Clinical significance of miR-140-5p and miR-193b expression in patients with breast cancer and relationship to IGFBP5.

The functional role of IGFBP5 in breast cancer is complicated. Experimental and bioinformatics studies have shown that IGFBP5 is targeted by miR-140-5p and miR-193b, although this has not yet been proven in clinical samples. The aim of this study was to evaluate the expression of miR-140-5p and miR-193b in breast cancer and adjacent normal tissue and assess its correlation with IGFBP5 and the clinicopathological characteristics of the tumors. IGFBP5 protein expression was analyzed immunohistochemically and IGFBP5, miR-140 and miR-193b mRNA expression levels were analyzed with real-time RT-PCR. Tumor tissue had higher miR-140-5p expression than adjacent normal tissue (p = 0.015). Samples with no immunohistochemical staining for IGFBP5 showed increased miR-140-5p expression (p = 0.009). miR-140-5p expression was elevated in invasive ductal carcinomas (p = 0.002), whereas basal-like tumors had decreased expression of miR-140-5p compared to other tumors (p = 0.008). Lymph node-positive samples showed an approximately 13-fold increase in miR-140-5p expression compared to lymph node-negative tissue (p = 0.049). These findings suggest that miR-140-5p, but not miR-193b, could be an important determinant of IGFBP5 expression and clinical phenotype in breast cancer patients. Further studies are needed to clarify the expressional regulation of IGFBP5 by miR-140-5p.

A rare cause of mechanical intestinal obstruction: Pharmacobezoar.

A bezoar is a hard, and solid, foreign body located in the gastrointestinal tract that may recur. Bezoar is classified according to its origin. Pharmacobezoars develop in the gastrointestinal tract due to alterations in anatomical structure and/or intestinal motility. In this paper, a case, not yet defined in the literature, of a pharmacobezoar causing a mechanical obstruction that is accompanied by a malignancy in the colon is reported, with the aim of contributing to the literature.

MicroRNA-21 as an indicator of aggressive phenotype in breast cancer.

OBJECTIVE: The recently discovered microRNAs (miRNAs) are non-protein-coding, endogenous small RNAs. The aim of this study was to investigate the relationship between microRNA-21 (miR-21) and the clinicopathological features of breast cancer. MATERIALS AND METHODS: Formalin-fixed, paraffin-embedded (FFPE) tissue samples were collected from 15 patients who had undergone surgery for primary breast cancer. The miR-21 expression levels in normal and cancer tissues were analyzed using real-time quantitative reverse transcriptase polymerase chain reaction (real-time qRT-PCR). The correlations between the miR-21 expression level and the stage of disease, the tumor size, lymph node involvement, hormone receptor status, human epidermal growth factor receptor 2 (HER2) status, and disease-free survival (DFS) were investigated. RESULTS: The miR-21 expression levels were significantly higher in patients with stage III disease, patients with more than 3 axillary lymph node metastases and HER2-positive patients than in patients with stage I-II disease, patients with 1-3 axillary lymph node metastases and HER2-negative patients (p = 0.005, p = 0.037, and p = 0.014, respectively). Patients with high miR-21 expression level had a significantly shorter DFS than patients with low miR-21 expression level (median DFS: 18 and 56 months, respectively; p = 0.004). CONCLUSION: These results show that miR-21 is an indicator of an aggressive breast cancer phenotype and that it may be a new therapeutic target in the treatment of breast cancer in the future.

Role of gamma probe-assisted intraoperative sentinel lymph node evaluation in predicting axillary breast cancer metastasis after neoadjuvant chemotherapy.

OBJECTIVE: Intraoperative sentinel lymph node biopsy is a universally accepted technique to identify patients who are candidates for axillary lymph node dissection during breast cancer surgery. However, there is controversy over its use in patients who underwent preoperative neoadjuvant chemotherapy. This study aimed to examine the diagnostic value of gamma probe-assisted intraoperative sentinel lymph node examination with frozen section in breast cancer patients who had undergone preoperative neoadjuvant chemotherapy. METHODS: This retrospective study included 94 tumors diagnosed with stage IIA, IIB or IIIA invasive breast cancer with locoregional lymph node metastasis who underwent surgical treatment after neoadjuvant chemotherapy. Intraoperatively, axillary sentinel lymph node sampling was done using radioactive colloid and gamma probe and materials were examined with frozen section method. Patients with positive sentinel nodes underwent axillary resection. Histopathological examination of all surgical samples was done postoperatively. RESULTS: In 87 of 94 tumors (92.6%), a sentinel lymph node could be identified using the method. The sensitivity, specificity and accuracy of the method for predicting axillary macro metastasis were 85.7, 86.5 and 86.2%, respectively, with 5.7% false negative rate. CONCLUSIONS: Sentinel lymph node identification using preoperative scintigraphy and intraoperative use of gamma probe seems to be a feasible and efficient method in terms of differentiating patients that require axillary lymph node dissection during breast cancer surgery, even when they have received neoadjuvant chemotherapy. Further large prospective studies allowing subgroup analyses are warranted.

Prognostic Significance of Tumor Tissue NeuGcGM3 Ganglioside Expression in Patients Receiving Racotumomab Immunotherapy.

Expression of N-glycolyl GM3 (NeuGcGM3) ganglioside was detected in the tumor specimens of patients who were on Racotumomab anti-idiotype vaccine maintenance treatment, and prognostic significance as a biomarker was investigated. No statistically significant association was observed in the multivariate analysis between overall survival and tissue NeuGcGM3 IHC levels. Although numerically there was a difference favoring less intense IHC for better prognosis, this did not reach statistical power. However, there was a strong correlation between Racotumomab doses and overall survival (OS). Mean OS of the patient with more than 10 Racotumomab application was significantly longer than the patient who had less than 10 injections (70.7 months vs. 31.1 months, p < 0.001). We propose that, regardless of staining intensity, the presence of NeuGcGM3 in patient tissues might be an indicator of benefit in Racotumomab treatment.

The significance of histopathologic patterns in positive tuberculin skin test site.

BACKGROUND: The histologic features of tuberculin skin test site is not uniform. It may be related to status of tuberculosis. METHODS: Forty-eight purified protein derivative (PPD) positive-cases were chosen for the study. Thirty of the subjects had active tuberculosis. As previously reported, the histologic pattern of inflammatory reaction seen in the test site classified into three type: (a) Perivascular (PV)-type, (b) Basal spongiotic dermatitis (BSD)-type and (c) Erythema multiforme (EM)-type. The frequencies of histological patterns in active tuberculosis and latent tuberculosis were statistically analyzed. RESULTS: In active tuberculosis group including 30 patient, 17 (56.7%) EM-type, 9 (30%) BSD-type and 4(13.3%) PV-type inflammation were seen. Among 18 latent tuberculosis, there were 2 (11.1%) EM-type, 7 (38 8%) BSD-type and 9 (50%) PV-type inflammatory reactions. The EM-type inflammation was more common in active tuberculosis group. Bulla formation was seen in seven subjects with active tuberculosis. CONCLUSION: The histological pattern of PPD reaction site may be an important sign reflecting the nature of the tuberculosis, which may be either latent or active. The bulla formation is an important sign for active pulmonary tuberculosis. Further, detailed immunohistopathologic studies of PPD reaction with large number of cases may give important clues about tuberculosis immunology.

Histologic evidence of new collagen formation using a Q-switched Nd:YAG laser in periorbital rhytids.

OBJECTIVES: Non-ablative laser treatment has been used for improving dermal toning. Laser application to dermis causes new collagen formation in terms of wound healing. We aimed to study mainly histological changes in skin after the use of a Q-switched Nd:YAG laser in non-ablative treatment of wrinkles. METHODS: The laser was adjusted to a fluency of 7 J/cm2 with a spot size of 3 mm and a pulse rate of 10 Hz to treat periorbital wrinkled areas. None of the patients had received filling materials, botulinum toxin injections or any dermabrasion procedures. All laser sessions were held every 15 days for a total of six sessions and all patients were photographed before treatment and then 2 months after the last treatment. Histological examinations were performed before laser treatment and 1 month after the final treatment. RESULTS: Four of eight individuals showed clinical improvement. The histological proportion of collagen fibers was measured by using the Samba method. An increase in the mean optical densities (MOD) of collagen fibers compared with baselines was statistically significant in all patients (p<0.05). CONCLUSION: The 1064-nm Q-switched Nd:YAG lasers appear to be safe and efficient for non-ablative remodeling of periorbital wrinkles.

Plexin C1 Marks Liver Cancer Cells with Epithelial Phenotype and Is Overexpressed in Hepatocellular Carcinoma.

Background and Aims: Hepatocellular carcinoma is an aggressive malignancy of the liver and is ranked as the sixth most common cancer worldwide. There is still room for novel markers to improve the diagnosis and monitoring of HCC. Our observations in cancer databases that PLXNC1 is upregulated in HCC led us to investigate the expression profile of Plexin C1 mRNA and protein in HCC cell lines and tissues. Methods: A recombinant protein encompassing part of the extracellular domain of Plexin C1 was used as an antigen for monoclonal antibody development. Transcript and protein levels of Plexin C1 in HCC cell lines were determined by RT-qPCR and Western blotting, respectively. In vivo evaluation of Plexin C1 expression in HCC tissues was accomplished by immunohistochemistry studies in tissue microarrays. Results: A monoclonal antibody, clone PE4, specific to Plexin C1, was generated. In silico and in vitro analyses revealed a Plexin C1-based clustering of well-differentiated HCC cell lines. Staining of HCC and nontumoral liver tissues with PE4 showed a membrane-localized overexpression of Plexin C1 in tumors (p=0.0118). In addition, this expression was correlated with the histological grades of HCC cases. Conclusions: Plexin C1 distinguishes HCC cells of epithelial characteristics from those with the mesenchymal phenotype. Compared to the nontumoral liver, HCC tissues significantly overexpress Plexin C1. The newly generated PE4 antibody can be evaluated in larger HCC cohorts and might be exploited for the examination of Plexin C1 expression pattern in other epithelial malignancies.

Antioxidants vitamin E and C attenuate hepatic fibrosis in biliary-obstructed rats.

AIM: To investigate whether antioxidants vitamin E and C can retard development of hepatic fibrosis in the biliary-obstructed rats. METHODS: Fifty Wistar albino rats were randomly assigned to 5 groups (10 rats in each). Bile duct was ligated in 40 rats and they were treated as follows: group vitC, vitamin C 10 mg/kg sc daily; group vitE, vitamin E 15 mg/kg sc daily; group vitEC, both of the vitamins; bile duct-ligated (BDL, control) group, physiological saline sc. The fifth group was assigned to sham operation. At the end of fourth week, the rats were decapitated, and hepatic tissue biochemical collagen content and collagen surface area were measured. Hepatic tissue specimens were histopathologically evaluated according to Scheuer system. Serum hyaluronate levels were measured by ELISA method. RESULTS: Despite being higher than sham group, hepatic collagen level was significantly decreased in each of the vitC, vitE and vitEC groups (32.7 +/- 1.2, 33.8 +/- 2.9, 36.7 +/- 0.5 mug collagen/mg protein, respectively) compared to BDL (48.3 +/- 0.6 mg collagen/g protein) (P < 0.001 for each vitamin group). Each isolated vitamin C, isolated vitamin E and combined vitamin E/C supplementation prevented the increase in hepatic collagen surface density (7.0% +/- 1.1%, 6.2% +/- 1.7%, 12.3% +/- 2.0%, respectively) compared to BDL (17.4% +/- 5.6%) (P < 0.05 for each). The same beneficial effect of vitamin C, vitamin E and combined vitamin E/C treatment was also observed on the decrease of serum hyaluronate levels compared to BDL group (P < 0.001). The relative liver and spleen weights, serum transaminases, cholestatic enzymes, bilirubins and histopathological inflammation scores were not different between the antioxidant treatment groups and the control. However, fibrosis staging scores were obviously reduced only in the vitamin E/C combination group (vit EC: 2.4 +/- 0.8 vs BDL: 3.1 +/- 0.7; P < 0.05). CONCLUSION: Each antioxidant vitamin E, vitamin C and their combination retard hepatic fibrosis in biliary-obstructed rats. Oxidative stress may play a role in the pathogenesis of hepatic fibrosis in secondary biliary cirrhosis.

Histologic Evidence of New Collagen Formulation Using Platelet Rich Plasma in Skin Rejuvenation: A Prospective Controlled Clinical Study.

BACKGROUND: Platelet-rich plasma (PRP) is an autologous concentration of human platelets contained in a small volume of plasma and has recently been shown to accelerate rejuvenate aging skin by various growth factors and cell adhesion molecules. OBJECTIVE: This study was conducted to evaluate the efficacy and safety of intradermal injection of PRP in the human facial rejuvenation. METHODS: This study was a prospective, single-center, single-dose, open-label, non-randomized controlled clinical study. PRP injected to the upper site of this right infra-auricular area and all face. Saline was injected to the left infra-auricular area. Histopathological examinations were performed before PRP treatment, 28 days after the PRP, and saline (control) treatments. RESULTS: Twenty women ranging in age from 40 to 49 years (mean age, 43.65±2.43 years) were enrolled in the study. The mean optical densities (MODs) of collagen in the pre-treatment, control, and PRP-treated area were measured. They were 539±93.2, 787±134.15, 1,019±178, respectively. In the MOD of PRP, 89.05 percent improvement was found when MOD of PRP was compared with MOD of pre-treatment. The mean MOD of collagen fibers was clearly highest on the PRP side (p<0.001). The PRP-to-saline improvement ratio (89.05% to 46.01%) was 1.93:1. No serious side effects were detected. CONCLUSION: PRP increases dermal collagen levels not only by growth factors, but also by skin needling (the mesotherapy technique 'point by point'). PRP application could be considered as an effective (even a single application) and safety procedure for facial skin rejuvenation.

Dental technician's pneumoconiosis: mineralogical analysis of two cases.

Pneumoconiosis was diagnosed by open lung biopsy in two dental technicians who had interstitial lung disease. Mineralogical analysis was performed to investigate the origin of the dust that had been inhaled. A marked accumulation of silicon and phosphorus was found in both cases. The hard metals chromium and cobalt were also found. Dental technician's pneumoconiosis is a complex pneumoconiosis in which such dust and hard metals may play a role.

Should sentinel lymph node mapping be performed for colorectal cancer?

BACKGROUND/AIMS: Correct determination of lymphatic nodal statement is essential to stage correctly and to predict survival. As it is vital to make an assessment about the adjacent lymph node(s), this study was designed to compose a sensitive detection on the sentinel lymph nodes (SLN) indicating tumoral lymphatic basin using advanced pathologic examination. MATERIALS AND METHODS: From June 2002 to June 2003, this prospective study was performed in 41 patients undergoing standard resection for colorectal cancer. In this study we employed the ex-vivo SLN mapping technique. RESULTS: At least one SLN in 37 of 41 patients was identified (90.2%). The lymph nodes (LN) from those patients were studied by hematoxylin and eosin dye (H&E) and multisectioning. Twenty of 37 patients with trace of the metastasis were found. The remaining 17 patients without any metastatic LN by H&E underwent clarification of micrometastases (MM) using immunohistochemical (IHC) staining technique. Two patients (11.7%) had MM in the SLN(s). Upstaging was evaluated in those two. The sensitivity of SLNs was obtained as 90%. Two patients with no metastatic SLN had metastasis in the non-sentinel LNs. CONCLUSIONS: In the LNs from the basin of tumor, MM exposed by IHC staining was still not obvious to indicate poor prognosis. The need for treatment adjustment in those patients is clear since the upstaging was evident.

EGFR and KRAS mutations in Turkish non-small cell lung cancer patients: a pilot study.

EGFR and KRAS mutation profile in non-small cell lung cancers (NSCLCs) shows wide variations due to geographic and ethnic background. We aimed to determine the frequency and types of EGFR and KRAS mutations in a sample group of Turkish NSCLC cases. The study included 14 adenocarcinomas (ACs), 11 squamous cell carcinoma (SCC) patients selected from archival material including small biopsy or surgical specimens. Their formalin fixed paraffin-embedded tumor tissues were used for genomic DNA extraction for EGFR exon 19 and 21, and KRAS exon 2 mutations. Eleven NSCLCs (44 %) had EGFR mutations. Exon 19 and 21 mutations were found in 8 (32 %) and 5 (20 %) cases. Two cases showed double EGFR mutations. In ACs, 5 (35.7 %) patients had EGFR gene mutation, 3 in exon 19 and 3 in exon 21. In SCCs, 6 (54.5 %) cases had EGFR mutation, 5 in exon 19 and 2 in exon 21. All exon 19 mutations were deletion-type mutations. For exon 21, 3 cases had L858R point mutation (CTG>CGG) and two cases showed deletion-type mutations. Six (24 %) NSCLCs showed KRAS mutations (three ACC, three SCC), 5 codon 12 mutations (G>T, T>C, G>A) and one codon 13 mutation (G>T). Three NSCLC cases showed both EGFR and KRAS mutations together. The profile of KRAS mutation in our AC cases was quite similar to those seen in the Western countries; however, frequency and clustering of EGFR mutations were similar to those seen in the Eastern countries.

Stromal cells do not share KRAS mutations that the epithelial component harbors in colorectal carcinomas.

Epithelia and stroma are the main components of carcinomas that have impact on carcinogenesis. Many of the genetic changes harboring in the epithelia may possibly be seen in stromal cells during neoplastic transformation. We intended to investigate weather KRAS mutations are shared by the stromal cells in colorectal adenocarcinomas. Forty cases with KRAS mutation were studied. Glandular/epithelial and the stromal components of each primary tumor were collected and KRAS mutation analysis was performed. None of the cases revealed KRAS mutations in stromal integral. We concluded that stromal cells in colorectal carcinoma do not share KRAS mutations that the epithelial component harbors.

[A new remedy in pathology practice: molecular solution to sample mix-up]. / Patoloji Pratiginde Yeni Bir Çare Uygulamasi: Örnek Karisikligina Moleküler Çözüm.

OBJECTIVE: Although very rare, suspicious situations about the identity of diagnostic tissue material have been encountering in pathology practice. Such situations undoubtedly have the potential to create undesirable results. In the present study, an application targeting getting rid of any doubts about the identity of the diagnostic tissue samples is described. MATERIAL AND METHOD: A combination of short tandem repeats (STR) of the human genome consisting of CSF1PO, TH01, TPOX, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539 and Penta E were selected on the basis of ease of application and bioinformatic discrimination power. Possible forms of diagnostic tissue mix up were set in 3 different models with 3 diagnostic tissue samples of 2 different cases. Of the tissue samples selected, A (salivary gland) and B (striated muscle) belonged to the same case and C (uterus wall) belonged to another case. In the first model, there was no problem about tissue identity (M1: A/B). In the second model, two different diagnostic material were mixed up (M2: B/C). In the last model, there were 3 diagnostic material obtained from 2 different cases (M3: A/B/C). DNA was extracted from all tissue samples and all of the selected 10 STR were amplified with specially designed primers by PCR. After chemical denaturation, amplicons were submitted to polyacrylamide gel electrophoresis for discrimination of single DNA strands according to their conformation polymorphism (SSCP). Special patterns of each STR in the gel matrix obtained from M1, M2 and M3 models, were evaluated on the principle of being 'same or different' to determine the diagnostic material identity. RESULTS: Each of the salivary gland, striated muscle and uterus wall samples were correctly identified (matched with the right source cases) after evaluating 10 different STR SSCP patterns designed under M1, M2 and M3 models. CONCLUSION: This application targeting to solve diagnostic tissue identity problems is a simple and cheap application of SSCP and its efficacy was proven on the designed models.

A valuable adjunct to FNA diagnosis of papillary thyroid carcinoma: in-house PCR assay for BRAF T1799A (V600E).

The diagnostic approach to thyroid nodules generally starts with FNA cytology. However, approximately one-fifth of cytologic evaluations yield indeterminate cytological findings but only 20% of cases with indeterminate thyroid nodule cytology have a cancer diagnosis, emphasizing the need for an effective ancillary test based on FNA material to help prevent unnecessary surgery. Detection of BRAFV600E mutation, the genetic signature of papillary thyroid carcinoma (PTC) in FNA material provides an invaluable diagnostic adjunct to overcome the limitations of FNA cytology. There are many ways to detect V600E, such as direct DNA sequencing, allele-specific PCR and hybridization-based colorimetric methods. In this study, a newer simple PCR method is presented that removes requirements for sequencing special equipment and commercial kits. Two forward primers including the mutant sequence specific (F2), and one common reverse (R) primer were optimized to generate a 241 bp fragment (F1R), an internal PCR control, and a 141 bp fragment (F2R) denoting the presence of V600E. Sensitivity studies revealed that the assay is capable of detecting V600E even in 1 ng of DNA. Direct sequencing data of 241 bp F1R fragment proved the specificity of the assay. For validation studies of the sequence specific multiplex PCR assay, archival FNA slides were used in a group of thyroid lesions including PTC, follicular carcinoma, follicular adenoma, Hashimoto thyroiditis, and benign thyroid nodules. The newer PCR-based method presented in this study is a practical, inexpensive one-step assay to detect the BRAF T1796A mutation on FNA samples.