• Tuberculosis and Respiratory Diseases
• /
• v.69 no.4
• /
• pp.250-255
• /
• 2010

Background: The purpose of this study was to evaluate recently developed real-time polymerase chain reaction (PCR) assay kit to detect Mycobacterium tuberculosis (MTB) and nontuberculous mycobacteria (NTM) in respiratory specimens. Methods: We assessed the positive rate of the real-time PCR assay to detect MTB and NTM in 87 culture-positive specimens (37 sputum, 50 bronchial washing), which were performed real-time PCR by using $Real-Q_{TM}$ MTB&NTM Kit from January 2009 to June 2009, at Gyeongsang University Hospital. To compare the efficacy with the TB-PCR assay, we evaluated 63 culture-positive specimens (19 sputum, 44 bronchial washing) for MTB or NTM, which were performed TB-PCR by using ABSOLUTETM MTB II PCR Kit from March 2008 to August 2008. Results: Among 87 specimens tested using real-time PCR, MTB and NTM were cultured in 58 and 29, respectively. The positive rate of real-time PCR assay to detect MTB was 71% (22/31) and 92.6% (25/27) in AFB stain-negative and stain-positive specimens. For NTM, the positive rate of real-time PCR was 11.1% (2/18) and 72.7% (8/11) in AFB stain-negative and stain-positive specimens. Among 63 specimens performed using TB-PCR, MTB and NTM were cultured in 46 and 17, respectively. The positive rate of TB-PCR was 61.7% (21/34) and 100% (12/12) in AFB stain-negative and stain-positive specimens. TB-PCR was negative in all NTM-cultured 17 specimens. Conclusion: TB/NTM real-time PCR assay is useful to differentiate MTB and NTM in AFB stain-positive respiratory specimens and it is as effective in detecting MTB with TB-PCR.

• The Journal of the Korean dental association
• /
• v.15 no.8
• /
• pp.611-615
• /
• 1977

The author observed histochemically the nature of the calcified tissue in the rabbit dental pulp, induced by pulpal injection of potassium permanganate. The pulp of rabbit mandibular incisors were exposed and enlarged by a dental hand reamer. The exposed pulps were injected with 0.05ml of 20mM solution of potassium permanganate dissolved in Ringer's solution in experimental tooth. Also the control tooth received a pulpal injection of 0.05ml of Ringer's solution. After pulpal injection, the tooth was plugged with a gutta-percha root canal point. The staining techniques were hematoxylin-eosin stain, van Gieson stain, PAS reaction, toluidine blue stain, alcian blue-hematoxylin stain and colloidal iron-picric acid stain. The results were as follows: 1. The pulp on experimental tooth showed osteodentin-like calcified tissue. Also, in some areas, false denticle-like substance were observed. 2. The central portion of the calcified matrix showed metachromasia in toluidine blue stain had strong staining capacity in alcian blue stain. 3. The peripheral portion of the calcified tissue revealed marked van Gieson positive reaction for collagen. But their staining ability in alcian blue was slight and metachromasia was not appeared.

• The korean journal of orthodontics
• /
• v.25 no.6 s.53
• /
• pp.697-704
• /
• 1995

Chicken calvarial bone is known to contain various cell types, but their exact composition is unknown. By characterizing the chicken calvarial bone biochemically, it can be used to study biochemical, histochemical actions of bone cells in general. Calvaria of 18-day-old white leg horn embryo was aseptically dissected and bone cell populations were isolated by sequential enzymatic digestion. Histochemical study for osteoclast-like bone cell. population was performed with tartrate resistant acid phosphatase(TRAP) stain and for osteoblast-like bone cell population, alkaline phosphatase(ALP) stain was performed. Biochemical study for osteoblast-like bone cell population was performed using alkaline phosphatase(ALP) assay. Following conclusions were obtained from this study. 1. TRAP positive multi and mononuclear cells were mostly observed in group I and II, indicating that osteoclast-like bone cell population is mostly found in these groups. 2. All the cultured groups showed almost equal ALP activities and were positive for ALP stain, indicating that osteoblast-like bone cell population is evenly dispersed in all culture groups. 3. Experimental group treated with $1,25(OH)_{2}D_3$ showed increase in ALP activity in contrast to the control group, confirming previous studies that $1,25(OH)_{2}D_3$ increases ALP activities in in vitro bone cultures. 4. Results from von Kossa's stain indicated that in vitro bone formation had occured after 3 weeks of culture with beta-glycero phosphate.

• Korean Journal of Clinical Laboratory Science
• /
• v.39 no.2
• /
• pp.122-127
• /
• 2007

This study is a report about a specific patient whose primary stomach adenocarcinoma metastasized to uterine cervix adenocarcinoma. A thirty-nine year old female patient was initially diagnosed as having metastatic adenocarcinoma in the supraclavicular lymph node. Upon further examination, she was diagnosed with stomach adenocarcinoma. 8 months later, a cervix punch biopsy was performed. The stains used for examination were H&E stain, PAS stain, Alcian blue stain, Mucicarmine stain, Papanicolaou's (Pap.) stain, and as immunohistochemical stains, cytokeratin 7 and 20 were done. In the H&E stain, the tumor cells showed prominent and eccentric nuclei, thin nuclear membrane in abundant mucous cytoplasm, and cylinder shape. In the PAS stain, intracytoplasmic mucin vacuoles were stained with pink, and in Alcian blue and Mucicarmine stains, intracytoplasmic mucin vacuoles were stained with blue and red. As in the above results, she was diagnosed with undifferentiated adenocarcinoma. As found on the cytologic smear preparation of the uterine cervix stained by Papanicolaou's stains, the background was relatively clear, the number of malignant cells was relatively low, and large and eccentric nuclei in abundant cytoplasm were observed. Upon observing the tissue preparation of the uterine cervix biopsy by H&E stain, a clear background, large and eccentric nuclei, and a signet ring cell types were observed, and the number of malignant cells were fewer than in the primary uterine cervix adenocarcinoma. The vacuoles in cytoplasm were observed. The nuclear membrane and chromatin were thick and very rough, and upon observation by cytokeratin 7 and 20 of immunohistochemical stain, the tumor cells indicated a positive rate of 70% and 20%, respectively. According to these results, also she was diagnosed with metastasized uterine cervix adenocarcinoma. In summary of the results of pathologic findings on stomach biopsy and cytologic, histopathologic, and immunohistochemical finding on uterine cervix biopsy, the adenocarcinoma of her uterine cervix could assert the adenocarcinoma of signet ring cell type that was metastasized from the primary undifferentiated adenocarcinoma in stomach.

• Korean Journal of Poultry Science
• /
• v.26 no.3
• /
• pp.171-177
• /
• 1999

This study was conducted for the histological observation of the infundibulum of the oviduct of the laying Korean native pheasants. The results are as follows : 1. The infundibular wall is composed of the epithelium, lamina propria, muscle layer(inner circular and outer longitudinal muscle), and serosa. The funnel lip is divided into the inner, and outer lip of the epithelium and muscle layer. 2. The epithelium of the funnel lip and most region of the cranial part of the funnel are composed of ciliated columnar cells. In the surface and lateral part of the folds, ciliated cells and non-ciliated secretory cells tend to alternate in the epithelium of the caudal funnel and the necks, but are also found in groups of the simple cuboidal epithelium at the bases of the grooves between the ridges and tubular glands found in the subepithelium. 3. The secretory material of the non-ciliated secretory cills contains PAS-positive and alcian blue-positive granules, and these materials show purple colour in the basic fuchsin-methylene blue stain. 4. The cells of the glandular groove and tubular gland of the neck portion of the oviduct mostly show weak PAS-positive, and alcian blue stain negative reaction. The tubular gland cells of the infndibulum contain pink of purple colour granules, and without reaction in the anterior neck portion of the infundibulum in basic fuchsin and methylene blue stain.

• Applied Microscopy
• /
• v.24 no.4
• /
• pp.107-114
• /
• 1994

The Rotaviruses are members of the family Reoviridae and are the major cause of severe childhood gastroenteritis worldwide. Recently, electron microscopy has been used to detect non-group A rotaviruses to determine a relatively high resolution structure of the rotavirion. Mature, infectious virions(double-shelled particles) have a diameter of approximately 70nm, and have a capsid structure composed of two concentric protein layers. We have studied patient's stool specimen by negative staining technique complete removal of sucrose suspension. This negative staining technique that could be carried out in about 30 minutes and that could be used with crude stool specimen was an advantage of major significance. Removal of sucrose in the sample by has been completed washing with distilled of sucrose and by washing with distilled water. Ultrastructurally, typical feature of rotavirus has a double capsid construction with an inner capsid of 55nm and on outer 65-70nm diameter can be clearly demonstrated.

• Biomedical Science Letters
• /
• v.19 no.2
• /
• pp.153-157
• /
• 2013

Although culture is the gold standard method to identify mycobacteria, its use in tuberculous lymphadenitis (TBL) is limited due to formalin fixation of the submitted specimens. We evaluated the performance of quantitative real-time PCR (q-PCR) for Mycobacterium Tuberculosis (MTB) in granulomatous lymphadenitis using formalin-fixed paraffin-embedded (FFPE) tissues. From 2000 to 2010, a total number of 117 cases of lymph node samples with granulomatous inflammation which were surgically removed and fixed in formalin were studied. Hematoxylin & Eosin (H&E) and Ziehl-Neelsen-stained (ZN) slides were reviewed. qPCR using Real TB-Taq$^{(R)}$ was performed for all cases to identify Mycobacterium tuberculosis. Thirteen non-tuberculous lymphadenopathy cases were used as negative control. Cervical lymph nodes were more frequently affected (60%, 70/117) than other sites. ZN stain for acid fast bacilli was positive in 19 (16.24%) cases. qPCR for tuberculosis was positive in 92 (78.63%) cases. Caseous necrosis was found in 103 (88.03%) cases. While the ZN stain and qPCR were both negative in all control cases, the qPCR showed a significantly higher positive rate (78.63% vs. 16.24%) compared to ZN stain in histologically diagnosed TBL. Quantitative real-time PCR proves to be more sensitive than ZN stain for diagnosis of tuberculous lymphadenitis.

• Journal of Species Research
• /
• v.7 no.3
• /
• pp.222-230
• /
• 2018

Six bacterial strains 18JY42-3, 18SH, 18JY76-11, 17J11-11, 18JY14-14, and 18JY15-11 assigned to the phylum Proteobacteria, Firmicutes, and Actinobacteria were isolated from soil samples in Korea. The Cohnella species, strain 18JY42-3 was Gram-stain-positive, short rod-shaped and beige-colored. The Methylobacterium species, strains 18SH and 18JY76-11 were Gram-stain-negative, short rod-shaped and pink-colored. The Microterricola species, strain 17J11-11 was Gram-stain-positive, short rod-shaped and yellow-colored. The Paenarthrobacter species, strains 18JY14-14 and 18JY15-11 were Gram-stain-positive, short rod-shaped and white-colored. Phylogenetic analysis based on 16S rRNA gene sequence showed that strains 18JY42-3, 18SH, 18JY76-11, 17J11-11, 18JY14-14, and 18JY15-11 were most closely related Cohnella rhizosphaerae (MH497628; 98.8%), Methylobacterium goesingense (MH497632; 99.1%), Methylobacterium populi (MH497635; 99.9%), Microterricolagilva (MH504108; 98.4%), Paenarthrobacter nicotinovorans (MH497641; 100%), and Paenarthrobacter nitroguajacolicus (MH497646; 99.2%), respectively. All the six unrecorded strains showed resistance to UV radiation. This is the first report of these six species in Korea.

• Korean Journal of Microbiology
• /
• v.25 no.3
• /
• pp.244-248
• /
• 1987

Attempts were made to enumerate the number of Gram positive and negative bacteria in the development of natural fermentation rapidly and simultaneously. A general Gram stain was applied to this study. The number of cells by Gram stain was proportional to the cell turbidity by spectrophotometer within a range of 0.7 absorbance at 610nm. The cells washed out during procedures were not exceeded about 8 percentage. The standard error of separate counts in the mixture of Cscherichia coli and Micrococcus luteus was $5.1\pm2.3$%. The possible range of counting was $5.5\times 10^{7}-1.0\times 10^{9}$ cells/ml. Therefore, it is believed that a general Gram stain could be applied to the separate counting of mixture of Fram positive and negative bacterial populations too. In practice, growth kinetics of hemp retting and Kimchi fermentation were presented.

• Parasites, Hosts and Diseases
• /
• v.50 no.1
• /
• pp.57-62
• /
• 2012

A total of 450 stool samples were collected from inpatient and outpatient clinics of Pediatric Department, Minia University Hospital, Minia District, Egypt. Two groups of patients were studied, including 200 immunosuppressed and 250 immunocompetent children. Stool samples were subjected to wet saline and iodine mounts. A concentration technique (formol-ether sedimentation method) was carried out for stool samples diagnosed negative by wet saline and iodine mounts. Samples were stained by 2 different methods; acid fast stain (modified Ziehl-Neelsen stain) and Giemsa stain. Total 188 cases (94%) were diagnosed positive for parasitic infections among immunosuppressed children, whereas 150 cases (60%) were positive in immunocompetent children ($P$<0.0001). The most common protozoan infection in immunosuppressed group was $Cryptosporidium$ $parvum$ (60.2%), followed by $Blastocystis$ $hominis$ (12.1%), $Isospora$ $belli$ (9.7%), and $Cyclospora$ $caytenensis$ (7.8%). On the other hand, $Entamoeba$ $histolytica$ (24.6%) and $Giardia$ $lamblia$ (17.6%) were more common than other protozoans in immunocompetent children.