• Title/Summary/Keyword: Negative Stain

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Concurrent occurrence of apocrine sweat gland adenocarcinoma and mammary gland complex adenoma in a dog (개에서 땀샘 선암종과 유선 복합형 선종의 동시 발생)

  • Park, Ji-Sung;Jung, Ji-Youl;Jo, Suk Hee;Cheong, Jongtae;Kang, Tae-Young;Kim, Jae-Hoon
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.311-316
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    • 2008
  • A 7-year-old female Shih Tzu dog with lots of masses in the whole mammary gland was presented to the surgery department of the Veterinary Teaching Hospital in the Cheju National University. After surgical excision, all mammary samples were referred to Pathology Department of Veterinary Medicine. Grossly, masses were measuring up to $6.5{\times}4{\times}1cm$ and on cut surface of masses in right 1st, 3rd, 4th, 5th and left 1st, 3rd, 4th, 5th mammary masses were well delineated and firm, sulphur yellow, solid round to oval shape. Microscopically, most neoplastic sweat glands were severely proliferated in dermis and subcutis. Most tubules were lined by round to oval shaped epithelium with eosinophilic cytoplasm, hyperchromatic nuclei with high mitotic figures and severe central necrosis. The neoplastic epithelium also had PAS-positive diastase-resistant cytoplasmic granules, but negative with Perls iron stain. The left 2nd mass was well delineated, and had several dark brown areas and yellowish white glittered areas. Mass was well circumscribed with dense connective tissue. Neoplastic areas contained irregular sized mammary gland with papillary grown luminal epithelial cells in single or double cells layer with mitotic figures and small amounts of proliferated myoepithelial cells. Proliferated myoepithelial cells also produced slightly basophilic mucinous materials. Based on the gross, histopathologic and special staining characteristics, this dog was diagnosed as 90% of apocrine sweat gland aenocarcinoma and 10% mammary. complex adenomas in mammary masses. In our best knowledge, this is the first report for concurrent occurrence of apocrine sweat gland adenocarcinoma and mammary gland complex adenoma in mammary masses of the same dog.

Hemozoin Pigment: An Important Tool for Low Parasitemic Malarial Diagnosis

  • Mohapatra, Sarita;Ghosh, Arnab;Singh, Ruchi;Singh, Dhirendra Pratap;Sharma, Bhawna;Samantaray, Jyotish Chandra;Deb, Manorama;Gaind, Rajni
    • Parasites, Hosts and Diseases
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    • v.54 no.4
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    • pp.393-397
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    • 2016
  • Low parasitemic condition in malaria remains a diagnostic challenge; as the available diagnostic methods failed to detect. Currently, hemozoin (Hz) pigment is gaining attention in the diagnosis of malaria. The major drawback is ease of detection of Hz in routine practice. A pilot study was conducted to evaluate the role of Hz pigment and to compare the performance of quantitative buffy coat assay (QBC) and PCR in such conditions. Clinically suspected cases of malaria were examined by both Giemsa stain and immunochromatographic test (ICT). Samples positive by ICT and negative by Giemsa stain were further examined by nested PCR targeting 18S rRNA and QBC for the presence of malaria parasites and pigments. Thirty blood samples fulfilled the inclusion criteria out of which 23 were Plasmodium vivax (Pv), 4 Plasmodium falciparum (Pf), and 3 mixed (Pv and Pf) by immunochromatographic test. Twenty-one out of 30 (70%) were positive by nested PCR in comparison to 25/30 (83%) by QBC. Samples containing both malaria parasites and Hz pigment by QBC completely showed concordance with the PCR result. However, 61% of total samples containing only Hz pigment were observed positive by PCR. Hz pigment remains an important tool for malaria diagnosis. Identification of leukocytes containing pigments by QBC not only indicates recent malarial infections but also puts light on severity of the disease. QBC assay is a rapid, highly sensitive, and cost-effective method to detect malaria parasites and Hz pigment especially in low parasitemic conditions.

Studies on antigenicity and production of monoclonal antibody for diagnosis of canine heartworm(Dirofilaria immitis) (개 심장사상충(Dirofilaria immitis) 진단을 위한 항원성 조사 및 단크론항체 생산)

  • Lee, Cheol-soon;Jee, Cha-ho
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.130-137
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    • 2000
  • In order to diagnose canine heartworm infection by antigen capture ELISA, the crude somatic(S), partial somatic(below 45kDa) and excretory/secretory(E/S) antigen of adult heartworm were identified and the antigenicity was examined by silver stain, immunoblot and ELISA. Then, production of monoclonal antibody to specific antigen carried out in this experiment. The bands to S antigen and E/S antigen were recognized between 10 and 200kDa and common bands were recognized strongly 14, 18, 28, 43kDa by silver stain. By western blot analysis, fractions to S antigen were recognized 14, 16, 18, 20, 24, 28, 32, 43, 50, 55kDa, etc. and only a 14kDa to E/S antigen in positive sera which were positive in modified Knott's test and necropsy. In ELISA, the positive sera reacted to antigens(SA, $SA_{45}$, E/S) were significantly different from negative sera by Student's t-test(p<0.05). Four hybridoma cell lines(14, 16, 17, 32kDa) than produce specific monoclonal antibodies for these antigens were obtained by immunizing BALB/c mice with a partially purified somatic antigen (below 45kDa) preparation, by fusing spleen cells with SP2/O cell myeloma cells, and by screening cell culture supernatants for antibody. In these results, it was confirmed that partial somatic antigen(below 45kDa) or E/S antigen can be used for serologic diagnosis of heartworm infection and monoclonal antibody reacting with specific antigen(14kDa) can be used for antigen capture ELISA in prepatent period of canine heartworm infection.

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Apocrine sweat gland adenocarcinoma in a cat (고양이의 땀샘 선암종 증례)

  • Park, Min-Hyeok;Jung, Ji-Youl;Jo, Suk-Hee;Kim, Jae-Hoon;Lee, Jin-Yong;Lee, Mi-Kyung;Kim, Jae-Hoon
    • Korean Journal of Veterinary Research
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    • v.49 no.2
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    • pp.157-161
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    • 2009
  • A 11-year-old female mixed cat with subcutaneous mass around the left 5th mammary glands was presented to local animal hospital. According to history taking, the mass recurred 2 times on the same site of abdomen. After surgical excision, subcutaneous mass was referred to Pathology Department of Veterinary Medicine in the Jeju National University. Grossly, round to oval, milky yellow or pale red nodules, measuring 0.1${\sim}$1 cm in diameter, were occupied in the subcutis. Microscopically, the most neoplastic sweat glands were proliferated in the dermis and subcutis. Most tubules were lined by round to oval shaped epithelium with eosinophilic cytoplasm, hyperchromatic nuclei with high mitotic figures and severe central necrosis. The neoplastic epithelium also had periodic acid-Schiff-positive diastase-resistant cytoplasmic granules, but was negative for Perl's iron stain. Based on the gross, histopathologic and special staining, this cat was diagnosed as apocrine sweat gland adenocarcinoma. In our best knowledge, this is the first report of apocrine sweat gland adenocarcinoma around abdominal mammary gland in a cat.

Non-hemolytic, Mucinous, Coagulase Negative MRSA Isolated from Urine (소변에서 분리된 비용혈성, 점액성, 응고효소 음성 MRSA)

  • Kim, Jae Soo;Choi, Qute;Jung, Bo Kyeung;Kim, Jong Wan;Kim, Ga Yeon
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.2
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    • pp.260-264
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    • 2019
  • An 84-year-old woman presented to the emergency department with a chief complaint of pressure sores of the anus. She had a urine catheter when she showed pyuria three times but had no fever. A microscopic examination revealed many grapevine-like Gram positive strains and neutrophils. After 24 hours of urine culture on blood agar, non-hemolytic mucous colonies were found and further enlarged after 48 hours of culture. The capsules were identified after India ink stain. The catalase was positive, but the tube coagulase and latex coagulase were both negative. The S. aureus was identified by Vitek-2 and mass spectrometer Vitek MS V-3 IVD. The strain was confirmed by 16S rRNA gene sequencing and multilocus sequence typing (MLST). The phenotypically atypical MRSA found in the tube coagulase and latex coagulase were both negative. MRSA often show no beta hemolysis as in this case but are rarely latex coagulase-negative. We report a woman whose urine culture showed non-hemolytic, tube coagulase-negative, and latex coagulase-negative MRSA.

Development of Semen Transport System for Cryopreservation and Fertility in Bull Sperm (소 정자의 동결 및 수정능력 향상을 위한 정액운반법의 개발)

  • Lee, Sang-Hee;Song, Eun-Ji;Woo, Jea-Seok;Lee, Seung-Hwan;Kang, Hee-Seol;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.97-102
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    • 2013
  • The objective of this study was to develop of semen transport system for cryopreservation and fertility in bull sperm. The ejaculated semen were diluted with Triladyl containing 20% egg-yolk for transportation. Diluted semen was transported by three methods that there were wrapping tissue (Tissue), sinking under $30^{\circ}C$ water (Water) and sinking between warm water and air (Air) methods. Semen was transported within 2 hours in $0.3^{\circ}C$. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk. And frozen-thawed sperm were estimated with SYBR14/PI double stain for viability, FITC-PNA/PI double stain for acrosome reaction analysis and Rhodamine123 double stain for mitochondrial intact assessment. In results, live sperm (SYBR+/PI-) in Air treatment group ($43.3{\pm}4.7%$) was significantly (p<0.05) higher than other treatment groups (Tissue: $16.3{\pm}2.7%$ and Water: $27.5{\pm}3.1%$), dying sperm (SYBR+/PI+) in Air treatment group ($55.6{\pm}4.7%$) was significantly lower than other treatment groups (Tissue: $77.6{\pm}3.2%$ and Water: $67.6{\pm}3.3%$) (p<0.05). Acrosome reaction in Air treatment group ($0.2{\pm}0.1%$) within live sperm (PI negative region) was significantly (p<0.05) lower than other treatment groups (Tissue: $0.7{\pm}0.2%$ and Water: $0.5{\pm}0.1%$), the acrosome reaction in Air treatment group ($28.6{\pm}2.8%$) within all sperm also was significantly lower than other treatment groups (Tissue: $44.2{\pm}1.8%$ and Water: $36.2{\pm}2.0%$) (p<0.05). And mitochondrial intact in Air treatment group within live ($97.1{\pm}0.4%$) and all ($61.9{\pm}3.3%$) sperm were significantly higher than other treatment groups (Tissue: $85.2{\pm}3.3%$, Water: $87.8{\pm}2.9%$ within live sperm and Tissue: $49.28{\pm}3.7%$, Water: $42.0{\pm}3.1%$ within all sperm) (p<0.05). Therefore, we suggest that transportation by sinking method between warm water and air was beneficial to improvement of fertility in frozen-thawed in bull semen.

Identification of Daphne Mottle Virus Isolated from Daphne odora, a New Member of the Genus Potyvirus (서향에서 분리한 신종 포티바이러스(Daphne Mottle Virus)의 동정)

  • Park, Chung Youl;Park, Jungan;Lee, Boo-Ja;Bak, Sangmin;Lee, Hong-Kyu;Kim, Jeong-Sun;Yoon, Youngnam;Suh, Sang Jae;Lee, Su-Heon
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.59-63
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    • 2016
  • A new poty-like virus was isolated from plants of winter daphne (Daphne odora) that showed virus-like symptoms on leaves, from four regions of Korea during 2014. Filamentous-shaped particles were observed by transmission electron microscopy of preparations extracted from symptomatic leaves and examined by the direct negative stain method. RT-PCR assay showed that three samples were positive for both Cucumber mosaic virus and potyvirus, and only one sample was positive for potyvirus only. A BLAST comparison to partial sequences from helper-component proteinase, cylindrical inclusion and coat protein genes detected the highest nucleotide identity of 76%, 72%, and 72% with Daphne mosaic virus, respectively, levels below the potyvirus species discrimination threshold. The new potyvirus was isolated using indicator plants (Chenopodium amaranticolor), in which local lesions were produced. In this study, we identified a novel potyvirus from winter daphne, which we have named Daphne mottle virus (DapMoV).

A Clinical Trial of Dextranase-Containing Mouthwash on the Inhibition of Plaque Formation and Gingivitis (Dextranase 함유 구강 세정액의 치태 억제 및 치은염 예방 효과에 관한 임상적 연구)

  • Song, Woo-Sung;Son, Eun-Ju;Kim, Do-Man;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.31 no.2
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    • pp.371-388
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    • 2001
  • A novel glucanhydrolase(DXAMase) from a mutant of Lipomyces starkeyi(KSM 22) has been shown effective in hydrolysis of mutan, reduction of mutan formation by Streptococcus mutans and removal pre-formed sucrose-dependentadherent microbial film and DXAMase has been strongly bound to hydroxyapatitie. These in vitro properties of Lipomyces starkeyi DXAMase are desirable for its application as a dental plaque control agent. This study was performed to determine the adjunctive oral hygiene benefits and safety of dextranase(Lipomyces starkeyi KSM 22 DXAMase)-containing mouthwash when used alongside normal tooth-brushing. This 6-month clinical trial was placebo-controlled double-blind design evaluating 1U/ml dextranase mouthwash and 0.12% chlorhexidine mouthwash. A total 39 systemically healthy subjects, who had moderate levels of plaque and gingivitis were included. At baseline, 1, 3 and 6 months, subjects were scored for plaque accumulation(Turesky modification of Quingley-Hein's plaque index), gingivitis status($L\ddot{o}e$ and Silness gingival index), and tooth stain(Area and severity index system by Lang et al). Additionally, oral mucosal examinations were performed and subjects questioned for adverse symptoms. Two weeks after pre-experiment examinations and a professional prophylaxis, the subjects provided with allocated mousewash and instructed to use 20-ml volumes for 30s twice daily after toothbrushing. All the groups showed significant increase in plaque accumulation since 1 month of experiment. During 6 months' period, the Dextranase mouthwash group showed the least increase in plaque accumulation, compared to the Chlorhexidine mouthwash and placebo groups. As for gingival inflammation, all the groups showed significant increase during 6 months of experiment. The Experimental group(Dextranase mouthwash) also showed the least increase in gingival index score, compared to the Positive control(Chlorhexidine mouthwash)as well as the Negative control(placebo)groups. Whereas the tooth stain was increased significantly in the Positive control group, compared to the baseline score and the Negative controlgroup since 3 months of mouthrinsing. It was significantly increased after 6 months in the Experimental group, still less severe than the Positive control group. As for the oral side effect, the Experimental group showed less tongue accumulation, bad taste, compared to the Positive control group. From these results, mouthrinsing with Lipomyces starkeyi KSM 22 dextranase provided adjunctive benefits to toothbrushing, comparable to 0.12% chlorhexidine mouthwash in inhibition of plaque accumulation and gingival inflammation and local side effects were if anything less frequent and less intense than chlorhexidine, with long-term use of the mouthwash. All data had provided positive evidence for Lipomyces starkeyi KSM 22 dextranase as an antiplaque agent and suggested that further development of dextranase formulations for plaque control are warranted.

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Isolation and comparison of somatic and excretory-secretory antigens for serological diagnosis in Trichuris suis (혈청학적 진단을 위한 돼지 편충의 체항원, 배설/분비항원의 분리 및 비교)

  • Jee, Cha-ho;Lee, Chul-soon;Park, Seung-jun
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.159-168
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    • 1999
  • Swine whipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swine gastrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the reasonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follows; 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values($mean{\pm}SD$) of adult somatic antigen against positive(negative) sera were $0.30{\pm}0.12(0.09{\pm}0.006)$ and third-stage larva of somatic antigen were $0.28{\pm}0.038(0.10{\pm}0.005)$. And OD values of excretory-secretory antigens of adult and third-stage larva were $0.24{\pm}0.031(0.11{\pm}0.005)$ and $0.08{\pm}0.013(0.10{\pm}0.003)$, respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p<0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were $0.30{\pm}0.120(0.09{\pm}0.006)$ and $0.25{\pm}0.141(0.09{\pm}0.003)$. These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.

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Hormonal Activity of Ovarian Fibroma and Thecoma (난소 섬유종과 포막종의 호르몬 분비능에 관한 연구)

  • Lee, Soong-Deok;Suh, Jin-Suk;Han, Young-Mee;Kim, Jung-Ran;Seo, Jeong-Wook;Ham, Eui-Keun;Lee, Hyo-Pyo
    • Applied Microscopy
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    • v.19 no.1
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    • pp.27-33
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    • 1989
  • Six cases of ovarian fibrous stromal neoplasm were studied clinically, light microscopically and ultrastructurally for the clinico-pathological evidences of hormone production. Of the six cases, two cases were fibroma, three cases were fibrothecoma, and one case was thecoma. Two cases of fibroma and one fibrothecoma were associated with clinical history of menstrual abnormality, however fat staining of the tumor was negative or weakly positive. Two cases of fibrothecoma and one thecoma were negative for the clinical history of hormone imbalance. Fat stain of those cases revealed positive in varying intensity. Ultrastructural examination of fibroma-thecoma group revealed dark and pale cells by their nuclear characteristics. The dark cells had indented nucleus and abundant cytoplasmic organelles of rough ER, Golgi apparatus and mitochondria. Intracytoplasmic cisternal spaces were seen in the dark cell cytoplasm and some lipid droplets were seen around the cisternae. Pale cells had pale swollen nucleus and fine chromatins. Their cytoplasm showed scanty amount of organelles. Fibroma-thecoma spectrum showed varying degree of population of dark cells, light cells and intervening collagenous stroma. Lipid droplet was structurally associated with intracytoplasmic cisterna and they were frequently seen in thecoma and two of the fibrothecoma. But clinical history of hormone imbalance was poorly related to the light microscopic morphology and ultrastructural organization.

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