CSC MARKERS IN OSCC AND THEIR ASSOCIATION WITH CLINICAL FACTORS

Cancer stem cells (CSCs) exhibit self-replication, self-differentiation, drug resistance and immune evasion activities. In recent years CSCs have become increasingly important for the treatment of malignant tumors. CSCs express specific markers, including cluster of differentiation (CD)44, CD44 variant 9 (CD44v9), ATP-binding cassette sub-family G member 2 (ABCG2), CD24, B lymphoma Mo-MLV insertion region 1 homolog (BMI-1) and aldehyde dehydrogenase 1 (ALDH1). However, the prognostic value of their expression in patients with oral squamous cell carcinoma (OSCC) are not well known. The present study evaluated these markers in stage I and II patients with OSCC and examined the association between T classification, histological differentiation, classification of invasion mode, lymph node metastasis and disease-free survival rate. Tissue specimens were obtained from 70 patients with stage I or II OSCC following either surgery or biopsy. Immunohistochemistry was performed and positive staining was defiend as 10% positive cells. CD44 and CD44v9 expressions were strongly detected in all OSCC tissues compared with normal epithelial cells. A total of 22 (31.4%) cases expressed ABCG2 and there was a significant association between ABCG2 expression and invasion. A total of 41 cases (59.0%) expressed CD24 and there was a significant association between CD24 expression and invasion. A total of 33 cases (47.1%) expressed BMI-1 and there was a significant association between BMI-1 expression and the disease-free survival rate. A total of 18 cases (25.7%) expressed ALDH1. Although there was no association between ALDH1 expression and T classification, there were significant associations between ALDH1 expression and histological differentiation, invasion mode, metastasis and the disease-free survival rate. Multivariate analysis revealed that ALDH1 expression was the only prognostic factor for disease-free survival rate. The results of the present study suggest that the positivity of ALDH1 detected in patients with OSCC correlates with the number of cells undergoing epithelial mesenchymal transition and metastasis. These findings indicated that the expression of ALDH1 may be an effective prognostic marker indicating the survival of patients with stage I and II OSCC

Isolation and characterization of mutations affecting expression of the Δ9- fatty acid desaturase gene, OLE1, in Saccharomyces cerevisiae

AbstractExpression of the Δ9- fatty acid desaturase gene, OLE1, of Saccharomyces cerevisiae is negatively regulated transcriptionally and post-transcriptionally by unsaturated fatty acids. In order to isolate mutants exhibiting irregulation of OLE1 expression, we constructed an OLE1p–PHO5 fusion gene as a reporter consisting of the PHO5 gene encoding repressible acid phosphatase (rAPase) under the control of the OLE1 promoter (OLE1p). By EMS mutagenesis, we isolated three classes of mutants, pfo1, pfo2 and pfo3 (positive regulatory factor for OLE1) mutants, which show decreased rAPase activity under derepression conditions (absence of oleic acid). Analysis of the transcription of OLE1 in these pfo mutants revealed that pfo1 and pfo3 mutants have a defect in the regulation of OLE1 expression at the transcriptional level while pfo2 mutants were suggested to have a mutation affecting OLE1 expression at a post-transcriptional step. In addition, four other classes of mutants, nfo1, nfo2, nfo3 and nfo4 (negative factor for OLE1) mutants that have mutations causing strong expression of the OLE1p–PHO5 fusion gene under repression conditions (presence of oleic acid), were isolated. Results of Northern analysis of OLE1 as well as OLE1p-PHO5 transcripts in nfo mutants suggested that these mutations occurred in genes encoding global repressors. We also demonstrated that TUP1 and SSN6 gene products are required for full repression of OLE1 gene expression, by showing that either tup1 or ssn6 mutations greatly increase the level of the OLE1 transcript

Two-stage sinus floor augmentation using carbonate apatite

Purpose: The purpose of this study was to elucidate the efficacy and safety of carbonate apatite (CO3Ap) granules in 2-stage sinus floor augmentation through the radiographic and histomorphometric assessment of bone biopsy specimens. Methods: Two-stage sinus floor augmentation was performed on 13 patients with a total of 17 implants. Radiographic assessment using panoramic radiographs was performed immediately after augmentation and was also performed 2 additional times, at 7±2 months and 18±2 months post-augmentation, respectively. Bone biopsy specimens taken from planned implant placement sites underwent micro-computed tomography, after which histological sections were prepared. Results: Postoperative healing of the sinus floor augmentation was uneventful in all cases. The mean preoperative residual bone height was 3.5±1.3 mm, and this was increased to 13.3±1.7 mm by augmentation with the CO3Ap granules. The mean height of the augmented site had decreased to 10.7±1.9 mm by 7±2 months after augmentation; however, implants with lengths in the range of 6.5 to 11.5 mm could still be placed. The mean height of the augmented site had decreased to 9.6±1.4 mm by 18±2 months post-augmentation. No implant failure or complications were observed. Few inflammatory cells or foreign body giant cells were observed in the bone biopsy specimens. Although there were individual differences in the amount of new bone detected, new bone was observed to be in direct contact with the CO3Ap granules in all cases, without an intermediate layer of fibrous tissue. The amounts of bone and residual CO3Ap were 33.8%±15.1% and 15.3%±11.9%, respectively. Conclusions: In this first demonstration, low-crystalline CO3Ap granules showed excellent biocompatibility, and bone biopsy showed them to be replaced with bone in humans. CO3Ap granules are a useful and safe bone substitute for two-stage sinus floor augmentation

Characterization of Nocardia asteroides Isolates from Different Ecological Habitats on the Basis of Repetitive Extragenic Palindromic-PCR Fingerprinting

Thirteen isolates of Nocardia asteroides from both soils and aquatic samples (lake and moat sediments, as well as scum from activated sludge), together with a type strain and two known clinical isolates of this species, were characterized by repetitive extragenic palindromic-PCR fingerprinting with the BOX-A1R primer. The resulting DNA fingerprint patterns proved to be strain specific, and cluster analysis distinguished the soil isolates, the aquatic isolates, and the known strains as being in separate groups

Identification of CDC25-P1306L, a novel mutant allele of CDC25, conferring tolerance to multiple stresses associated with food production on Saccharomyces cerevisiae

Mutations conferring tolerance to diverse stresses (i.e. multistress tolerance) on budding yeast Saccharomyces cerevisiae are useful for industrial yeast strains such as baker’s and wine yeast. However, little is known about the mutations conferring multistress tolerance. Previously, we developed a simple method for isolating multistress-tolerant semidominant mutants of S. cerevisiae by one-step selection under lethal hydrogen peroxide stress, which we named the LCH method. In this study, we applied a combination of genetics and next-generation sequencing (NGS) technology to identify the causal mutation for multistress tolerance of the mutant isolated using the LCH method. The haploid mutant strain was crossed with the wild-type strain and the resulting diploids were sporulated. The 20 haploid progeny strains showing multistress tolerance were mixed and subjected to DNA extraction for NGS. CDC25-P1306L, a novel mutant allele of CDC25 encoding a Ras guanine nucleotide exchange factor, was detected 86 times; however, a wild-type CDC25 allele was not detected in the NGS data from the mixture of the multistress-tolerant progeny strains, suggesting that all of the progeny strains showing multistress tolerance have a CDC25-P1306L allele instead of a wild-type CDC25 allele. Substitution of CDC25 in the wild-type strain with CDC25-P1306L rendered the strain tolerant to ethanol, heat shock, freeze-thaw, chronological aging and high concentrations of glucose. These results indicate that CDC25-P1306L is a multistress-tolerant mutation and is promising for breeding multistress-tolerant S. cerevisiae strains for food production

Glucose repression of FLO11 gene expression regulates pellicle formation by a wild pellicle-forming yeast strain isolated from contaminated wine

Some wild strains of Saccharomyces cerevisiae form a pellicle on the surface of contaminated wines during the post-fermentation period of wine making. In this study, we found that both pellicle formation and FLO11 expression by a wild pellicle-forming strain of S. cerevisiae isolated from contaminated wine were repressed in a glucose-containing medium. Substitution of the promoter region of FLO11 in the cells with a constitutive promoter caused derepression of pellicle formation in the glucose-containing medium and wine. These findings indicate that glucose repression of the expression of the FLO11 gene, but not the glucose repression of the expression of other glucose-repressed genes, is responsible for the glucose-dependent regulation of pellicle formation by the wild pellicle-forming yeast strain. Furthermore, we found that the wild pellicle-forming strain had the same deletion of the 111-bp repression sequence in the FLO11 promoter as the flor strains commercially used for making sherry-like wines. Based on the results obtained in this study, a new method that would prevent deleterious pellicle formation in wineries is discussed

Traditional microbial control methods used in sake brewing effectively suppress predominant bacteria emerging during production of rice koji

AbstractRice koji is a raw material used in the production of sake; however, details regarding the microbial flora and their dynamics in rice koji during the production process are poorly understood. Clarifying these issues can contribute to proposing a method and evaluation that will improve the quality of rice koji and sake. The aim of this study was to determine the microflora in rice koji and the effectiveness of the traditional microbial control techniques used in the sake production process. We analyzed the diversity and changes in bacterial flora during rice koji production by amplicon sequencing of the 16S rRNA gene. The predominant taxon in all rice koji samples was family Staphylococcaceae. The microbial population and the changes in its distribution for five consecutive stages in rice koji production were examined by direct colony counting. Bacteria counts in all samples were below the limit of detection initially, then increased rapidly toward the final stage. The predominant bacterial colonies from all samples were yellow and were identified as Staphylococcus gallinarum through 16S rRNA gene sequence similarity. The S. gallinarum isolates exhibited faster growth in pregelatinized rice medium. Interestingly, the growth of S. gallinarum isolates was suppressed by low temperature (12 °C), ethanol concentration (≥6%) and the addition of lactic acid, which are traditional microbial control methods used during sake fermentation. Therefore, proper control of the traditional sake production process can effectively inhibit the growth of undesirable bacteria such as S. gallinarum that emerge during the production of rice koji

A case of gallbladder torsion diagnosed by abdominal CT prior to surgery

A 77-year-old female with a chief complaint of epigastralgia was diagnosed with torsion of the gallbladder by abdominal CT prior to surgery and successfully treated by an emergency operation. Only 16 cases of gallbladder torsion including the present case have been diagnosed prior to surgery in Japan. Almost all of them were diagnosed with abdominal ultrasonography. With the growing use of CT in the evaluation of abdominal pain, preoperative diagnosis of the condition may be expedited. CT shows characteristic findings of torsion of the gallbladder such as 1) pronounded swelling of the gallbladder and circumscribed thickening of the gallbladder wall, 2) displacement of the gallbladder, and 3) volvulated gallbladder neck. We found CT to be useful for preoperative diagnosis of patients with abdominal pain