• Tuberculosis and Respiratory Diseases
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• v.48 no.5
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• pp.699-708
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• 2000

Background : A presumptive histopathologic diagnosis of tuberculosis is commonly based on the finding of acid-fast bacilli upon microscopic examination of a diagnostic specimens. Although this traditional histochemical staining method is satisfactory, it is time-consuming and not species-specific. For more specific assessment, in situ hybridization assay with oligonucleotide probes is introduced. Methods : The human surgical specimens were obtained from tuberculosis patients, and experimental specimens were made by injecting cultured M. tuberculosis organisms into fresh rat liver. Oligonucleotide probes complementary to ribosomal RNA portion were synthesized and labeled with multiple biotin molecules. For a rapid detection, all procedures were carried out using manual capillary action technology on the Microprobe staining system. Results : The in situ hybridization assay produced a positive reaction in experimental specimens (80-90% sensitivity) after pepsin-HCl pre-treatment for a good permeabilization of probes, but reliable result was not obtained from human surgical specimens. Conclusion : It is, therefore, suggested that biotin-labeled oligonucleotide probes have considerable potential for identification and in situ detection of M. tuberculosis but, there are some barriers to overcome for the diagnostic use of this method.

• Journal of the korean academy of Pediatric Dentistry
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• v.34 no.2
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• pp.299-308
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• 2007

Insoluble glucan is the important component of oral biofilm, which is synthesized from sucrose through the action of glucosyltransferase (GTF) B and GTF C encoded by the gtfB and gtfC genes, respectively of Streptococcus mutans. In present study, the effects of various sugars on the mRNA expression of gtfB and gtfC of S. mutans Ingbritt were examined by fluorescent in situ hybridization (FISH). The mRNA of gtfB and gtfC was expressed normally in the BHI broth containing 1% and 5% sucrose. The mRNA expression was decreased by the addition of 10% of glucose, and 1%, 5% and 10% of fructose. Lactose had no great effect on the expression of gtfB and gtfC. 5% and 10% of xylitol greatly reduced the mRNA expression of gtfB and gtfC. Sorbitol slightly decreased the mRNA expression of gtfB and gtfC when compared to the control. In summary, mRNA expression of gtfB and gtfC was decreased by the addition of glucose, fructose, and xylitol.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.2
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• pp.829-834
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• 2013

Background: MicroRNAs are noncoding RNA molecules that posttranscriptionally regulate gene expression. The aim of this study was to determine the role of microRNA-21 in cholangiocarcinomas and its relationship to cholangiocarcinoma RBE cell capacity for invasion and metastasis. Methods: MicroRNA-21 expression was investigated in 41 cases of cholangiocarcinoma samples by in situ hybridization and real-time PCR. Influence on cholangiocarcinoma cell line invasion and metastasis was analyzed with microRNA-21 transfected cells. In addition, regulation of reversion-inducing-cysteine-rich protein with kazal motifs (RECK) by microRNA-21 was elucidated to identify mechanisms. Results: In situ hybridization and real-time quantitative PCR results for patients with lymph node metastasis or perineural invasion showed significantly high expression of microRNA-21 (P<0.05). There was a dramatic decrease in cholangiocarcinoma cell line invasion and metastasis ability after microRNA-21 knockdown (P<0.05). However, overexpression significantly increased invasion and metastasis (P<0.05). Real-time PCR and Western-blot analysis showed that microRNA-21 could potentially inhibit RECK expression in RBE cells. Survival analysis showed that patients with higher expression levels of microRNA-21 more often had a poor prognosis (P<0.05). Conclusions: MicroRNA-21 may play an important role in cholangiocarcinoma invasion and metastasis, suggesting that MicroRNA-21 should be further evaluated as a biomarker for predicting cholangiocarcinoma prognosis.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.282-285
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• 2001

The microbial community structure and in situ spatial distribution of ammonia oxidizing and nitrite oxidizing bacteria in nitrifying biofilm of an upflow biological aerated filter system were investigated. The reactor had been continuously operated under high free ammonia concentration and low DO concentration for nitrite accumulation more than 2 years before the experiment. Fluorescence in situ hybridization

• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.299-307
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• 2003

Objectives: The present study was conducted to evaluate the mobile transposon-like element, clone MTi7 (MTi7) expression in the mouse ovary and to determine its role(s) in the mouse oocytes by RNA interference (RNAi). Methods: MTi7 mRNA expression was localized by in situ hybridization in day5 and adult ovaries. Double stranded RNA (dsRNA) was prepared for c-mos, a gene with known function as control, and the MTi7. Each dsRNA was microinjected into the germinal vesicle (GV) stage oocytes then oocyte maturation and intracellular changes were evaluated. Results: In situ hybridization analysis revealed that MTi7 mRNA localized to the oocyte cytoplasm from primordial to preovulatory follicles. After dsRNA injection, we found 43-54% GV arrest of microinjected GV oocytes with 68%-90% decrease in targeted c-mos or MTi7 mRNA. Conclusions: This is the first report of the oocyte-specific expression of the MTi7 mRNA. From results of RNAi for MTi7, we concluded that the MTi7 is involved in the germinal vesicle breakdown in GV oocytes, and MTi7 may be implicated with c-mos for its function. We report here that RNAi provides an outstanding approach to study the function of a gene with unknown functions.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.5
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• pp.361-367
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• 2000

Cocaine functions as indirect dopamine and serotonin (5-hydroxytryptamine, 5HT) agonists and induces genomic and behavioral alterations in the striatum. Previously we demonstrated that ritanserin, a 5HT2/1C receptor antagonist, is not responsible for cocaine-induced behavioral alterations and zif268 mRNA gene expression in the striatum (see the previous paper in this issue). In this study, it was hypothesized that dopamine and 5HT2/1C receptors are required for cocaine-induced behavioral alterations and c-fos and zif268 mRNA expression. This hypothesis was addressed by infusing amperozide which antagonizes both 5HT2/1C and dopamine receptors and was analyzed using the quantitative in situ hybridization histochemistry in vivo. Systemic injection of amperozide (5 mg/kg, s.c.) significantly blocked increase in behavior, c-fos and zif268 mRNA expression induced by 15 mg/kg cocaine, i.p., in the dorsal striatum. These data suggest that dopamine and 5HT2/1C receptors are necessary for cocaine-induced behavioral alterations and immediate early gene expression in the dorsal striatum.

• Korean Journal of Veterinary Research
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• v.40 no.2
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• pp.229-236
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• 2000

The synthesis of steroid hormone starts from cholesterol. Steroidogenic acute regulatory protein (StAR) acutely transfers cholesterol from the outer mitochondrial membrane to the inner in the early step of steroidogenesis. Many kinds of steroid hormone are mainly synthesized in adrenal grand, ovary, and testis. Among the steroid hormone, testosterone is synthesized in Leydig cells of the testis, the production of testosterone significantly increases in adult testis after puberty onset. Therefore, we think that the expression of StAR mRNA in testis will change according to the testicular development. The aim of this study is to determine the distribution of StAR mRNA in immature and adult rat testes and to confirm the functions of StAR in these testes. Thus, in situ hybridization was used in rat testes of the 2, 4, and 10 weeks of age. StAR mRNA was expressed in Leydig cells. Positive signals of StAR mRNA were weakly detected in Leydig cells of the 2 weeks of age. But, StAR mRNA was strongly expressed in Leydig cells of the 4 and 10 weeks of age, where steroidogenesis actively occur. In our results, the pattern of StAR mRNA expression was similar to the pattern of testosterone production in immature and adult rat testes. In conclusion, we can suggest that StAR acts as an important factor to regulate the synthesis of testosterone in Leydig cells of the rat testis.

• Journal of the korean academy of Pediatric Dentistry
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• v.31 no.2
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• pp.304-313
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• 2004

Xylitol, a five-carbon natural sugar alcohol, is widely used non-cariogenic sugar substitute. In present study, the effects of xylitol on the expression of mRNA for glucosyltransferase which synthesizes glucan from sucrose were detected by Fluorescent in situ hybridization (FISH) and flow cytometry. FITC fluorescences for mRNA of gtfB, gtfC and gtfD were decreased further with increasing concentration of xylitol from 1% to 10% when detected by FISH. Flow cytometric analysis also showed that the expression of gtfB, gtfC and gtfD was increased by the addition of sucrose and decreased by the addition of xylitol to BHI broth containing 1% sucrose. In conclusion, the expression of gtfB, gtfC and gtfD mRNA was decreased by the addition of xylitol.

• Parasites, Hosts and Diseases
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• v.34 no.3
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• pp.177-184
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• 1996

In situ hybridization was performed to detect rat heumocwstis ca4nii in the lung sections. Rats were immunosuppressed by weekly subcutaneous injection of 10 mg/kg methylprednisolone. On the 6th, 8th and 9th week of immunosuppression, the lungs were removed and fled in 10% neutral formalin. A 22 base oligonucleotide probe complementary to p. carinii 5S ribosomal RMh was commercially synhesized and its 3' terminal was labeled wiH biotin. In situ hybridization was performed utilizing manual capillary action technolog)r on the Microprobe system. p. cnrinii were detected along the luminal surface of alveolar pneumocytes, in exudate of alveolar cavities, and also in secretory material of bronchioles. In the 6th week group, positive reaction was observed focally in the peripheral region of the lung sections, but the reaction was observed diffusely in the 8th or 9th week groups. In comparison with Grocott's methenamine silver stain, in situ hybridization technique can detect the organism rapidly, and can detect trophic forms very well. Furthermore, no nonspecific reaction with other pathogenic fungi and protozoa was recognized. Therefore, in situ hybridization can be a good technique to detect p. carinii in the lungs of infected rats.

• Development and Reproduction
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• v.2 no.1
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• pp.39-43
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• 1998

The synthesis of steroid hormone starts from cholesterol. Steroidogenic acute regulatory protein(StAR) transfers cholesterol acutely from the outer mitochondrial membranes to the inner in the early step of steroidogenesis. Many kinds of steroid hormones are mainly synthesized in adrenal grand, ovary and testis. The purpose of this study is to determine the distribution of StAR mRNA in the rat ovary and adrenal gland and to confirm the functions of StAR in these organs. In the ovary, StAR mRNAs were strongly expressed in the corpus luteum, where progesterone is synthesized, and these were weakly expressed in the theca layer of follicles, where androgen is synthesized. However, StAR mRNAs were not detected in the estrogen producing granulosa cells of growing follicles. In the corpus luteum, StAR mRNAs were strongly loclized in the zona fasciculata and zona reticularis, where glucocorticoid is mainly synthesized. StAR mRNAs were weakly expressed in the zona gromerulosa, where mineralcorticoid is synthesized. StAR mRNAs were not detected in the adrenal medulla. In our results, StAR mRNAs were expressed differentially in the steroidogenic cells of ovary and adrenal gland according to the types of steroid hormones, and the statges of corpus luteum development. We conclude that StAR is involved in the steroidogenesis at the very early step of steroid synthesis cascade.