• Title/Summary/Keyword: paraffin-embedded specimens

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Prognostic Significance of Human Epidermal Receptor (HER)-3 Immunohistochemical Expression in Patients with Metastatic Breast Cancer

  • Olmez, Omer Fatih;Evrensel, Turkkan;Cubukcu, Erdem;Ugras, Nesrin;Avci, Nilufer;Canhoroz, Mustafa;Deligonul, Adem;Hartavi, Mustafa;Olmez, Fatma;Cubukcu, Sinem;Tolunay, Sahsine;Kurt, Ender;Kanat, Ozkan;Manavoglu, Osman
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4115-4119
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    • 2013
  • Background: Previous reports have shown that human epidermal receptor (HER)-3 overexpression may be associated with poor prognosis in patients with breast cancer, but results have been conflicting. In this study, we sought to investigate the prognostic significance of HER-3 immunohistochemical expression in patients with metastatic breast cancer. Methods: We retrospectively analyzed HER-3 immunohistochemical expression profiles in 45 paraffin-embedded specimens from patients who had been treated between 1996 and 2006 in the Department of Oncology of the Uludag University School of Medicine, Bursa, Turkey. Membranous or cytoplasmic dominant expression patterns of HER-3 were analyzed using the Rajkumar score and a cytoplasmic 4-point scoring system, respectively. Progression-free survival (PFS) and overall survival (OS) served as the main outcome measures. Results: The median PFS in the study participants was 9 months (interquartile range: 4.5-13 months), whereas the median OS was 20 months (interquartile range: 7.5-28 months). Categorization of the patient population according to HER-3 positive immunohistochemical expression did not reveal any statistically significant difference in terms of both PFS (p=0.70) and OS (p=0.81). The results of multivariable Cox regression analysis indicated that tumor size was the only independent predictor of PFS, whereas estrogen and progesterone receptor status was independently associated with OS. Conclusions: HER-3 immunohistochemical expression did not correlate with outcomes in Turkish patients with metastatic breast cancer. Although our results suggest that HER-3 expression in cancer specimens is not of prognostic significance, further prospective studies are warranted to confirm these results.

Detection and Identification of Mycobacterium Tuberculosis in Patients with Tuberculous Cervical Lymphadenitis by PCR-RFLP (경부 결핵성 임파선염 환자에서 PCR-RELP를 이용한 결핵균의 검출 및 확인)

  • Lee Sang-Sook;Cho Young-Rok;Chun Ji-Min;Choi Yong-Seok;Sohn Eun-Ju;Park Nam-Cho;Park June-Sik
    • Korean Journal of Head & Neck Oncology
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    • v.12 no.2
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    • pp.169-176
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    • 1996
  • Tuberculous cervical lymphadenitis is still an important cause of neck mass in Korea. Tuberculosis is an important differential diagnosis in patients of cervical lymphadenopathy. Rapid and sensitive test for the diagnosis of tuberculosis is essential for the approapiate treatment. Up to now, conventional diagnostic methods for M. tuberculosis were acid-fast bacilli(AFB) stain and culture of M. tuberculosis. The direct microscopic examination of AFB by Ziehl-Neelsen stain is rapid, but often negative. The culture for M. tuberculosis is time-consuming, taking 4 to 8 weeks. Recently various methods to detect Mycobacterial DNA, including PCR and restriction fragment length polymorphism(RFLP) analysis have been reported. Here we represent a simple method for the confirmation of M. tuberculosis and exclusion of the other Mycobacterial species by RFLP analysis and silver staining of polyacrylamide gel electrophoresis after nested PCR for a repetitive DNA sequence(IS986) specific for M. tuberculosis from fresh or paraffin-embedded biopsy specimens. This result leads us to conclude that this method is simple, rapid and possibly applicable to confirm M. tuberculosis and rule out the other Mycobacteria species from the clinical specimens in the clinical laboratories.

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Light and Scanning Electron Microscopic Study on the Structure of the Gastrointestinal Mucosa of the Mole, Talpa micrura coreana (두더지 (Talpa micrura coreana) 위장관 점막의 구조에 대한 광학 및 주사전자현미경적 연구)

  • Kwun, H.S.;Chung, J.W.;Chun, M.H.;Kim, J.;Cha, J.H.;Maeng, S.H.
    • Applied Microscopy
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    • v.15 no.2
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    • pp.111-129
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    • 1985
  • The morphology and histology of the gastrointestinal mucosa of the mole, Talpa micrura coreana (Thomas), were studied using light and scanning electron microscopes. Tissue specimens were taken from body and pyloric portions of the stomach, and from the initial, proximal, middle, distal and terminal portions of the intestine. For light microscopy, tissue blocks were fixed in 10% buffered neutral formalin, embedded in paraffin wax, and sectioned at a thickness of $5{\mu}m$. These sections were stained with hematoxylin-eosin. For scanning electron microscopy, tissue blocks were fixed in 1% glutaraldehyde-1.5% paraformaldehyde, and postfixed in 1% osmium tetroxide, dehydrated in graded alcohol, transferred to isoamylacetate and dried by the critical point drier(Polaron E 3000). Subsequently, specimens were coated with gold and observed with a JSM-35C scanning electron microscope. The results were as follows: The mucous membrane of the body portion of the stomach had numerous irregular folds and the pyloric mucosa formed the strawberry-shaped folds, and general histological structures of each portion were similar to those of man. The intestine could not be differentiated macroscopically and microscopically into small and large intestines. There was no cecum, appendix, taenia coli, haustra coli or appendices epiploicae. In the initial portion (4 mm long), conical or tongue-shaped villi with the height of $143.3{\pm}10.7{\mu}m$ were present, and large mucous glands were seen in the submucosa. In the proximal, middle and distal portions, wavy folds composed of the epithelium and lamina propria were densely and transversely arranged, and their heights were $440.4{\pm}45.5{\mu}m,\;454.4{\pm}19.9{\mu}m\;and\;205.2{\pm}33.5{\mu}m$, respectively. The mucosa of the terminal portion (3 cm long) formed several longitudinal folds, and the intestinal glands were directly opened on the smooth surface of the folds. Aggregated lymphoid follicles were observed in the major portions of the intestine except the initial and terminal portions. There was no circular or semilunar fold throughout the intestine.

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Pathological Factors Affecting DNA Quality in BRAF, EGFR, and KRAS Gene Molecular Tests (BRAF, EGFR, KRAS 유전자 분자병리검사에서 DNA 품질에 영향을 미치는 병리학적인 인자에 관한 연구)

  • Yun, Hyon-Goo;Kim, Bo-Ra;Lee, Joo-Mi;Song, Eun-Ha;Kim, Dong-Hoon
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.4
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    • pp.381-388
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    • 2020
  • The quality control of pathological specimens is important for accurate molecular pathology testing. This study evaluated that specimen factors affecting the DNA quality during tissue processing and sample types for BRAF, EGFR, and KRAS mutations tests. One thousand seven hundred and seventy-two molecular pathology tests were investigated for the factors influencing the DNA quality, such as sample type, formalin fixation time, and reexamination status. Cytology samples stored in a saline solution had better DNA quality than commercial cytology preservation. Tissue samples fixed in formalin within 24 hours had better DNA quality than the samples fixed over 24 hours. Between the types of samples, fresh tissue samples and tissue samples with a high tumor cell density had relatively better DNA quality than the formalin-fixed paraffin-embedded (FFPE) tissues and cytology specimens. Of real-time PCR, the non-PNA Ct value increased proportionally with samples held for longer than 24 hours in formalin, and that the formalin-fixed time affects the sample DNA quality. In conclusion, the appropriate tumor cellularity and 10% neutral formalin fixation time are the most important factors for maintaining the DNA quality. These factors should be managed properly for an accurate pathological molecular test to ensure optimal DNA quality.

Studies on the Phrenic Nerve Fibers in the dog II. On the Myelinated Fibers of the Intradiaphragmatic Branches (개의 횡격막신경섬유(橫隔膜神經纖維)에 관(關)한 연구(硏究) 제(第)II보(報) 횡격막신경(橫隔膜神經)의 횡격막내분기(橫隔膜內分技)의 유수섬유(有髓纖維)에 관(關)하여)

  • Ko, Kwang Doo;Yoon, Suk Bong;Lee, Joon Sup
    • Korean Journal of Veterinary Research
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    • v.9 no.1
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    • pp.1-9
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    • 1969
  • The experimental studies were performed to observe the characteristics of the myelinated nerve fibers of the tradiaphragmatic branches of phrenic nerves in dog In the work to be reported five mixed breed Korean dog were used, they were one year old and he body eights were about 12 Kgm. in average with healthy conditions. The specimens (nerve) were taken at the point of 1.5 cm posterior part from the left and right phrenic nerves entrance to the diaphragm, and the dorso-lateral and dorso-medial branches of phrenic nerves vere also, taken at the point of 0.5 cm distal part from their branching portion. The specimens were fixed for 24 hrs. in Flemming's solution, and embedded in paraffin. The paraffin section. were cut at 6 microns and stained with Walter's modification of Weigert-pal method for the myelinated fibers. Microphotographs were taken and enlarged into 750 times of the actual size, and the diameter of the photographic images of the myelinated fibers were measured by the scale on the transparent Percepex Plate. The following conclusions were made: 1. The percentage of distribution of the several intradiaphragmatic branches of phrenic nerves were as follows. Ventral branches were 34.97%, lateral branches were 37.90%, dorsal branches were 27.13%, and the dorsal branches were branched again into dorso-lateral branches(54.22%) and dorso-medial branches (45. 78%). 2. The mean value and S.D. of the total numbers of the myelinated nerve fibers at the left ventral branches were $490.80{\pm}12$, and at the right ventral branches were $486.6{\pm}13$. Total average cross sectional area on the left ventral branches were 38,000. $6{\pm}136{\mu}^2$, and the right ventral branches were $37,150.2{\pm}1.782{\mu}^2$. 3. The mean value and S.D. of the total numbers of the myelinated nerve fibers at the left lateral branches were $533.8{\pm}8$, and at the right lateral branches were $525.6{\pm}7$. Total average cross sectional area of the left lateral branches were $41,582{\pm}1,170{\mu}^2$, and the right lateral branches were $40,454.8{\pm}812{\mu}^2$. 4. The mean value and S.D. of the total numbers of the myelinated nerve fibers at the left dorsal branches were $378.2{\pm}14$, and at the right dorsal branches were $380.2{\pm}8$. Total average cross sectional area of the left dorsal branches were $27,771{\pm}1,256{\mu}^2$, and the right dorsal branches were $27,507.2{\pm}645{\mu}^2$. 5. The mean value and S.D. of the total numbers of the myelinated fibers at the left dorso-lateral branches were $205.4{\pm}15$, and at the right dorso-lateral branches were $210.6{\pm}17$. Total average cross sectional area of the left dorso lateral branches were $15,354. 8{\pm}1,519{\mu}^2$, and the right dorsal branches were $15,887{\pm}1,297{\mu}^2$. 6. The mean value and S.D. of the total numbers of the myelinated nerve fibers at the left dorso-medial branches were $175{\pm}14$, and at the right dorso-medial branches were $176.2{\pm}17$. Total average cross sectional area of the left dorso-medial branches were $13,037.4{\pm}944{\mu}^2$, and at the right dorso-medial branches were, $13,103{\pm}1,373{\mu}^2$. 7. The highest frequent distribution of the intradiaphragmatic branches of phrenic nerves were found in the $10-12{\mu}^2$ groups, and they were almost 30% of the total groups. 8. The largest fibers diameter were in the $14-16{\mu}^2$ groups, and these were shown the lowest frequent distribution. 9. The largest cross sectional area of the intradiaphragmatic branches of phrenic nerves were found in the $10-12{\mu}^2$ groups. 10. All of the intradiaphragmatic branches of phrenic nerves were unimodal which has a peak in the $10-12{\mu}^2$ groups.

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The Clinical Significance of Cathepsin D and p53 Expression in Locally Advanced Rectal Cancer (국소진행된 직장암에서 Cathepsin D와 p53 발현의 임상적 의의)

  • Kim, Jun-Sang;Lee, Sheng-Jin;Kim, Jin-Man;Cho, Moon-June
    • Radiation Oncology Journal
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    • v.26 no.1
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    • pp.56-64
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    • 2008
  • Purpose: Cathepsin D(CD) is a lysosomal acid proteinase that is related to malignant progression, invasion, and a poor prognosis in several tumors. The aim of this study was to evaluate the prognostic clinical significance of CD and p53 expression in pretreatment biopsy specimens from patients with locally advanced rectal cancer who were treated with preoperative chemoradiation. Materials and Methods: Eighty-nine patients with locally advanced rectal cancer(cT3/T4 or N+) were included in this study. Preoperative chemoradiation consisted of a dose of 50.4 Gy of pelvic radiation and two concurrent cycles of administration of 5-fluorouracil and leucovorin. Surgery was performed six weeks after chemoradiation. CD and p53 expression in pretreatment formalin-fixed paraffin-embedded tumor biopsy specimens were assessed by immunohistochemical staining using a CD and p53 monoclonal antibodies. The threshold value for a positive stain in tumor tissue and stromal cells was 1+ intensity in 10% of the tumors or stromal cells, respectively. Results: Positive CD expression was found in 57(64%) of the tumors and 32(35%) of the stromal cell specimens. There was no association with CD expression of the tumor or stromal cells and patient characteristics. There was a correlation between tumor CD expression with stromal cell CD expression(p=0.01). Overexpression of p53 was not a significant prognostic factor. The 5-year overall survival(OS) and disease-free survival(DFS) rates were not different between tumor CD-negative and positive patient biopsy samples(69% vs. 65%, 60% vs. 61%, respectively). The 5-year OS rates in the tumor-negative/stromal cell-negative, tumor-negative/stromal cell-positive, tumor-positive/stromal cell-negative and tumor-positive/stromal cell-positive biopsy samples were 75%, 28%, 62%, and 73%, respectively. Stromal cell staining only without positive tumor staining demonstrated the worst overall survival prognosis for patients(p=0.013). Conclusion: Overexpression of p53 in rectal biopy tissue was not associated with prognostic significance. In the pretreatment biopsy specimens, an exclusive increase in CD expression in stromal cells without tumor expression was related to poor overall survival in patients with locally advanced rectal cancer treated with preoperative chemoradiation.

Claritromycin Resistance and Helicobacter pylori Genotypes in Italy

  • Francesco Vincenzo De;Margiotta Marcella;Zullo Angelo;Hassan Cesare;Valle Nicolar Della;Burattini Osvaldo;D'Angel Roberto;Stoppino Giuseppe;Cea Ugo;Giorgio Floriana;Monno Rosa;Morini Sergio;Panella Carmine
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.660-664
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    • 2006
  • The relationship between H. pylori clarithromycin resistance and genetic pattern distribution has been differently explained from different geographic areas. Therefore, we aimed to assess the clarithromycin resistance rate, to evaluate the bacterial genetic pattern, and to search for a possible association between clarithromycin resistance and cagA or vacA genes. This prospective study enrolled 62 consecutive H. pylori infected patients. The infection was established by histology and rapid urease test. Clarithromycin resistance, cagA and vacA status, including s/m subtypes, were assessed on paraffin-embedded antral biopsy specimens by TaqMan real time polymerase chain reaction (PCR). Primary clarithromycin resistance was detected in 24.1 % of cases. The prevalence of cagA was 69.3%, and a single vacA mosaicism was observed in 95.1 % cases. In detail, the s1m1 was observed in 23 (38.9%) patients, the s1m2 in 22 (37.2%), and the s2m2 in 14 (23.7%), whereas the s2m1 combination was never found. The prevalence of cagA and the vacA alleles distribution did not significantly differ between susceptible and resistant strains. Primary clarithromycin resistance is high in our area. The s1m1 and s1m2 are the most frequent vacA mosaicisms. There is no a relationship between clarithromycin resistance and bacterial genotypic pattern and/or cagA positivity.

Epidermal Growth Factor Receptor Mutations in Non-Small Cell Lung Cancers in a Multiethnic Malaysian Patient Population

  • Liam, Chong-Kin;Leow, Hwong-Ruey;How, Soon-Hin;Pang, Yong-Kek;Chua, Keong-Tiong;Lim, Boon-Khaw;Lai, Nai-Lang;Kuan, Yeh-Chunn;Pailoor, Jayalakshmi;Rajadurai, Pathmanathan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.321-326
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    • 2014
  • Background: Mutations in the tyrosine kinase domain of the epidermal growth factor receptor (EGFR) in non-small cell lung cancer (NSCLC) are predictive of response to EGFR-targeted therapy in advanced stages of disease. This study aimed to determine the frequency of EGFR mutations in NSCLCs and to correlate their presence with clinical characteristics in multiethnic Malaysian patients. Materials and Methods: In this prospective study, EGFR mutations in exons 18, 19, 20 and 21 in formalin-fixed paraffin-embedded biopsy specimens of consecutive NSCLC patients were asessed by real-time polymerase chain reaction. Results: EGFR mutations were detected in NSCLCs from 55 (36.4%) of a total of 151 patients, being significantly more common in females (62.5%) than in males (17.2%) [odds ratio (OR), 8.00; 95% confidence interval (CI), 3.77-16.98; p<0.001] and in never smokers (62.5%) than in ever smokers (12.7%) (OR, 11.50; 95%CI, 5.08-26.03; p<0.001). Mutations were more common in adenocarcinoma (39.4%) compared to non-adenocarcinoma NSCLCs (15.8%) (p=0.072). The mutation rates in patients of different ethnicities were not significantly different (p=0.08). Never smoking status was the only clinical feature that independently predicted the presence of EGFR mutations (adjusted OR, 5.94; 95%CI, 1.94-18.17; p=0.002). Conclusions: In Malaysian patients with NSCLC, the EGFR mutation rate was similar to that in other Asian populations. EGFR mutations were significantly more common in female patients and in never smokers. Never smoking status was the only independent predictor for the presence of EGFR mutations.

Expression of ERCC1, MSH2 and PARP1 in Non-small Cell Lung Cancer and Prognostic Value in Patients Treated with Platinum-based Chemotherapy

  • Xie, Ke-Jie;He, Hong-Er;Sun, Ai-Jing;Liu, Xi-Bo;Sun, Li-Ping;Dong, Xue-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2591-2596
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    • 2014
  • Purpose: To evaluate the prognostic value of the expression of excision repair cross-complementation group l (ERCC1), MutS protein homolog 2 (MSH2) and poly ADP-ribose polymerase 1 (PARP1) in non-small-cell lung cancer patients receiving platinum-based postoperative adjuvant chemotherapy. Methods: Immunohistochemistry was applied to detect the expression of ERCC1, MSH2 and PARP1 in 111 cases of non-small cell lung cancer paraffin embedded surgical specimens. Through og-rank survival analysis, we evaluated the prognostic value of the ERCC1, MSH2, PARP1 and the related clinicopathological factors. COX regression analysis was used to determine whether ERCC1, MSH2 and PARP1 were independent prognostic factors. Results: In the enrolled 111 non-small cell lung cancer patients, the positive expression rate of ERCC1, MSH2 and RARP1 was 33.3%, 36.9% and 55.9%, respectively. ERCC1 (P<0.001) and PARP1 (P=0.033) were found to be correlated with the survival time while there was no correlation for MSH2 (P=0.298). Patients with both ERCC1 and PARP1 negative cancer had significantly longer survival time than those with ERCC1 (P=0.042) or PARP1 (P=0.027) positive alone. Similalry, the survival time of patients with both ERCC1 and PARP1 positive cancer was shorter than those with ERCC1 (P=0.048) or PARP1 (P=0.01) positive alone. Conclusion: Patients with ERCC1 or PARP1 negative non-small cell lung cancer appear to benefit from platinum-based postoperative adjuvant chemotherapy.

Prevalence of Aflatoxin Induced p53 Mutation at Codon 249 (R249s) in Hepatocellular Carcinoma Patients with and without Hepatitis B Surface Antigen (HBsAg)

  • Chittmittrapap, Salyavit;Chieochansin, Thaweesak;Chaiteerakij, Roongruedee;Treeprasertsuk, Sombat;Klaikaew, Naruemon;Tangkijvanich, Pisit;Komolmit, Piyawat;Poovorawan, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7675-7679
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    • 2013
  • Background: A missense mutation in exon 7 (R249S) of the p53 tumor suppressor gene is characteristic of aflatoxin B1 (AFB1) exposure. AFB1 is believed to have a synergistic effect on hepatitis virus B (HBV) carcinogenesis. However, results of studies comparing R249S prevalence among patients are conflicting. The aim of this study was to determine the prevalence of the R249S mutation in hepatocellular carcinoma (HCC) patients with or without positive HBsAg. Materials and Methods: Paraffin embedded liver tissues were obtained from 124 HCC patients who underwent liver resection and liver biopsy in King Chulalongkorn Memorial Hospital. Restriction fragment length polymorphism (RFLP) was utilized to detect the R249S mutation. Positive results were confirmed by direct sequencing. Results: Sixty four (52%) patients were positive for HBsAg and 18 (15%) were anti-HCV positive. 12 specimens tested positive by RFLP. Ten HCC patients (8.1%) were confirmed to be R249S positive by Sanger sequencing (AGG to AGT). Out of these 10, six were HBsAg positive, and out of the remaining 4, two were anti-HCV positive. The R249S prevalence among HCC patients with positive HBsAg was 9.4% compared to 6.7% for HBsAg negative samples. Patients with the R249S mutation were younger ($55{\pm}10$ vs $60{\pm}13$ year-old) and tended to have a more advanced Edmonson-Steiner grade of HCC, although differences did not reach statistical significance. Conclusions: Our study shows moderate prevalence of aflatoxin B1-related p53 mutation (R249S) in HCC with or without HBsAg. HBsAg positive status was not associated with R249S prevalence.