The Presence of Venous Gas Does Not Affect the Prognosis in Emphysematous Cystitis.

Objective Emphysematous cystitis (EC) has a high mortality rate compared with urinary tract infection without emphysema. However, its prognostic factors have yet to be determined. The presence of venous gas is suspected to be a rare, adverse prognostic factor of EC. However, all four previously reported cases improved. We hypothesized that venous gas is not an adverse prognostic factor of EC and aimed to assess the effect of venous gas on the EC prognosis. Methods Medical records were reviewed retrospectively. Patients The patients diagnosed with EC at Yodogawa Christian Hospital between April 2004 and September 2014 were included. Results Venous gas was present in 15 of 23 patients with EC. There was no significant difference in the background or clinical presentation between patients with or without venous gas. All patients with venous gas survived without invasive measures, whereas 50% of patients without venous gas died. Conclusion There was no marked difference in the mortality rate due to EC between the patients with and without venous gas. Venous gas may be a more common and less worrying finding in EC than assumed. It does not reflect the severity of infection, and air embolisms have not been reported so far. Venous gas may not affect the prognosis. This may be due to the differences in the mechanism of venous gas production. Gas in EC may develop due to glucose fermentation and intravesical pressurization, in contrast to the necrotizing infection seen in other emphysematous infections. This is the first study to assess the effect of venous gas on EC prognosis.

Cocirculation of antigenic variants and the vaccine-type virus during the 2004-2005 influenza B virus epidemics in Japan.

In the 2004-2005 season, there was a large epidemic of the influenza B virus Yamagata group in Kobe, Japan. In hemagglutination inhibition tests, most of the clinical isolates from Kobe showed antigenicities similar to those of previous isolates (the vaccine-type virus). Only a few antigenic variants were isolated around the peak of the epidemic; however, Kobe residents developed antibodies against the variants during the season. The antigenic variants showed a one-point mutation of a nucleotide in the HA1 gene (C440A or G421A), which resulted in the substitution of one amino acid in the 150 loop of the HA molecule (T147N or G141R). The 150 loop is one of four epitopes of the hemagglutinin molecule of the influenza B virus. We established a system to detect one-point differences in the nucleotides of the 150 loop by means of high-resolution melting curve analysis with LCGreen. With this system, the isolates were determined to be the vaccine-type virus, antigenic variants, or a mixture of both. Some isolates were shown to be mixtures although they had been recognized as the vaccine-type virus with the hemagglutination inhibition tests. Thus, the antigenic variants appeared in the early period of the epidemic and were cocirculating with the vaccine-type virus during the epidemic.

Detection of antigenic variants of the influenza B virus by melting curve analysis with LCGreen.

Automated, high-throughput detection methods for single-nucleotide polymorphisms have been applied to the routine genotyping of genetic polymorphisms influencing drug metabolism. Melting curve analysis with LCGreen was introduced recently as one such technique which can be performed rapidly and easily. This technique was used to detect antigenic variants of the influenza B virus. The antigenic variants and vaccine-type strains of the influenza B virus are isolated from clinical specimens of one epidemic season, and they usually differ in one nucleotide in the HA1 gene, corresponding to one amino-acid substitution. By means of melting curve analysis with LCGreen, an antigenic variant clone and a vaccine-type clone were clearly distinguished. In addition, the proportions of the antigenic variants in the mixture-type isolates were estimated. The clinical isolates were detected as the vaccine-type strains, antigenic variants, or a mixture of both. It became clear that humans were infected with a mixture of the vaccine-type strains and the antigenic variants for a certain period after which the viral antigenicities vary. This technique will contribute to the analysis of antigenic shifts in influenza B virus.

Quantification of caveolin isoforms using quantitative real-time RT-PCR, and analysis of promoter CpG methylation of caveolin-1alpha in human T cell leukemia cell lines.

Caveolin-1, an essential structural component of caveolae, functions as a negative regulator for signal transduction and has been suggested to be a candidate tumor suppressor. Lack of caveolin-1 expression has been implicated in the pathogenesis of oncogenic cell transformation and tumorigenesis in many cancers. On the other hand, over-expression has also been associated with tumor progression and metastasis in prostate cancers. Hence, alteration of caveolin-1 expression has been proposed as a clinical marker for diagnosis and prognosis in various cancers. For precise analyses of the caveolin expression in human T cell leukemia cell lines, we measured the mRNA levels of caveolin isoforms, caveolin-1alpha, -1beta, -2, and -3 with real-time RT-PCR using external standards for each isoform. In the panel of human T cell leukemia cell lines tested, four cell lines expressed caveolin-1alpha, -1beta and -2, but not -3, which was consistent with the protein levels. The expression profiles in most cell lines are caveolin-1alpha > caveolin-1beta > caveolin-2. Two cell lines did not express either of the caveolin mRNAs. Methylation analyses for the CpG sites in the promoter region of a positive and a negative cell line did not show a clear correlation with the expression status, suggesting that mechanisms other than CpG methylation are involved in the regulation of caveolin-1alpha expression in human T cell leukemia cell lines.

Differently regulated androgen receptor transcriptional complex in prostate cancer compared with normal prostate.

BACKGROUND: The coregulators of androgen receptors (AR) influence the transcriptional activity of AR. In order to better understand the mechanism of carcinogenesis in the prostate, we investigated the relationship between AR and AR coregulators in the early stage of prostate cancer. METHODS: mRNA was purified from 15 samples of prostate cancer and normal tissue and transcribed into cDNA. We screened eight AR coregulators for different gene expressions in prostate cancer, comparing these with normal tissue by a real-time polymerase chain reaction Syber green method, then quantified each component of the AR transcriptional complex by a real-time PCR hybri-probe method. The extent of gene expression similarity was compared by simple Pearson correlation coefficient analysis between prostate cancer and normal tissue. We applied a z-test to calculate significant differences between r-values. RESULTS: We found that the gene expression level of FHL2 decreased in prostate cancer compared with that of normal tissue and the gene expressions of PSA, AR and SMRT were not significant. The correlation coefficient analysis revealed that strong associations were found in the pairs of AR versus SMRT, AR versus FHL2 and SMRT versus FHL2 in prostate cancer, whereas similarity was found only in the pair of AR versus FHL2 in normal tissue. No association was observed between prostate-specific antigen and other genes. CONCLUSION: These results demonstrate that the AR-AR coregulator relationship is different between prostate cancer and normal tissue, leading to the hypothesis that the AR transcriptional complex is regulated differently between prostate cancer and normal tissue.

Correlation between transcriptional expression of survivin isoforms and clinicopathological findings in human colorectal carcinomas.

Survivin, a novel member of the inhibitor of apoptosis protein (IAP) family, has been markedly overexpressed in most types of human carcinoma, and recognized as a potential target in anticancer therapy. In addition, two splice variants of survivin, survivin-2B and survivin-deltaEx3, have recently been identified. However, expression analysis on its splice variants has not been reported in colorectal carcinomas. Therefore, we investigated the transcription levels of survivin and its splice variants in human colorectal carcinomas, and the correlation between transcript expression levels and pathological findings was analyzed. Tumor tissue samples were provided from 52 cases with colorectal adenocarcinoma resected at the Osaka Medical College from 1995 to 1996. Transcription levels were measured by performing quantitative reverse transcription-polymerase chain reaction (RT-PCR) using primer pairs specific for survivin and either of its splice variants, then normalized by the glyceraldehyde 6-phosphate dehydrogenase. The transcription levels of survivin and its splice variants were significantly higher in the tumor tissue samples, and significantly lower in the normal tissue samples. In addition, approximately 40% of the normal tissue samples did not have any survivin expression. The relative expression level of survivin-2B to survivin (survivin-2B/survivin) was significantly higher in the tumor tissue samples than in the normal ones. In contrast, survivin-deltaEx3/survivin revealed no difference between tumor and normal samples. When Comparing the histological disease stages (stage I and II vs. stage III and IV), there were no significant differences in the expression levels of survivin and its splice variants. The expression level of survivin-2B/survivin for stage III and IV was lower than the one for stage I and II. In addition, a higher level of survivin-2B/survivin significantly correlated with a better prognosis in the present series. The present study demonstrates high expression level of survivin and its splice variants, which is relatively specific in tumor tissue and suggests that they have important roles in the progression of human colorectal carcinomas.

Enhanced detection of malignant glioma xenograft by fluorescein-human serum albumin conjugate.

OBJECT: During the surgical resection of malignant gliomas, it is important to make clear the border zone between the tumor tissue and normal brain tissue. For this purpose, we conjugated fluorescein and human serum albumin (FLS-HSA) and compared its effectiveness against that of fluorescein-sodium (FLS-Na) alone in detecting human glioma xenografts in SCID mice through a fluorescence microscope. METHODS: We made FLS-HSA conjugate using carbodiimide as a linking reagent. SCID mice, with U251MG cells transplanted subcutaneously, were prepared as tumor models. The animals were sacrificed 15, 30, 60, 180, 360, or 720 min after the intravenous administration of either the FLS-HSA conjugate or FLS-Na alone (n = 3). Fluorescence images were taken with a digital camera, and the brightness of the tumor and that of the peripheral tissue in each image were quantified. In the group of tumor-bearing mice that received FLS-Na, the fluorescence of tumor tissue disappeared 60 min after the reagent was administered, and there was no significant difference in brightness between the tumor and peripheral tissue at any time point. On the other hand, injection of FLS-HSA revealed relative tumor-selective brightness and sufficient contrast between the tumor and surrounding tissue 60 and 360 min after administration. CONCLUSION: FLS-HSA has the advantages of specificity and persistence of fluorescence over FLS-Na for the purpose of identifying glioma nodules in xenogenic subcutaneous tumor transplantation models.

Alpha-tocopherol content and alpha-tocopherol transfer protein expression in leukocytes of children with acute leukemia.

Alpha-tocopherol (a form of vitamin E) is a fat-soluble vitamin that can prevent lipid peroxidation of cell membranes. This antioxidant activity of alpha-tocopherol can help to prevent cardiovascular disease, atherosclerosis and cancer. We investigated the alpha-tocopherol level and the expression of alpha-tocopherol transfer protein (alpha-TTP) in the leukocytes of children with leukemia. The plasma and erythrocyte alpha-tocopherol levels did not differ between children with leukemia and the control group. However, lymphocytes from children with leukemia had significantly lower alpha-tocopherol levels than lymphocytes from the controls (58.4 +/- 39.0 ng/mg protein versus 188.9 +/- 133.6, respectively; p < 0.05), despite the higher plasma alpha-tocopherol/cholesterol ratio in the leukemia group (5.83 +/- 1.64 micromol/mmol versus 4.34 +/- 0.96, respectively; p < 0.05). No significant differences in the plasma and leukocyte levels of isoprostanes (the oxidative metabolites of arachidonic acid) were seen between the leukemia patients and controls. The plasma level of acrolein, a marker of oxidative stress, was also similar in the two groups. Investigation of alpha-TTP expression by leukocytes using real-time PCR showed no difference between the two groups. These findings suggest that there may be comparable levels of lipid peroxidation in children with untreated leukemia and controls, despite the reduced alpha-tocopherol level in leukemic leukocytes.

Transcriptional expression of survivin and its splice variants in brain tumors in humans.

OBJECT: Survivin, one of the apoptosis inhibitor proteins, has been detected in most cancers in humans. In addition, two splice variants (survivin-2B and survivin-deltaEx3) have been identified. The authors investigated the transcription levels of survivin messenger (m)RNA and its splice variants in nine tumor cell lines, including gliomas, and in 25 brain tumor samples, by performing quantitative reverse transcription-polymerase chain reaction. The correlation between transcript expression levels and pathological findings were also analyzed. METHODS: Transcription levels were measured using primer pairs specific for survivin and either of its splice variants and were normalized to the glyceraldehyde 6-phosphate dehydrogenase. Among the tumor cell lines tested, glioblastoma cell lines showed the highest levels of survivin expression. Among brain tumor samples studied, survivin was preferentially expressed in malignant brain tumors and gliomas. The relative expression level of survivin-deltaEx3/survivin was significantly higher in malignant than in benign brain tumor samples. Expression patterns were dominant for survivin-deltaEx3 in malignant brain tumors and dominant for survivin-2B in benign ones. A significant linear correlation between survivin mRNA expression and MIB-1 labeling index was demonstrated in all brain tumor samples. CONCLUSIONS: The authors' results indicate that quantifying the levels of survivin and its splice variants is useful for the prediction of the cell biological malignancy of gliomas, independent of their pathological features.

Neutralizing epitopes specific for influenza B virus Yamagata group strains are in the 'loop'.

To study the neutralizing epitopes of influenza B virus Yamagata group strains, two monoclonal antibodies (mAbs) were used to select escape mutants of the virus. mAbs 5H4 and 3A12 were found to react with B/Yamagata group strains in haemagglutination inhibition and neutralization tests; no reactivity with B/Victoria group strains was observed. Most of the mutants reacted poorly to polyclonal ferret antibody against the 1998 isolate. Analysis of the deduced amino acid sequences identified a single amino acid substitution at residue 141 (Gly-->Arg) or 149 (Arg-->Gly) in 5H4-escape mutants and 141 (Gly-->Arg), 147 (Thr-->Ile) or 148 (Ser-->Gly) in 3A12-escape mutants. These residues are situated in close proximity in the 'loop' of the haemagglutinin molecule. These epitopes have been conserved in B/Yamagata group strains for almost 10 years in Japan but amino acid substitutions in the loop have been observed in clinical isolates only since 1999.

a-Tocopherol Content and a-Tocopherol Transfer Protein Expression in Leukocytes of Children with Acute Leukemia.

α-Tocopherol (a form of vitamin E) is a fat-soluble vitamin that can prevent lipid peroxidation of cell membranes. This antioxidant activity of α-tocopherol can help to prevent cardiovascular disease, atherosclerosis and cancer. We investigated the α-tocopherol level and the expression of α-tocopherol transfer protein (α-TTP) in the leukocytes of children with leukemia. The plasma and erythrocyte α-tocopherol levels did not differ between children with leukemia and the control group. However, lymphocytes from children with leukemia had significantly lower α-tocopherol levels than lymphocytes from the controls (58.4±39.0 ng/mg protein versus 188.9±133.6, respectively; p<0.05), despite the higher plasma α-tocopherol/cholesterol ratio in the leukemia group (5.83±1.64 µmol/mmol versus 4.34±0.96, respectively; p<0.05). No significant differences in the plasma and leukocyte levels of isoprostanes (the oxidative metabolites of arachidonic acid) were seen between the leukemia patients and controls. The plasma level of acrolein, a marker of oxidative stress, was also similar in the two groups. Investigation of α-TTP expression by leukocytes using real-time PCR showed no difference between the two groups. These findings suggest that there may be comparable levels of lipid peroxidation in children with untreated leukemia and controls, despite the reduced α-tocopherol level in leukemic leukocytes.

Emergence of an influenza B virus with antigenic change.

Influenza B virus Yamagata group strains, isolated in the 2000 to 2001 influenza epidemic season, reacted poorly to the polyclonal ferret sera prepared against strains isolated earlier. The results of genetic analysis clarified that a point mutation of the nucleotide at position 126 in the HA1 region and the corresponding one-amino-acid substitution altered viral antigenicity.

Antigenic variants with amino acid deletions clarify a neutralizing epitope specific for influenza B virus Victoria group strains.

To study the neutralizing epitopes of influenza B virus Victoria group strains, two monoclonal antibodies (MAbs) were used to select antigenic variants of the virus. MAbs 10B8 and 8E6 were found to react with B/Victoria group strains in three tests, peroxidase-antiperoxidase staining, haemagglutination inhibition and neutralization tests; no reactivity with B/Yamagata group strains was observed. Analysis of the deduced amino acid sequences of 10B8-induced variants identified a single amino acid deletion at residue 165 or 170, as well as a single amino acid substitution at residues 164 (Asp-->Tyr), 165 (Asn-->Ser or Thr) or 203 (Lys-->Thr or Asn). A single amino acid substitution at residue 241 (Pro-->Ser) was observed in 8E6-induced variants. Three-dimensional analysis showed that the epitopes for both MAbs were situated in close proximity to each other. Since B/Yamagata group strains are characterized by amino acid deletions at residues 164-166, the epitope for MAb 10B8 is strictly specific for B/Victoria group strains.