[Show abstract][Hide abstract] ABSTRACT: To develop a novel polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) protocol using a new genomic marker sequence and a novel set of restriction enzymes in order to detect and discriminate 2 Echinococcus species, E. granulosus and E. multilocularis, found in formalin-fixed paraffin-embedded (FFPE) human tissues.
DNA was isolated from 11 FFPE human tissue samples positive for cystic echinococcosis or alveolar echinococcosis. A mitochondrial genomic marker region was amplified and sequenced using a novel primer pair and a new PCR-RFLP protocol was developed for the detection and discrimination of E. granulosus and E. multilocularis using a set of restriction enzymes including AccI, MboI, MboII, and TsoI.
The selected marker region was amplified using DNA isolated from FFPE human tissue samples positive for cystic echinococcosis or alveolar echinococcosis and the discrimination of E. granulosus and E. multilocularis was accomplished by use of the novel PCR-RFLP method.
In this PCR-RFLP protocol, use of any single restriction enzyme is enough for the discrimination of E. granulosus and E. multilocularis. The PCR-RFLP protocol can be potentially used for the discrimination of 5 other Echinococcus species: E. oligarthus, E. shiquicus, E. ortleppi, E. canadensis, and E. vogeli.
Full-text · Article · Dec 2014 · Turkish Journal of Medical Sciences
[Show abstract][Hide abstract] ABSTRACT: Epidermal growth factor receptor (EGFR) and its ligands are commonly expressed by synovial cells. The aim of the present study was to detect the potential effect of lapatinib an inhibitor of EGFR tyrosine kinases on collagen-induced arthritis. Thirty Wistar albino female rats were randomized into three groups. Arthritis was induced by intradermal injection of chicken type II collagen with incomplete Freund's adjuvant. Serum TNF-α, IL-17, and malondialdehyde (MDA) levels were analyzed. Tissue superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) activities, and nuclear factor erythroid 2-related factor-2 (Nrf2) and heme oxgenase-1 (HO-1) expressions were determined. TNF-α, IL-17 and MDA levels, and Nrf2 and HO-1 expressions were lower in lapatinib-treated (30 mg/kg/day) group compared to sham group, while SOD, catalase, and GPx activities were higher (p < 0.05). Moreover, lapatinib ameliorated perisynovial inflammation and cartilage-bone destruction (p < 0.001). In conclusion, EGFR may have prominent pathogenic role and lapatinib may be an effective therapeutic option for arthritis.
[Show abstract][Hide abstract] ABSTRACT: Pemetrexed (PMTX) is an anti-folate drug as methotrexate. The purpose of this study was to assess the efficacy of PMTX on collagen-induced arthritis (CIA). Forty Wistar albino rats were randomized into four groups. Arthritis was induced by intradermal injection of chicken type II collagen combined with incomplete Freund's adjuvant. Animals were sacrificed at the 15th day after the onset of arthritis. Tumor necrosis factor alpha (TNF-α), interleukin (IL)-17, and malondialdehyde (MDA) levels were increased, and superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities and the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were decreased in the arthritis group. In the PMTX-treated (0.2 and 1 mg/kg/week i.p.) groups, the levels of TNF-α, IL-17, and MDA were decreased; the activities of SOD, CAT, and GPx and the expressions of Nrf2 and HO-1 were restored, and perisynovial inflammation and cartilage-bone destruction were decreased. PMTX has anti-arthritic potential in the CIA model and may be a therapeutic agent for rheumatoid arthritis.
[Show abstract][Hide abstract] ABSTRACT: Although, the exact mechanisms underlying the development of the metabolic syndrome (MetS) are not still completely understood, obesity, circulated peptide hormone levels and their interaction with genetic factors are considered largely responsible. The purpose of this study is to explore how the levels of ghrelin, obestatin (OBS) and NUCB2/nesfatin–1 (NES)/NUCB2 change in serum and the reproductive tissues of female and male rats with fructose-induced metabolic syndrome, and whether the levels of each hormone is correlated with the hormones involved with fertility. Experiments were conducted on 5-week-old Sprague-Dawley male and female rats assigned to either a control group or a MetS group. Controls were fed standard rat food and water ad libitum, while the MetS group was fed standard food with 10% (v/v) fructose solution added to their drinking water for 12 weeks with a 12/12 h photoperiod circle. Then, all animals were sacrificed after a one night fast. Peptides levels in the serum and reproductive tissues of rats were studied using the ELISA method while the immunoreactivity of reproductive system peptide hormones were shown by immunohistochemical staining method. Furthermore, the other biochemical parameters were measured using Konelab-60 equipment and infertility hormones were measured with Immulite2000. Fasting serum insulin, glucose, triglyceride, alanine aminotransferase (ALT), gamma glutamyl transpeptidase (GGT), low-density lipoprotein cholesterol (LDL-C), and total cholesterol (TC) levels were statistically significantly higher, and the amount of high density lipoprotein cholesterol (HDL-C) was significantly lower, in the MetS groups. Serum and tissue supernatant NES levels were significantly higher in the rats with MetS than the control group. Ghrelin, OBS and NES were expressed in the cytoplasm, concentrated around the apical parts of the epithelial cells in the reproductive tissues of the rats. The amounts of ghrelin were lower in the reproductive tissues of the animals with MetS, while NES levels in the same tissues increased. Obestatin also decreased, though not in the seminal glands.
[Show abstract][Hide abstract] ABSTRACT: Objective:
Malignant mesothelioma (MM) is a primary malignant tumor developing from mesothelial cells lining the serosal surfaces and particularly the pleura, and has a very poor prognosis. It may display a variety of histological patterns and has a wide spectrum of cytomorphological characteristics, causing problems in its differential diagnosis from lung adenocarcinomas and sometimes from benign mesothelial proliferations. Immunohistochemical examination is the most useful method for this distinction. In our study, we aimed to determine the value of glucose transporter isoform-1 (GLUT-1) and K homology domain-containing protein (KOC) markers in the differential diagnosis of reactive mesothelial hyperplasia, malignant mesothelioma and lung adenocarcinoma.
Material and method:
Our study included 30 samples of malignant mesothelioma, 30 samples of pulmonary adenocarcinoma and 30 samples of reactive mesothelial hyperplasia selected from the archives of the Fırat University Hospital's Pathology Department Laboratory. The samples were applied GLUT-1 and KOC markers by immunohistochemistry and the place of these markers in the differential diagnosis was examined.
GLUT-1 was found positive in 80% of malignant mesothelioma cases, 83.3% of adenocarcinoma cases and 6.6% of reactive mesothelial hyperplasia cases. KOC was positive in 83.3% of malignant mesothelioma cases, 76.6% of adenocarcinoma cases and 46.6% of reactive mesothelial hyperplasia cases. There was no statistically significant difference between malignant mesothelioma and lung adenocarcinoma cases in terms of the diffuseness and intensity of staining with GLUT-1, whereas a significant difference was established when these groups were compared with reactive mesothelial hyperplasia cases. However, the KOC staining diffuseness and intensity results were similar to those obtained with GLUT-1.
In conclusion, GLUT-1 and KOC markers do not differentiate malignant mesotheliomas from pulmonary adenocarcinomas but can be useful in differentiating reactive mesothelial hyperplasia from malignant mesothelioma and lung adenocarcinoma.
No preview · Article · May 2013 · Turk Patoloji Dergisi
[Show abstract][Hide abstract] ABSTRACT: Inflammatory myofibroblastic tumor is an uncommon spindle cell tumor, occurring mainly in children and young adults. It is an extremely rare cardiac tumor especially patients under 1 year. Although it is benign, the tumor may be very aggressive locally. The diagnosis of this unusual pediatric cardiac tumor without pathologic specimens is difficult. We report a rare case of inflammatory myofibroblastic tumors of the right ventricle in a 7-month-old girl presenting with respiratory distress and cyanosis.
No preview · Article · Dec 2012 · Congenital Heart Disease
[Show abstract][Hide abstract] ABSTRACT: This study aimed to examine the effects of CPB on salusin-α, salusin-β and apelin-36 bioactive peptides in people who are planned to undergo coronary artery bypass graft (CABG) operation due to coronary artery disease and to explore whether these peptides are produced in human aortic, saphenous and arterial tissues. The study included age and BMI matched 15 patients who underwent CABG operation by CPB. In order to determine salusin-α, salusin-β and apelin-36 levels, venous blood samples were collected before induction of anesthesia (T1), before CPB (T2), 5min before the removal of cross-clamp (T3), 5min after the removal of cross-clamp (T4), upon arrival in the intensive care (T5), at postoperative 24th hour (T6) and 72nd hour (T7). Salusin and apelin expressions of the tissues were shown by immunohistochemical method. Peptide amounts of sera and tissues were measured using ELISA. Salusins production by vessels occurs in fibroblast cells of the media in the aorta and smooth muscle cells of the media in the LIMA and saphena. Apelin is produced by endothelial cells of the intima and fibroblast cells of the media in the aorta and by smooth muscle cells of the media in the LIMA and saphena. Changes in the levels of salusin-β and apelin-36 were significant during CPB. Salusin-α, salusin-β and apelin-36 are locally synthesized in the arteries and veins. Salusins and apelin-36 might be important markers in the CPB, and also that salusin-β was more specific in comparison to salusin-α.
[Show abstract][Hide abstract] ABSTRACT: T lymphocytes induce the transformation of fibroblasts into myofibroblasts, the main mediators of fibrogenesis. The inosine 5'-monophosphate dehydrogenase inhibitor mycophenolate mofetil (MMF) and the anti-CD25 monoclonal antibody daclizumab (DCZ) have been reported to suppress the proliferation of T lymphocytes.
To evaluate the preventive effects of MMF and DCZ in early stages of bleomycin (BLM)-induced scleroderma.
This study involved five groups of Balb/c mice (n = 10 per group). Mice in four of the groups were injected subcutaneously (SC) with BLM [100 μg/day in 100 μL phosphate-buffered saline (PBS)] for 4 weeks; the remaining (control) group received only 100 μL PBS. Three of the BLM-treated groups also received either intraperitoneal MMF 50 or 150 mg/kg/day, or SC DCZ 100 μg/week. At the end of the fourth week, all mice were killed, and blood and tissue samples were obtained for further analysis.
In the BLM-treated group, increases were seen in inflammatory-cell infiltration, α-smooth muscle actin-positive (α-SMA+) fibroblastic cell count, tissue hydroxyproline content, and dermal thickness. Dermal fibrosis was histopathologically prominent. In BLM-treated mice also given MMF or DCZ, inflammatory-cell infiltration, tissue hydroxyproline content and dermal thickness were decreased. In the MMF groups, decreases were also noted in α-SMA+ fibroblastic cell count.
In this BLM-induced dermal fibrosis model, MMF and DCZ treatments prevented the development of dermal fibrosis. Further studies are needed to evaluate whether targeting T lymphocytes is effective in resolving pre-existing fibrosis in human scleroderma.
No preview · Article · Jan 2012 · Clinical and Experimental Dermatology
[Show abstract][Hide abstract] ABSTRACT: Pleomorphic carcinoma of the lung is a subtype of sarcomatoid carcinoma and essentially classified as a poorly-differentiated, non-small cell lung carcinoma. Being a very rare tumor, it constitutes 0.3-1.3% of all malignancies of the lung. Cytology reveals malignant fusiform and/or giant cells, accompanied by malignant epithelial elements like squamous cell, adeno or large cell carcinoma. Our case, a 76-year-old female patient, presented with chest and back pain. Thoracic CT showed a well-demarcated solid mass of 5x3 cm located peripherally in the left upper lobe of the lung. Trans-thoracic fine needle aspiration cytology showed atypical cells with a biphasic character in a myxoid matrix. It was noted that of these, some were poorly-demarcated fusiform cells with oval nuclei and marked nucleoli, while others were epithelial cells with eccentrically placed nuclei, large cytoplasms and macronucleoli. The patient was diagnosed as pleomorphic carcinoma on the basis of these findings, and the cytological diagnosis was confirmed by histopathology. Pleomorphic carcinoma is a poorly-differentiated non-small cell lung carcinoma, which poses diagnostic difficulties. As it is rare, it lacks decisive diagnostic criteria and has cytological characteristics resembling those of other lung tumors.
No preview · Article · Jan 2012 · Turk Patoloji Dergisi
[Show abstract][Hide abstract] ABSTRACT: The aims of the present study were to examine ghrelin expression in serum and gastrointestinal tract (GIT) tissues, and to measure tissue ghrelin levels and obesity-related alterations in some serum biochemical variables in rats with diet-induced obesity (DIO). The study included 12 male rats, 60 days old. The rats were randomly allocated to two groups (n = 6). Rats in the DIO group were fed a cafeteria-style diet to induce obesity, while those in the control group were fed on standard rat pellets. After a 12 week diet program including an adaptation period all rats were decapitated, tissues were individually fixed, ghrelin expression was examined by immunohistochemistry , and tissue and serum ghrelin levels were measured by radioimmunoassay. Serum biochemical variables were measured using an autoanalyzer. When the baseline and week 12 body mass index and GIT ghrelin expression were compared between DIO and control rats, BMI had increased and ghrelin expression decreased due to obesity. The RIA results were consistent with these findings. Serum glucose, LDL cholesterol, and total cholesterol levels were elevated and HDL cholesterol significantly decreased in the DIO group. A comparison of GIT tissues between the control and obese groups demonstrated that ghrelin was decreased in all tissues of the latter. This decrease was brought about a decline in the circulating ghrelin pool. This suggests that rather than being associated with a change in a single tissue, obesity is a pathological condition in which ghrelin expression is changed in all tissues.
No preview · Article · May 2011 · Molecular and Cellular Biochemistry
[Show abstract][Hide abstract] ABSTRACT: Pulmonary hamartoma is the most common benign neoplasm of the lung, but the cystic form is very rare. This report presents the case of a 31-year-old woman with two cystic pulmonary lesions. She was radiologically and clinically diagnosed to have multiple ruptured hydatid cysts, and underwent a thoracotomy. The pathological investigation revealed that these lesions were cystic chondroid hamartomas, and one of the cysts was colonized by Aspergillus. Multilocular pulmonary cystic hamartomas are exceptionally rare and should be differentiated from other cystic pulmonary lesions. This is the first case of cystic pulmonary hamartomas colonized by Aspergillus species.
[Show abstract][Hide abstract] ABSTRACT: Endometrium carcinoma ranks fourth among female carcinomas. Therefore, early diagnosis of endometrium pre-malignant lesions is emphasised, and attempts are made to identify the risk factors. Since hyperplasias, particularly those with atypia, are held responsible for the development of the most common endometrium carcinomas, it is important to definitely distinguish between well-differentiated carcinomas and hyperplasia with atypia. In the present study, we aimed to explore whether ghrelin expression had a role in distinguishing between benign, pre-malignant and malignant lesions of endometrium.
Tissue ghrelin expressions of a total of 60 cases, who were diagnosed in the Pathology Department Laboratory of Firat University Medical School, and of whom 10 were in the proliferation phase, 10 had simple hyperplasia without atypia, 10 had simple hyperplasia with atypia, 10 had complex hyperplasia without atypia, 10 had complex hyperplasia with atypia and 10 had endometrioid carcinoma cases, were examined using immunohistochemical method. Additionally, tissue samples were homogenised to analyse tissue ghrelin levels in the supernatants according to RIA method. Samples from the parotid glands were used as positive control for ghrelin. Cells that exhibited cytoplasmic staining with ghrelin antibody were evaluated as positive.
Immunohistochemical examination showed that ghrelin expression increased markedly in the proliferation phase, relative to hyperplasias and carcinoma. These results were parallel to ghrelin levels in tissue supernatants. Immunohistochemical and RIA analysis results indicate that ghrelin expression either markedly decreases or is entirely depleted in endometrial carcinomas.
Therefore, we think that ghrelin expression can be useful in differentiating not only endometrium carcinomas from benign lesions but also complex hyperplasias with atypia, which pose diagnostic difficulties.
No preview · Article · Mar 2011 · Gynecological Endocrinology
[Show abstract][Hide abstract] ABSTRACT: Cytological examination of pleural fluids is a fast, efficient and non-invasive diagnostic method. Identification of malignant effusions bears critical importance in treatment and prognosis. The aim of this study was to investigate the distribution of cytopathologic diagnoses in pleural effusions and the cyto-histopathologic correlation rates.
A total of 298 pleural fluid cases diagnosed from 2008 to 2009 in our laboratory were retrospectively identified as the study group. Evaluation results were classified in 5 groups as inconclusive, benign, atypical, suspicious and malignant, and compared with the biopsy results.
Of the total 298 pleural fluid cases, 114 (38.3%) were females and 184 (61.7%) were males. The age range was between 15 and 89 with a mean value of 58.4 +/- 17.8. Of the cases, 3 (1%) were diagnosed as inadequate, 246 (82.6%) benign, 8 atypical, 10 suspicious, and 31 malignant by cytology. Among the cases who were diagnosed as malignant, 24 (8.1%) were reported as metastatic carcinoma and 7 (2.3%) as malignant mesothelioma. Of the 8 cases reported as atypical, biopsy results of 2 showed malignant mesothelioma, and of the 10 cases reported as suspicious, 1 case whose biopsy result was obtained was diagnosed as epidermoid carcinoma metastasis.
Cytological examination is the most valuable diagnostic method for pleural effusions which may have various etiological causes. The most common cause of pleural effusions in our region is metastatic carcinomas including those from the lung, breast and ovarian tumors, followed by malignant mesothelioma.
No preview · Article · Jan 2011 · Turk Patoloji Dergisi
[Show abstract][Hide abstract] ABSTRACT: Ghrelin, an endogenous ligand of the growth hormone secretagogue receptor, has been identified in mammals, fish, amphibians, birds, reptiles and some plants. The present investigation was designed to determine whether ghrelin is present in the appetite-stimulating plants Syzygium aromaticum and Salvadora persica, using IHC (immunohistochemistry) to indicate the location of the peptide and ELISA to measure the concentration. ELISA demonstrated that a ghrelin-like substance was present at concentrations of 4070.75±664.67 and 75.25±24.49 pg/mg in the tissues of flower bud of S. aromaticum and branch of S. persica, respectively. The concentration of ghrelin in human salivary gland tissue was 436.00±95.83 pg/mg. Ghrelin was predominantly localized to the T (trachea) and PCs (parenchyma cells) in the flower bud of S. aromaticum. However, no ghrelin immunoreactivity was observed in the PC or T of the branch of S. persica. The evolutionary role of this peptide hormone in plants and animals suggests that they have evolved in a more similar way than previously thought.
Full-text · Article · Oct 2010 · Cell Biology International
[Show abstract][Hide abstract] ABSTRACT: Primary pleural liposarcoma is extremely rare, and its most common subtype is the myxoid subtype. To the best of our knowledge, the number of cases reported up to now is <20. It is primarily a disease ofindividuals 50 years of age and it is more common in men. It has no specific symptoms except for cough and chest pain.
A 56-year-old female presented complaining of chest pain and dyspnea for 4 months. An increase in homogeneous density on the left hemithorax, from the apex to basal area, with blurring of the borders of the heart and the diaphragm, was revealed by lung radiography. Thoracic computed tomography revealed a diffuse pleural effusion causing compressive atelectasis at the base of the left hemithorax. Cytologic examination of the pleural fluid demonstrated tumoral cells forming papillary structures with complex branching and fine fibrovascular core. The tumor cells were relatively uniform and had vacuolar cytoplasm with hyperchromatic nuclei. Upon finding CA-125 positive in the immunocytochemical examination, the case was reported as "consistent with malignancy, possibility of ovarian origin, cannot be excluded". As the gynecologic examination was normal, exploratory thoracotomy was performed to take a biopsy. The case was diagnosed as liposarcoma (myxoid type) by histopathologic examination.
Cytologic diagnosis of pleural liposarcoma is difficult due to its rarity and resemblance to malignant mesothelioma. Cytologic properties of liposarcoma in pleural fluid should be well known and considered in the differential diagnosis.
[Show abstract][Hide abstract] ABSTRACT: The underlying molecular mechanism of carcinogenesis in oral squamous cell carcinoma (OSCC) is poorly understood and appears to be controlled on many genetic, environmental, and hormonal factors. Obestatin and ghrelin, two recently discovered hormones, are co-expressed in endocrine cells. The purpose of this investigation was to examine the immunohistochemical features of OSCCs in relation to the tissue concentration of ghrelin and obestatin. The association between OSCC and Epstein Barr Virus (EBV) status was also explored. The expression of ghrelin and obestatin was examined by immunohistochemistry and immunoassay in oral biopsy specimens: 10 benign squamous epithelial cell samples, 10 microinvasive squamous cell carcinomas, and seven well-differentiated and seven poorly differentiated OSCCs. The presence of EBV was evaluated in these samples using immunohistochemistry. The concentrations of ghrelin and obestatin in tissue homogenates were measured by RIA and ELISA, respectively. Squamous cell carcinomas and benign tissue samples were positive for anti-EBV antibody, and obestatin and ghrelin were shown to be co-expressed in all stratified squamous epithelium samples. Expression of ghrelin and obestatin was decreased or absent in OSCCs in relation to the invasiveness of the carcinoma; ghrelin and obestatin levels in cancerous tissue homogenates were lower than in benign tissue homogenates. These results indicate that the concentrations and distribution of immunoreactive obestatin and ghrelin might be helpful in distinguishing OSCC from benign tumors. Maintaining normal levels of these hormones might be required for regulation of normal cell division. However, detailed studies will be required for better understanding of the complex mechanism of carcinogenesis relating to OSCCs.
No preview · Article · Jun 2010 · Molecular and Cellular Biochemistry
[Show abstract][Hide abstract] ABSTRACT: This study was designed to measure the levels of chromogranin A (CgA), ghrelin and obestatin in serum and saliva (including CgA expression in healthy tissue) in epileptic patients to determine any significant differences between these patients and healthy controls. Samples were obtained from a total of 91 subjects: 10 newly-diagnosed primary generalized epilepsy (PGE) patients who had started treatment with valproic acid and phenytoin for seizure control; 18 PGE patients who were previously and currently receiving treatment with valproic acid and phenytoin for seizure control; 37 patients with partial epilepsy (PE) (simple, n=17 or complex, n=20) who had been and were still being treated with carbazebime for seizures; and 26 healthy controls. CgA immunoreactivity in healthy salivary gland was analyzed by immunohistochemistry and ELISA. The levels of CgA, total ghrelin and obestatin in serum and saliva were measured by ELISA. The results revealed that normal salivary gland produces its own CgA. Before treatment, CgA levels in saliva and serum were significantly greater in patients newly-diagnosed with PGE than controls. Ghrelin and CgA concentrations were also greater in PGE patients previously or currently treated with drugs, and in patients with simple or complex partial epilepsy (PE) previously or currently treated with drugs, than in healthy normal controls. In conclusion, salivary concentrations of CgA, ghrelin and obestatin were similar to their serum levels, so saliva might be a desirable alternative to serum for measuring these hormones because it is easy and painless to collect.