• Title/Summary/Keyword: In situ Hybridization

Search Result 581, Processing Time 0.027 seconds

Application of Molecular Diagnostics Technology in the Development of a Companion Diagnostics for Malignant Solid Tumors (악성 고형암의 항암제 동반진단 기술에서 분자진단기술의 적용)

  • Kim, Jin-Hee
    • The Journal of the Korea Contents Association
    • /
    • v.19 no.3
    • /
    • pp.365-374
    • /
    • 2019
  • Unlike benign tumors, malignant tumors are capable of metastasis, easy to relapse, poor survival, and low quality of life. In Korea, here is a tendency to treat the tumors collectively according to the General Principles of Cancer Chemotherapy(GPCC) of the Health Insurance Review & Assessment Service (HIRA). But recently, companion diagnostics(CDx) is recommended rather than unilateral medication because biomarker-based molecular diagnostics is possible to predict the drug response of patients before drug treatment. Not only domestic but also overseas Food and Drug Administratio (FDA) recommends the development of the CDx system at the stage of drug development to ensure the responsiveness and safety of medicines. In this study, I focused on the necessity of CDx development direction as well as CDx development status through literature review. Furthermore I also discussed CDx types according to the molecular diagnostic technology such as immunohistochemistry (IHC), polymerase chain reaction (PCR), in situ hybridization (ISH), and next-generation sequencing (NGS) not only in the approved CDx but also in the developing one by US FDA. And I suggested the technology issue of CDx development process such as a selection of molecular diagnostics at the time of release, a clear understanding of the CDx mechanism, and a convergence of drug with CDx development. The necessity of social insurance system also was proposed for CDx development.

Comparison of Telomere Length and Vitality among Korean Native Chicken Breeds (토종닭 품종 간 텔로미어 길이 및 생존율 비교 분석)

  • Cho, Eun Jung;Kim, Bo Gyeong;Sohn, Sea Hwan
    • Korean Journal of Poultry Science
    • /
    • v.49 no.1
    • /
    • pp.15-23
    • /
    • 2022
  • We compared the degrees of vitality of 12 Korean native chicken breeds, such as Jaeraejong, Korean Rhode Island Red (Rhode), Korean White Leghorn, Korean Cornish (Cornish), and Korean Ogye breeds. The survival rate and telomere length were measured as vital markers. Telomere length was analyzed via quantitative fluorescence in situ hybridization method using the lymphocytes of 466 chickens. We found that the telomere length decreased linearly with increasing chronological age in all chicken breeds. Telomere length and telomere shortening rates (TSR) were significantly different among the chicken breeds after 20 weeks of age (P<0.01). Rhode had the longest telomere length and the lowest TSR, whereas Cornish had the shortest telomere length and the highest TSR. In terms of TSR, the telomere length of 50-week-old chickens was half of that of 8-week-old chickens. There was also a significant difference in survival rates among the breeds. Both Rhode and Korean Ogye had the highest survival rates, while Cornish had the lowest. There was a significant positive correlation between survival rate and telomere length, and telomere length in old age showed a higher correlation with survival rate than that in young age. Therefore, it is considered that TSR is more closely related to survival rate than the telomere length. Based on the telomere dynamics and survival rates of 12 Korean native chicken breeds, it was concluded that the Rhode breed and Cornish breed had the highest and lowest vitality, respectively.

Biodegradation of VOC Mixtures using a Bioactive Foam Reactor II: Analysis of Microbial Community (계면활성제 미생물반응기의(혼합 VOCs) 생분해 II: 미생물의 군집해석)

  • Jang, Hyun Sup;Shin, Shoung Kyu;Song, Ji Hyeon;Hwang, Sun Jin
    • KSCE Journal of Civil and Environmental Engineering Research
    • /
    • v.26 no.6B
    • /
    • pp.695-701
    • /
    • 2006
  • A toluene-degrading bacterial strain was isolated from a mixed culture that was maintained using toluene as a sole carbon and energy source. The isolated bacterium was classified as Pseudomonas sp. TBD4 based on the close relationship to bacteria belonging to this genus. A bottle study to determine biodegradation rates of individual aromatic compounds showed that the biodegradation was faster in the order of toluene, benzene, styrene, and p-xylene. However, when various mixtures were subjected to TDB4, styrene was degraded at the highest rate, indicating that both toluene and p-xylene could stimulate the degradation of other substrates whereas styrene played as an inhibitor. In addition, the mixed culture and TDB4 were inoculated to the bioactive foam reactor (BFR), and the reactor performance and the corresponding change of microbial community were monitored using the fluorescent in situ hybridization (FISH) method. When an inlet concentration of the VOC mixture increased to greater than 250 ppm, the overall removal efficiency dropped significantly. The FISH measurement demonstrated that the ratio of TDB4 to the total bacteria also decreased to less than 20% along with the decline in removal efficiency in the BFR. As a result, the periodic addition of the pre-grown TDB4 might have been beneficial to achieve a stable performance in the BFR operated over an extended period.

Membranous Nephropathy Associated with Epstein-Barr Virus Infection in a Child (소아에서 엡스타인-바 바이러스 감염과 관련한 막성 신병증 1례)

  • Lee, Eun-Hee;Lim, Dong-Hee;Yim, Hyung-Eun;Yoo, Kee-Hwan;Won, Nam-Hee;Hong, Young-Sook;Lee, Joo-Won
    • Childhood Kidney Diseases
    • /
    • v.12 no.1
    • /
    • pp.88-92
    • /
    • 2008
  • Infection of Epstein-Barr virus(EBV) gives rise to a broad spectrum of clinical manifestations in children. Although renal involvement is rare, diverse renal manifestations are known from hematuria to acute renal failure. Secondary membranous nephropathy(MN) associated with systemic EBV infection is an uncommon renal pathology and only two cases have been reported. We are adding another case of MN associated with EBV infection in a child. An 8-year-old girl was admitted for renal biopsy. She had been followed up for microscopic hematuria and intermittent proteinuria for 5 months. There had been no specific findings in serology and radiology. Tonsil biopsy had been done due to exudative tonsillar hypertrophy and enlarged multiple cervical lymph nodes. And it showed EBV-associated lymphoproliferative findings. Serologic tests for EBV showed positive evidence of recent infection; viral capsid antigen(VCA) IgM was borderline positive, VCA IgG and early antigen IgG were positive, and EB nuclear antigen IgG was negative. In Situ Hybridization of tonsil for EBV mRNA was positive. Because her proteinuria and hematuria were aggravated at that time(protein 3 +, RBC >60/HPF), renal biopsy was done. Renal biopsy showed the findings of MN, characterized by thickened capillary walls with epimembranous spikes on light microscopy and subepithelial, mesangial and subendothelial electron dense deposits on electron microscopy. On immunofluorescence microscopy, IgG, C1q, kappa and lambda chains were positive. After steroid administration, proteinuria and hematuria resolved gradually within 6 months.

  • PDF

The Change of Attached Bacterial Community on Aggregates in Lake Paldang (팔당호에서 Aggregates에 부착한 세균군집구조의 변화)

  • 홍선희;오덕화;전선옥;안태석
    • Korean Journal of Microbiology
    • /
    • v.36 no.4
    • /
    • pp.292-298
    • /
    • 2000
  • For scrutinizing the changes of aggregated bacteria in Lake Paldang, the FISH method was applied by using the rhodamine labeled probes, and total bacteria, chloropyll a concentrations and nutrients concentrations were measured. The aggregates were collected with sediment traps. The T-N, T-P, chlorophyll a concentrations of aggregates were higher 5-15 times, 81-140 and 49-66 times than water samples, respectively. Also, the bacterial numbers of aggregates were 200 times higher than those of water smaples. The ratios of each groups of water sample were 2.1-7.4% for $\alpha$-group, 4.5-8.3% for $\beta$-group, 2.1-7.4% for $\gamma$-group, 2.1-6.1% for Cytophaga-Flavobacterium group and 0.1-2.5% for 'other'group, respectively. While, in aggregates, the ratios of $\alpha$-, $\beta$-, $\gamma$- and Cytophaga-Flavobacterium groups were very small and most abundant group was 'other' bacteria. With these results, the aggregated bacteria in Lake Paldang had a particular group composition of bacteria.

  • PDF

Changes of Nitrifying Bacterial Populations in Anaerobic-Anoxic-Oxic Reactors (혐기-무산소-호기 반응조내 질화세균군의 변화)

  • Park, Jong-Woong;Lee, Young-Ok;Go, Jun-Heok;Ra, Won-Sik;Lim, Uk-Min;Park, Ji-Eun
    • Journal of Korean Society of Environmental Engineers
    • /
    • v.27 no.2
    • /
    • pp.138-144
    • /
    • 2005
  • This study was carried out to investigate the changes of nitrifying bacterial populations including Nitrosomonas sp. and Nitrobacter sp. in $A^2/O$ pilot plant with the configuration of anaerobic-anoxic-oxic reactors. The suspended nitrifying bacterial populations in mixed liquor and those of attached populations on granular carrier surface made by molded waste tire were analyzed by Fluorescent in situ Hybridization(FISH) method. The nitrification rate of a pilot plant showed the value of $1.97{\sim}2.98\;mg\;N/g$ MLVSS hr. The ratios of suspended ammonia oxidizer including Nitrosomonas sp. (NSO) to total bacteria in each reactor were oxic < anoxic < anaerobic. On the contrary, the ratios of suspended nitrite oxidizer including Nitrobacter sp. (NIT) were anaerobic < anoxic < oxic. The thickness, dry density and mass of the attached biomass on granular carriers were $180{\sim}188\;{\mu}m$, $38.5{\sim}43.9\;mg/cm^3$, $29.4{\sim}32.5\;mg/g$, respectively. Also, the ratios of attached nitrifier to total bacteria on granular carriers were similar regardless of ammonia/nitrite-oxidizer (NSO; 3.2%, NIT; 2.8%) and very low compared to those(NSO; $22.8{\sim}28.4%$, NIT; $17{\sim}26%$) of suspended nitrifier.

Detection of HER2 Status in Breast Cancer: Comparison of Current Methods with MLPA and Real-time RT-PCR

  • Pazhoomand, Reza;Keyhan, Elahe;Banan, Mehdi;Najmabad, Hossein;Karimlou, Masoud;Khodadad, Faranak;Iraniparast, Alireza;Feiz, Farnaz;Majidzadeh, Keivan;Bahman, Ideh;Moghadam, Fatemeh Aghakhani;Sobhani, Atoosa Madadkar;Abedin, Seyedeh Sedigheh;Muhammadnejad, Ahad;Behjat, Farkhondeh
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.12
    • /
    • pp.7621-7628
    • /
    • 2013
  • Human epidermal growth factor receptor (HER) status is an important prognostic factor in breast cancer. There is no globally accepted method for determining its status, and which method is most precise is still a matter of debate. We here analyzed HER2 mRNA expression by quantitative reverse transcription-PCR (qRT-PCR) and HER2 DNA amplification using multiplex ligation-dependent probe amplification (MLPA). In parallel, we performed a routine evaluation of HER2 protein by immunohistochemistry (IHC). To assess the accuracy of the RT-PCR and MLPA techniques, a combination of IHC and fluorescence in situ hybridization (FISH) was used, substituting FISH when the results of IHC were ambiguous (2+) and for those IHC results that disagreed with MLPA and qRT-PCR, this approach being termed IHC-FISH. The IHC results for four samples were not compatible with the MLPA and qRT-PCR results; the MLPA and qRT-PCR results for these samples were confirmed by FISH. The correlations between IHC-FISH and qRT-PCR or MLPA were 0.945 and 0.973, respectively. The ASCO/CAP guideline IHC/FISH correlation with MLPA was (0.827) and with RT-PCR was (0.854). The correlations between the IHC results (0, 1+ as negative, and 3+ as positive) and qRT-PCR and MLPA techniques were 0.743 and 0.831, respectively. Given the shortcomings of IHC analysis and greater correlations between MLPA, qRT-PCR, and FISH methods than IHC analysis alone with each of these three methods, we propose that MLPA and real-time PCR are good alternatives to IHC. However a suitable cut-off point for qRTPCR is a prerequisite for determining the exact status of HER2.

Pseudoisodicentric X chromosome in a female with primary amenorrhea (원발성 무월경 여성에서 관찰된 Pseudoisodicentric X 염색체)

  • Park, Sang-Hee;Shim, Sung-Han;Chin, Mi-Uk;Kang, Su-Jin;Bae, Sung-Mi;Sohn, Soo-Min;Cha, Dong-Hyun;Yoon, Tae-Ki;Cho, Jung-Hyun
    • Journal of Genetic Medicine
    • /
    • v.5 no.1
    • /
    • pp.61-64
    • /
    • 2008
  • A 24-year-old female with primary amenorrhea was referred for a chromosome study. The karyotype of the patient was 46,X,der(X) under initial GTG-banding analysis. Fluorescence in situ hybridization (FISH) analysis with an LSI Kallmann (KAL) region probe [probes for Xp22.3(KAL) and CEP(X) for control] was carried out. The abnormal chromosome was KAL- and CEP(X)${\times}2$. In addition, interphase FISH analysis revealed the patient to be mosaic for two different cell lines: 90% of cells had three signals and 10% of the cells had only one signal for CEP(X). Based on these results, the karyotype of the patient was 45,X/46,X,psu idic(X)(p22.1), which is partial trisomy for Xqter${\rightarrow}$Xp22.1 and partial monosomy for Xpter${\rightarrow}$Xp22.1. This karyotype was considered a variant of Turner syndrome. In summary, Idic(X) and low-level mosaicism was successfully characterized by FISH analysis with a CEP(X) probe.

  • PDF

A Case of a 46,XX Male with SRY Gene (SRY 유전자를 가진 46,XX 남성 1례)

  • Min, Jeong-Yong;Lee, Dong-Suk;Cho, Soo-Kyung;Park, So-Hyun;Lee, Soo-Min;Baek, Min-Kyung;Kim, Ki-Chul;Hwang, Do-Yeong
    • Journal of Genetic Medicine
    • /
    • v.5 no.2
    • /
    • pp.145-149
    • /
    • 2008
  • 46,XX male is a rare sex constitution characterized by the development of bilateral testis in persons who lack a Y chromosome. Manifestations of 46,XX males are usually hypogonadism, gynecomastia, azoospermia, and hyalinations of seminiferous tubules. The incidence of XX male reversal is approximately 1 in 20,000 male neonates. The SRYgene is located at the short arm of the Y chromosome(Yp11.31) and codes for testis determining factor in humans. Here, the patient, who presented with a normal male phenotype, was referred for azoospermia. Conventional cytogenetic analysis showed a 46,XX karyotype. Quantitative fluorescent polymerase chain reaction(QF-PCR) and Multiplex PCR studies identified SRY gene. And, Fluorescence In Situ Hybridization(FISH) confirmed the SRY gene on the distal short arm of chromosome X. We identified the SRY gene on the distal short arm of chromosome X by molecular cytogenetic and molecular analyses. Therefore, molecular-cytogenetics and molecular studies were proved to be clinically useful adjunctive tool to conventional prenatal cytogenetic analysis.

  • PDF

Cytogenetic Analyses of Astragalus Species (황기류 식물 3종의 세포유전학적 분석)

  • Kim, Soo-Young;Choi, Hae-Woon;Kim, Chan-Soo;Sung, Jung-Sook;Lee, Joong-Ku;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
    • /
    • v.14 no.4
    • /
    • pp.250-254
    • /
    • 2006
  • To elucidate cytogenetic differences, karyotype analysis and FISH (fluorescence in situ hybridization) with 45S and 5S rDNAs were carried out in the three Astragalas species: Astragalas membranaceus Bunge, A. membranaceus var. alpinus Nakai and A. mongholicus Bunge. The somatic metaphase chromosome numbers of all three species were 2n=2x=16 and the size of chromosomes ranged $2.19{\sim} 5.73\;{\mu}m$. The chromosome complement of A. membranaceus consisted of each four pairs of metacentrics (chromosomes 3,4,6 and 7) and submetacentrics (chromosomes 1,2,4 and 8). In A. membranaceus var. alpinus, the chromosome complement consisted of two pairs of metacentrics (chromosomes 4 and 8) and six pairs of submetacentrics (chromosomes 1,2,3,5,6 and 7). A. mongholicus had three pairs of metacentrics (chromosomes 6,7 and 8) and five pairs of submetacentrics (chromosomes 1,2,3,4 and 5). Using bicolor-FISH, one pair of 45S and 5S rDNA signals could be detected on the centromeric regions of chromosomes 8 and 7 of A. membranaceus and A. mongholicus, respectively. In contrast, A, membranaceus var. alpinus had one pair of 45S signals on the centromeric region of chromosome 8 and two pairs of 5S rDNA signals on the short arms of chromosomes 7 and 8.