensitivity permits precise determination of the neuraminic acids in 5 microliters of human and rat sera.

[Clinical evaluation of serum sialyl SSEA-1 (SLX) in diagnosis of cancers].

In order to evaluate the usefulness of sialyl SSEA-1 (SLX) as a tumor marker of digestive cancers, serum levels of the antigen were determined in 334 patients with malignancies and 196 patients with benign diseases. The results indicated that positivity of the antigen in sera from malignant patients was highest in pancreatic cancer (58%) and biliary tract cancer (56%). False positive incidence of SLX in sera from benign diseases was as low as 6%, revealing low false positivity. Comparison with other tumor markers such as CA19-9, CA-50, CEA and ST-439 showed that positivity of SLX was as high as that of CEA, whereas it was lower than that of CA19-9 or CA-50. On the other hand, false positivity of SLX as well as ST-439 was lowest, and accuracy of SLX was no less high than that of CA19-9 or CA-50. In sera of pancreatic and biliary tract cancer, positive incidences of CA19-9, CEA and ST-439 were 80%, 64% and 53%, respectively, and the diagnostic efficiency increased by combined assay of SLX with CA19-9 (88%), CEA (81%) and ST-439 (71%). SLX appears to be no less useful than the other recently developed carbohydrate antigens or CEA as serum tumor marker for pancreatico-biliary cancer.

[Analysis of complications following double contrast arthrotomography of the temporomandibular joint].

Temporomandibular joint (TMJ) arthrography is a valuable diagnostic method to evaluate the pathology of soft tissue components of the TMJ. However, arthrography is more or less invasive technique. The aim of this study was to investigate the sort and content of injury following TMJ arthrography. The symptoms before and after the arthrography were compared. In 90 joints that underwent TMJ arthrography, complications such as pain and/or trismus were encountered in 39 joints within one month. Twenty six patients complained of the change of pain conditions including the discomfort. The most common one was the pain on mouth opening. In every joint the pain disappeared within one month without any treatment. There was a high frequency of exacerbated pain in patients with some pain before the arthrography. The change of the degree of maximum opening was recognized in 11 patients. The duration of that condition was somewhat longer than that of pain conditions. In 5 joints with late clicking, 3 developed closed lock. Other complications were swelling, disability of mastication, eczema, hearing impairment and facial paresthesia. All the symptoms disappeared within 1 week without any treatment. From these results it was suggested that TMJ arthrography has low possibility of severe damage to the TMJ although there is some possibility of injury to the bone and soft tissue by arthrographic procedures.

[Cancer-associated carbohydrate antigens available for serum diagnosis].

Many of the recently developed tumor antigens detected by monoclonal antibodies are sugar chains and frequently associated with blood group substance. By immunohistological studies, we evaluated mainly the clinical usefulness and significance of the serum assay of CA-50 classified as a type 1 sugar chain, sialyl SSEA-1 classified as a type 2 sugar chain, and ST-439 with an undetermined structure, as well as their clinicopathological significance. In addition, the value of measurement of CA 19-9 and ST-439 in pancreatic juice was studied. The incidence of serum CA-50 was highest in pancreatic cancer (86%) and biliary tract cancer (66%), but relatively low in benign diseases. Moreover, the positivity and specificity of CA-50 as a tumor marker were no less useful than those of CA 19-9. However, comparison of serum levels of CA-50 with those of CA 19-9 in the same samples revealed a highly positive correlation in malignant diseases. Thus, improvement of the diagnostic rate through a combination assay of CA-50 and CA 19-9 seems unlikely. Both serodiagnostic and immunohistological studies showed that sialyl SSEA-1 and ST-439 were highly specific for tumor, whereas their appearances in serum or tumor were lower than CA 19-9 or CA-50 carrying the type 1 sugar chain. In addition, an appreciable number of sialyl SSEA-1 or ST-439 positive patients were found among those negative for CA 19-9 or CA-50. These results indicate that assay of sialyl SSEA-1 or ST-439 improves the diagnostic rate in combination with CA 19-9 or CA-50. Although the concentration of CA 19-9 in pancreatic juice from pancreatic cancer patients was highest, it was also significantly higher in patients with chronic pancreatitis than in controls. Furthermore, the overlap between the values in patients with cancer and those with chronic pancreatitis was great. On the other hand, the concentration of ST-439 in pancreatic juice was significantly higher only in patients with pancreatic cancer. These results indicate that the measurement of ST-439 in pancreatic juice is more useful as a tumor marker than measurement of CA 19-9, and that moreover, the assay of CA 19-9 in pancreatic juice could be used as a sensitive marker for nonspecific pancreatic injury.