Is development of high-grade gliomas sulfur-dependent?

We characterized γ-cystathionase, rhodanese and 3-mercaptopyruvate sulfurtransferase activities in various regions of human brain (the cortex, thalamus, hypothalamus, hippocampus, cerebellum and subcortical nuclei) and human gliomas with II to IV grade of malignancy (according to the WHO classification). The human brain regions, as compared to human liver, showed low γ-cystathionase activity. The activity of rhodanese was also much lower and it did not vary significantly between the investigated brain regions. The activity of 3-mercaptopyruvate sulfurtransferase was the highest in the thalamus, hypothalamus and subcortical nuclei and essentially the same level of sulfane sulfur was found in all the investigated brain regions. The investigations demonstrated that the level of sulfane sulfur in gliomas with the highest grades was high in comparison to various human brain regions, and was correlated with a decreased activity of γ-cystathionase, 3-mercaptopyruvate sulfurtransferase and rhodanese. This can suggest sulfane sulfur accumulation and points to its importance for malignant cell proliferation and tumor growth. In gliomas with the highest grades of malignancy, despite decreased levels of total free cysteine and total free glutathione, a high ratio of GSH/GSSG was maintained, which is important for the process of malignant cells proliferation. A high level of sulfane sulfur and high GSH/GSSG ratio could result in the elevated hydrogen sulfide levels. Because of the disappearance of γ-cystathionase activity in high-grade gliomas, it seems to be possible that 3-mercaptopyruvate sulfurtransferase could participate in hydrogen sulfide production. The results confirm sulfur dependence of malignant brain tumors.

Transthyretin amyloidosis with pulmonary involvement in a patient with monoclonal gammapathy.

Pulmonary involvement in the course of systemic senile amyloidosis caused by non-mutated transthyretin is rarely described. We report on concomitant monoclonal gammapathy of undermined significance (MGUS) and amyloidosis with non-mutated transthyretin with diffuse lesions in lung parenchyma. A female patient, 67 years old, was admitted with dyspnoea, malaise, weight loss, and disseminated radiological lesions in the lungs. On lung HRCT, signs of pulmonary hypertension, alveolar and interstitial involvement, with thickening of septal lines were found. Echocardiography revealed severe pulmonary hypertension, and electromyography revealed sensoromotoric polyneuropathy with axon and myelin damage. Pathological assessment of lung specimens revealed nodular deposits of amyloid in the bronchial walls and lung parenchyma Congo red staining was positive. Specimens of colon mucosa confirmed amyloidosis. Stainings for AA, AL and beta2-microglobulin were negative but were positive for transthyretin. Bone marrow trepanobiopsy indicated monoclonal gammapathy of MGUS type; Congo red staining was positive. Transthyretin amyloidosis with vascular involvement, particularly of arteriovenous anastomoses, including pulmonary vessels and an insignificant amount of AL protein (perhaps secondary imbibition with AL protein from serum) was diagnosed in amyloid deposits. No mutations of the transthyretin gene (exon 1,2,3,4) were found. The patient was treated with methylprednisolone, melphalan and then with cyclophosphamide. Radiological examinations performed 1 and 2 month/s after initiation of therapy showed progression of pulmonary lesions. The patient died one month later; an autopsy was not performed.

Pulmonary arteriovenous malformations in children and young adults.

Authors observed five cases of lung arteriovenous malformations in children and young adults. Clinical data and morphological pictures of these lesions were presented.

Cribriform adenocarcinoma of the tongue.

We report a case of a 73-year-old female with a tumour of the tongue, operated with two relapses. A single metastasis to the lymph node was present. Currently, the patient is alive without evidence of disease. The histological diagnosis of cribriform adenocarcinoma of the tongue was rendered. The differential diagnosis of adenocarcinomas of the tongue is discussed.

Pulmonary acinic cell carcinoma (Fechner tumour) with carcinoid component. A case report.

The authors present a very rare case of primary lung acinic cell carcinoma with carcinoid component in a 53-year-old man.

Immunohistochemical investigation of selected endothelial markers in pulmonary epithelioid haemangioendothelioma.

Epithelioid haemangioendothelioma (EH) is a very rare neoplasm. It is assumed that these tumours derive from vascular endothelial cells. The aim of the study was to identify the immunohistochemical profile of tumour cells in lung EH. The unique material consisting of eight cases of lung EH was collected and examined by immunohistochemistry with three antibodies: CD-31, CD105, both marking vascular endothelial cells, and D2-40, marking lymphatic endothelial cells. In all cases, the cytoplasm of tumour cells showed widespread expression of CD-31. Reaction with CD-105 antibody gave a positive result mainly in the cytoplasm of tumour cells located at the periphery of tumours, especially in highly cellular neoplasms containing spindle cells. Reaction with D2-40 antibody in most cases was negative. The presence of a few capillary vessels with positive endothelial cells was revealed in two cases at the periphery of the tumour. Only in one case of a highly cellular tumour did a small portion of spindle cells show a positive reaction to D2-40. The above studies confirmed the origin of EH mainly from vascular endothelial cells. Medications inhibiting vascular endothelial growth factors can be considered as a treatment option for multifocal EH that does not qualify for surgical intervention.

Exogenous lipoid pneumonia (oil granulomas of the lung).

The authors observed three cases of exogenous lipid pneumonia clinically suspected of lung carcinoma. Histological examination of material after thoracotomy gave the possibility of correct diagnosis. The lesions in lungs were characteristic granulomas around lipid material and with surrounding advanced fibrosis.

TGF-beta1 in bronchoalveolar lavage fluid in diffuse parenchymal lung diseases and high-resolution computed tomography score.

INTRODUCTION: In the pathogenesis of diffuse parenchymal lung diseases (DPLDs), growth factors, including transforming growth factor beta1 (TGF-beta1), are responsible for cell proliferation, apoptosis, chemotaxis, and angiogenesis, and also for the production and secretion of some components of the extracellular matrix. OBJECTIVES: The aim of the study was to evaluate correlations in DPLDs between TGF-beta1 levels in bronchoalveolar lavage (BAL) fluid and high-resolution computed tomography (HRCT) score. PATIENTS AND METHODS: The study was performed in 31 DPLD patients in whom a selection of lung segments with high and low intensity of abnormalities was estimated by HRCT score. All patients underwent BAL with TGF-beta1 measured by an enzyme immunoassay in BAL fluid and video-assisted thoracic surgery lung biopsy from both selected segments. RESULTS: All 31 patients were diagnosed, and based on histopathology, they were classified into 2 groups: idiopathic interstitial pneumonia (usual interstitial pneumonia - 12, nonspecific inter stitial pneumonia - 2, cryptogenic organizing pneumonia - 2, and desquamative interstitial pneumonia - 1) and granulomatous disease (sarcoidosis - 7, extrinsic allergic alveolitis - 5, and histiocytosis X - 2). The final analysis was performed in 28 patients who showed nonhomogenous distribution on HRCT. TGF-beta1 levels in BAL fluid were significantly higher in the areas with high intensity of abnormalities assessed by HRCT score (P = 0.018, analysis of variance). These levels were not different between the groups, but a trend towards higher levels in idiopathic inter stitial pneumonia was observed. CONCLUSIONS: The results confirm that TGF-beta1 may be a good but not specific marker of fibrosis in DPLDs. A significant positive correlation between TGF-beta1 levels in BAL fluid and the HRCT score was observed.

Papillary adenoma of the lung.

The report presents a very rare case of papillary adenoma of the lung in a 61-year old man, described for the first time in the Polish literature.

Endothelial neoplasms of the lungs.

In the period of 20 years, the author had an opportunity to observe 14 cases of endothelial neoplasms of the lungs. The group consisted of eight women and six men, differing in age from 18 to 75 years. These cases presented as two subsets: low grade malignancy epithelioid hemangioendothelioma (PEH) (12 cases--eight women and four men, aged between 18 and 75 years) and highly malignant endotheliosarcoma seen in two men, 28 and 47 years old. The patients with epithelioid hemangioendothelioma reported to hospitals mainly because routine chest X-ray examinations accidentally revealed nodular lesions involving both lungs. Histological examination of the material collected from the bronchi and sputum was negative for neoplasm. The patients were suspected of suffering from tuberculosis or sarcoidosis and in some cases therapy was even initiated. Correct diagnoses were established based on histological examinations of material from the lesions taken during thoracotomy or thoracoscopy. In these cases, the course of the disease was slow and the patients were in a good condition for many years. In one case (a 27-year old woman), the diagnosis was possible after autopsy. The morphological appearance of these tumors is very characteristic. In case of any doubts, we can perform immunohistochemical examinations using endothelial markers, mainly CD34, possibly CD31 or factor VIII. Highly malignant endothelial sarcomas were seen in two men with a poor clinical status; one of them died shortly after histological diagnosis had been established based on material taken during a thoracotomy. In the second case, the diagnosis was possible on autopsy The prognosis for patients with these highly malignant tumors is highly unfavorable.

Histiocytic necrotizing lymphadenitis without granulocytic infiltration (the so called Kikuchi-Fujimoto disease).

The report describes five cases of a rare disorder--necrotizing lymphadenitis--diagnosed in Polish patients in the Department of Pathomorphology, Collegium Medicum, Jagiellonian University, Krakow, in the years 1993-2006. The disease was firstly described by Kikuchi and Fujimoto in the Oriental population of Japan in 1972 and for this reason it is called Kikuchi-Fujimoto disease (or Kikuchi lymphadenitis). Its characteristic histological picture includes necrosis without granulocytic infiltrate surrounded by plasmocytoid monocytes, histiocytes (CD68+, lysozyme+, myeloperoxidase+) and immunoblasts, sometimes with atypia, with concomitant lymphocytes, predominantly cytotoxic T CD8+. The histology together with the rare occurrence of the disease in Poland may be a considerable diagnostic challenge for a pathologist, leading to misdiagnosing the lesion as a neoplastic process (malignant lymphoma).

Gastric solid glomus tumor and multiple glomangiomyomas of the large bowel with intravascular spread, multifocal perivascular proliferations and liver involvement.

The authors present a case of multiple glomus tumors (GTs) of the gastrointestinal tract, representing the type of a gastric glomus tumor proper and large bowel glomangiomyomas with myopericytoma-like features, observed in a 46-year old female, with multifocal perivascular proliferations of primitive cells and hepatic involvement. Histologically, the multilobular gastric tumor and hepatic lesions corresponded to a typical glomus tumor, while the tumor situated in the transverse colon, up to 7 cm in diameter, presented as a glomangiomyoma infiltrative (with myopericytoma-like foci), and satellite tumors in the large bowel mucosa, 0.5-0.7 cm in diameter, represented small glomangiomyomas. In addition, the patient demonstrated two types of concomitant vascular lesions: 1/ intravascular spread in the form of neoplastic plugs that obliterated the lumen of medium-size veins, and 2/ microscopic perivascular proliferation of primitive, small cells seen in the vicinity of the main tumor and in the adjacent adipose tissue. The patient has been ill for 2.5 years; she has been subjected to a partial colectomy with a resection of the small intestinal loop, greater omentum and the right ovary, followed by chemotherapy. At present, she is stable, and the infiltration--especially in the epigastric region--has decreased. The picture may confirm the theory that multiple GTs develop in association with multifocal proliferation of perivascular stem cells, as well as that their ability to penetrate into the lumen of large vessels gives origin to satellite tumors, which are not necessarily metastatic. It seems that at present, the group of perivascular SMA+ tumors may include infantile-type myofibromatosis in adults, myopericytoma, glomangio(myo)pericytoma, glomangiomyoma, glomus tumor proper, and glomangioma. Most likely, also some tumors previously classified as hemangiopericytomas belong to this group. The distinctive feature present in at least some of the above listed perivascular tumors is their synchronous or metachronous growth in a particular region and their ability to occupy intravascular space as nodules or solid bands, which in turn may give origin to satellite tumors. Multifocal lesions associated with a short survival in a given patient will obviously support the presence of metastatic disease. In the remaining cases, determination whether the patient has metastatic disease requires deep consideration and caution, also while deciding on treatment to be employed.

The transcervical extended mediastinal lymphadenectomy versus cervical mediastinoscopy in non-small cell lung cancer staging.

OBJECTIVE: To compare the diagnostic yield of the transcervical extended mediastinal lymphadenectomy (TEMLA) and the cervical mediastinoscopy (CM) in detecting metastatic mediastinal lymph nodes in NSCLC patients. METHODS: Prospective, randomized, single-blind clinical study. RESULTS: There were 41 NSCLC patients enrolled in the study; 21 were randomized to the TEMLA group and 20 to the cervical mediastinoscopy group. The TEMLA revealed mediastinal metastases in 7 patients, and mediastinoscopy in 3. In the TEMLA group one patient out of the 14 with negative nodes was finally found unfit for surgery, and in the remaining 13 lung resections with mediastinal dissection were performed. In the mediastinoscopy group one patient out of the 17 with negative nodes was finally found unfit for surgery and another one refused surgery, so in 15 of them lung resections with mediastinal dissection were performed. In no patient in the TEMLA group did the pathological examination of the operative specimen reveal metastatic lymph nodes, whereas in the mediastinoscopy group metastatic nodes were found in 5 patients. The number of false negative results was significantly greater in the mediastinoscopy group (5 vs 0, p=0.019), and the difference was the reason for terminating the randomization before reaching the initially planned number of 100 patients. The sensitivity of mediastinoscopy was 37.5% and its negative predictive value was 66.7%, compared to 100% and 100% in the TEMLA group. The comparison of the time of the operation, blood loss, complications, postoperative pain and the use of analgetics has shown significant differences between groups only regarding the operative time and the pain intensity, being greater in the TEMLA group. CONCLUSIONS: 1. The sensitivity and the NPV of the TEMLA in detecting mediastinal metastases in NSCLC are significantly greater than those of cervical mediastinoscopy. 2. The invasiveness of TEMLA and mediastinoscopy does not significantly differ, except for the postoperative pain.

Effect of bilateral mediastinal lymphadenectomy on short-term pulmonary function.

OBJECTIVE: To assess if the bilateral mediastinal lymphadenectomy results in lymphatic congestion in the lungs producing clinically significant impairment of respiratory function. METHODS: In the prospective, randomized, double-blind clinical study, non-small cell lung carcinoma patients underwent preoperatively mediastinoscopy or the transcervical extended mediastinal lymphadenectomy (TEMLA). In both groups, the blood gas analysis and spirometry were measured preoperatively and on the 1st, 3rd, and 5th postoperative day, and the carbon monoxide diffusing capacity of the lung (DLCO) and lung compliance were measured preoperatively and on the 3-5 postoperative day. Any respiratory complications were also recorded. RESULTS: Forty-one patients were randomized: 21 to the TEMLA group and 20 to the mediastinoscopy group. There was no significant difference of the baseline and the 1st, 3rd, and 5th day measurements of vital capacity and forced expiratory volume (FEV1) (p>0.98), pH, pO(2), pCO(2), standard bicarbonates and base excess (p>0.31), nor significant difference of baseline and 3-5 day measurements for DLCO (p=0.91) and lung compliance (p=0.38). The incidence of respiratory insufficiency was not significantly different (p=0.51). CONCLUSIONS: (1) Complete excision of mediastinal lymph nodes stations 1, 2R, 2L, 3A, 4R, 4L, 5, 6, 7, and 8 (TEMLA) is not associated with greater incidence of respiratory insufficiency comparing with standard mediastinoscopy. (2) The TEMLA procedure does not produce greater alterations in spirometry, blood gas analysis, DLCO and lung compliance comparing with standard mediastinoscopy.

Tissue microarray FISH applied to colorectal carcinomas with various microsatellite status.

Colorectal carcinoma is etiopathologically heterogenic. It may develop through a sequence of mutations leading to chromosome instability or be a result of defects in DNA repair mechanisms manifested by microsatellite instability of varying degrees. Colorectal carcinoma can thus be classified into microsatellite-stable (MSS), highly microsatellite unstable (MSI-H) and intermediate low-level microsatellite unstable (MSI-L) groups. Fluorescent hybridization in situ (FISH) is a method of detecting specific sequences of nucleic acids that is based on specific bonding of a fluorescent marker-associated probe and specific DNA fragment. The material consisted of 146 non-selected cases of colorectal carcinoma patients operated on at First Chair of General Surgery, Collegium Medicum, Jagiellonian University in Cracow, Poland. Following a standard histopathological evaluation, tissue microarrays were prepared using a Tissue MicroArray Builder, and FISH was performed employing probes specific for chromosomes 1, 8, 17 and 18. Microsatellite instability was evaluated in frozen material using the PCR reaction with gel and capillary electrophoresis. The mean number of signals obtained for chromosome 1 in the entire material was 2.06, while the corresponding mean values in the MSS group equaled 2.07, in the MSI-L group - 2.07, and in the MSI-H group - 2.01. The mean number of signals for chromosome 17 in the entire material was 2.1, in the MSS group - 2.11, in the MSI-L group - 2.13, and in the MSI-H group - 2.01. The number of signals for chromosome 18 in the entire material was 2, in the MSS group - 2, in the MSI-L group - 2, and in the MSI-H group - 2. The means number of signals for chromosome 8 in the entire material was 2.07, in the MSS group - 2.08, in the MSI-L group - 2.01, and in the MSI-H group - 2. These differences are not sufficient for distinguishing colorectal carcinoma molecular forms.

Correlation of microsatellite status, proliferation, apoptotic and selected immunohistochemical markers in colorectal carcinoma studied with tissue microarray.

Colorectal carcinoma is a frequent malignant tumor, characterized by varying clinical course and response to treatment. At the molecular level, colorectal carcinomas are divided into tumors with chromosomal instability (microsatellite-stable, MSS), microsatellite instability (MSI-H) and low microsatellite instability (MSI-L). The method of tissue microarrays allows for combining materials originating from multiple patients into a single slide, what makes possible to simultaneously investigate large material for the presence of numerous, diversified markers. The study material consisted of 208 cases of colorectal carcinoma. Microsatellite instability was evaluated in frozen material employing the PCR reaction with gel and capillary electrophoresis. Following a standard histopathological assessment, tissue microarrays were prepared using a MTA-1 microarrayer (Beecher) and standard immunohistochemical reactions were performed to detect the presence of bcl-2, CDX-2, Ki67, MLH1, MSH2, MSH6, p16, p53 and survivin. Apoptotic cells were detected using the TUNEL method. The correlations between the reactions were investigated and differences in the expression of the investigated proteins noted in carcinomas with various degrees of microsatellite instability. The agglomeration analysis showed differences in patterns of expression between MSS, MSI-L and MSI-H carcinomas. The discriminant function analysis demonstrated that the MSI-H carcinomas were best differentiated by MLH1, survivin and Ki67 expression, while the MSI-L tumors differed from the remaining colorectal carcinomas by their apoptotic index, local tumor stage (pT), the presence of angioinvasion and mucin production.

Middle mediastinal thymoma of unusual pathologic type.

Among the many sites of ectopic thymoma development, the middle mediastinum is the rarest in the English language literature we found only two case reports. We present a case of 69-year-old woman with slow-growing, ectopic middle mediastinum thymoma of an very unusual histological type, neither classified in the Müller-Hermelink nor the World Health Organization (WHO) classification.

The relationship between MSI status and vessel density in colorectal carcinoma.

The development of new blood vessels is a prerequisite for progression of malignant neoplasms. Factors that induce neoangiogenesis include VEGF, VEGF-C, VEGF-D, PD-ECG, ANG-2, TSP-1, HIF-1 and HIF-2. From the etiopathogenetic viewpoint, colorectal carcinoma is heterogenic. It may develop via a sequence of mutations leading to chromosome instability or else result from DNA repair defects, which are manifested as microsatellite instability. The objective of the present investigations was the comparison of neoangiogenesis in microsatellite-stable colorectal carcinomas, as well as in tumors with low and high instability levels. The material included 71 surgical cases of colorectal carcinoma. Vessel density was assessed by immunohistochemical reactions to CD34 and vWf, calculating the number of vessel sections within the invasion margin, in visual fields selected at random, and within hot spots. Microsatellite instability was evaluated in frozen materials employing the PCR reaction with gel and capillary electrophoresis. In all the cases, the authors detected CD34+ and less numerous vWf+ vessels within the tumor and in its vicinity. In 45 cases, no microsatellite instability was found, in 13 cases, low level instability (MSI-L) was observed, and in another 13 - high microsatellite instability (MSI-H). Some differences in vessel density were noted between the above groups, yet they were not statistically significant. On the other hand, the authors observed more numerous CD34+ vessels in cases with metastases to the regional lymph nodes. In conclusion, it is suggested that neoangiogenesis in sporadic colorectal carcinoma is directly related to metastatic potential, but not to MSI status.

High thymidylate synthase expression is typical for sporadic MSI-H colorectal carcinoma.

Colorectal carcinoma is etiopathologically heterogenic. It may develop through a sequence of mutations leading to chromosome instability or be a result of defects in DNA repair mechanisms manifested by microsatellite instability. Carcinomas of this type are supposed to be characterized by a better prognosis and a different response to chemotherapy. The main target of 5-fluorouracil (5-FU) treatment is thymidylate synthase (TS). High TS expression has been identified as promoting resistance to 5-FU. The objective of the present investigation was to determine whether microsatellite instability is associated with thymidylate synthase expression. Ninety-eight cases of colorectal carcinoma were studied. Microsatellite instability was evaluated in frozen material employing the PCR reaction with gel and capillary electrophoresis. TS expression levels were assessed in preparations stained immunohistochemically using a semiquantitative method on a scale with scores from 0 to 3. The MSI-H phenotype was detected in ten cases, MSI-L in 16, and MSS in 72. The mean TS expression score was 1.79. In the MSS group, the mean TS expression score was 1.69, in the MSI-L group the mean TS expression score was 1.73, and in the MSI-H group the mean TS expression score was 2.67. The differences between MSI-H and MSS/MSI-L were statistically significant (p<0.0002 and p<0.004, respectively). The results may explain the different response of MSI-H carcinomas to 5-FU treatment.

Choriocarcinoma within the third trimester placenta--a case report.

A small focus of choriocarcinoma has been incidentally found within an otherwise normal, third trimester placenta which was sent to the pathomorphological examination because of the stillbirth of the child at 31st week of gestation. The autopsy of the child was not performed. Macroscopically focus of choriocarcinoma has looked as a small infarct and was indistinguishable from the second lesion, which was microscopically confirmed to be a real infarct. beta-hCG level was increased at the time of delivery, but come back to normal very quickly. There were no metastases in the mother at the time of delivery and the follow-up did not show any evidence of metastatic disease either.