Small extracellular vesicles containing arginase-1 suppress T-cell responses and promote tumor growth in ovarian carcinoma.

Tumor-driven immune suppression is a major barrier to successful immunotherapy in ovarian carcinomas (OvCa). Among various mechanisms responsible for immune suppression, arginase-1 (ARG1)-carrying small extracellular vesicles (EVs) emerge as important contributors to tumor growth and tumor escape from the host immune system. Here, we report that small EVs found in the ascites and plasma of OvCa patients contain ARG1. EVs suppress proliferation of CD4+ and CD8+ T-cells in vitro and in vivo in OvCa mouse models. In mice, ARG1-containing EVs are transported to draining lymph nodes, taken up by dendritic cells and inhibit antigen-specific T-cell proliferation. Increased expression of ARG1 in mouse OvCa cells is associated with accelerated tumor progression that can be blocked by an arginase inhibitor. Altogether, our studies show that tumor cells use EVs as vehicles to carry over long distances and deliver to immune cells a metabolic checkpoint molecule - ARG1, mitigating anti-tumor immune responses.

Role of WNT/ß-Catenin Pathway as Potential Prognostic and Predictive Factors in Renal Cell Cancer Patients Treated With Everolimus in the Second and Subsequent Lines.

BACKGROUND: The aim of the present study was to search for predictive and prognostic factors in patients with metastatic renal cell carcinoma (mRCC) treated with everolimus among the components of the WNT/ß-catenin pathway. PATIENTS AND METHODS: In a prospective, single-arm, phase II study, patients with mRCC received everolimus (10 mg/d) in a 30-day cycle. We performed a prospectively planned evaluation of the potential biomarkers of the WNT/ß-catenin pathway. RESULTS: The serum level of soluble E-cadherin (sE-cadherin) in patients with RCC was significantly greater than that in the controls (71.62 ± 22.28 pg/mL vs. 54.26 ± 10.317 pg/mL; P = .0069). After 2 cycles of everolimus therapy, we observed a significance increase in sE-cadherin (from 71.81 ± 21.18 pg/mL to 77.50 ± 28.212 pg/mL; P = .0151). The Dickkopf-1 protein levels in the study and control groups were not significantly different (P = .2135). The favorable independent predictors for everolimus therapy were normal lactate dehydrogenase level before treatment (hazard ratio [HR], 0.52; 95% confidence interval [CI], 0.28-0.98; P = .0443) and low sE-cadherin level (HR, 0.54; 95% CI, 0.29-0.98; P = .0422). On multivariate analysis, we observed that worse overall survival was seen in patients with a lower regression coefficient of sE-cadherin after 2 cycles of treatment (HR, 2.60; 95% CI, 1.23-5.52; P = .0128), an increased corrected calcium level (HR, 3.09; 95% CI, 1.21-7.88; P = .0180), and an increased lactate dehydrogenase level before treatment (HR, 1.98; 95% CI, 1.02-3.83; P = .0426). CONCLUSION: WNT/ß-catenin component expression in patients with mRCC had no effect on progression-free survival or overall survival. However, we found that the sE-cadherin level might interact with response to everolimus therapy, although confirmation in future studies is needed.

Computer approach to recognition of Fuhrman grade of cells in clear-cell renal cell carcinoma.

OBJECTIVE: To present a computerized system for recognition of Fuhrman grade of cells in clear-cell renal cell carcinoma on the basis of microscopic images of the neoplasm cells in application of hematoxylin and eosin staining. STUDY DESIGN: The applied methods use combined gradient and mathematical morphology to obtain nuclei and classifiers in the form of support vector machine to estimate their Fuhrman grade. The starting point is a microscopic kidney image, which is subject to the advanced methods of preprocessing, leading finally to estimation of Fuhrman grade of cells and the whole analyzed image. RESULTS: The results of the numerical experiments have shown that the proposed nuclei descriptors based on different principles of generation are well connected with the Fuhrman grade. These descriptors have been used as the diagnostic features forming the inputs to the classifier, which performs the final recognition of the cells. The average discrepancy rate between the score of our system and the human expert results, estimated on the basis of over 3,000 nuclei, is below 10%. CONCLUSION: The obtained results have shown that the system is able to recognize 4 Fuhrman grades of the cells with high statistical accuracy and agreement with different expert scores. This result gives a good perspective to apply the system for supporting and accelerating the research of kidney cancer.

Nucleolus detection in the Fuhrman grading system for application in CCRC.

The paper presents a method for nucleolus detection in images of nuclei in clear-cell renal carcinoma (CCRC). The method is based on the similarity of the nuclei image and the two-dimensional paraboloidal window function. The results of numerical experiments performed on almost 2600 images of CCRC nuclei have confirmed the good accuracy of the method. The developed algorithm will be used to accelerate further research in computer-assisted diagnosis of CCRC.

Study on breast carcinoma Her2/neu and hormonal receptors status assessed by automated images analysis systems: ACIS III (Dako) and ScanScope (Aperio).

UNLABELLED: Her-2/neu is overexpressed in 20-30% of breast cancer patients and is associated with a more aggressive disease. Identification of Her-2/c-erbB-2-neu overexpression is based on immunohistochemical [ihc] detection of protein and/or gene amplification in fluorescence in situ hybridization test (FISH). Also Estrogen receptors [ER] and Progesterone receptors [PR] are the prognostic and predictive biomarkers, recently analysed by ihc methods. Subjective, manual scoring of the ihc Her-2/neu expression and expression of the ER/PR reported as the percentage of immunopositive cells are the most common mode of interpretation among pathologists. Automated microscopy and computerised processing have provided increased accuracy in quantification and standardisation. THE AIMS OF OUR STUDY WERE: to evaluate the scoring reproducibility of Her-2 /neu ihc expression tested by two automated systems: ACIS (Dako) and ScanScope (Aperio); to estimate the ER/PR expression in ihc staining methods with different anti-ER/anti-PR antibodies (the monoclonal and the ER/PR pharmDx TM Kit) by the ACIS system. Her-2/neu ihc expression was measured in 114 primary invasive breast carcinomas by the manual and the automated scoring (ACIS and Aperio system). 106 slides stained ihc with two types of anti-ER/anti-PR antibodies entered the quantisation. The results of our investigations showed very high reproducibility of Her-2/neu scores in intra- and interobserver analysis by ACIS evaluation. The major concordance was present in strong 3+ ihc cases; very small discordance was shown by cases with low expression of Her-2/neu. The accuracy of scoring by the Aperio was little lower in comparison to ACIS but it might result from the smaller and variable series of samples analysed by Aperio. The concordance in scoring of two automated systems was 86.5% (p<0.0001; gamma=0.887); the discordance was referred only to the lower expression of Her-2/neu. The concordance in manual scoring performed by the single observer and the panel was 84.2% (p<0.0001, gamma = 0.99); the discordance comprised a few cases with strong expression (2+ vs 3+). Very high intra- and interobserver reproducibility of the ER/PR ihc measurements was present in the readers results (referred to the percentage of immunoreactive carcinomatous cell population in the breast carcinomas acc. to the ACIS algorithm). No differences were disclosed in the percentage of ER-immunoreactive and PR-immunoreactive carcinomatous cell populations when used 2 different type of antibodies, in the ACIS automated method.

Comparative analysis of methods for accurate recognition of cells through nuclei staining of Ki-67 in neuroblastoma and estrogen/progesterone status staining in breast cancer.

OBJECTIVE: To compare 2 automatic systems for the recognition and counting of 2 different families of cells through nuclei staining: Ki-67 in neuroblastoma and estrogen/progesterone (ER/PR) status staining in breast cancer. STUDY DESIGN: Morphology-based segmentation strategies and the Support Vector Machine approach have been used for the accurate extraction and recognition of the cells. To achieve the highest possible accuracy, 2 specialized systems specially suited for Ki-67 and ER/PR staining have been developed. RESULTS: The testing set of histologic slides of Ki-67 and ER/PR staining has been assessed by our system and the results compared to the score of a human expert. The results are in good agreement. The average differences are within the acceptable limits of 10%. The main advantage of the system is its absolute repeatability of scores. CONCLUSION: The proposed computer-assisted automatic system of cell extraction and recognition through nuclei staining has confirmed sufficient accuracy for the tested images and may provide a useful tool for cell recognition and counting on the basis of histologic slides with Ki-67 and ER/PR staining.

Serum matrix metalloproteinases MMP-2 and MMP-9 and metalloproteinase tissue inhibitors TIMP-1 and TIMP-2 in diabetic nephropathy.

BACKGROUND: The regulation of mesangial extracellular matrix (ECM) turnover engages a number of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). High glucose concentration affects ECM degradation and the activities of MMPs and TIMPs. ECM accumulation is involved in the pathogenesis of diabetic nephropathy. METHODS: Serum MMP-9, MMP-2, TIMP-2 and TIMP-1 were measured with ELISA in patients with either chronic renal failure (CRF, n=20), type 2 diabetes mellitus (DM2, n=16) or diabetic nephropathy (DM2+CRF, n=14), and healthy controls (n=20). RESULTS: Diabetic nephropathy was related with profound decrease of serum TIMP-2 (122.2 +/- 47.2 vs. 263.0 +/- 89.2 ng/mL), TIMP-1 (242.5 +/- 96.9 vs. 347.4 +/- 87.2 ng/mL) and MMP-2 (385.4 +/- 42.6 vs. 517.2 +/- 75.4 ng/mL) (p<0.001). Both TIMP-1 and TIMP-2 were reduced in diabetic nephropathy in comparison with either diabetes alone (p<0.01 and p<0.001; respectively) or CRF alone (p<0.001 for both). An approximately 2-fold increase of MMP-9/TIMP-1 and MMP-2/TIMP-2 ratio was found in diabetic nephropathy when compared with diabetes with normal renal function (p<0.01). Further, in DM2 patients, TIMP-2 was decreased when compared with CRF alone (219.2 +/- 71.8 vs. 296.8 +/- 58.4 ng/mL). MMP-2 was lowered in both groups of DM2 and CRF patients (413.8 +/- 59.0 ng/mL and 409.7 +/- 93.1 ng/mL, vs. normal control value of 517.2 +/- 75.4 ng/mL; p<0.001). CONCLUSIONS: These data indicate that circulating TIMP-1, TIMP-2 and MMP-2 are decreased in patients with diabetic nephropathy when compared with either CRF or diabetes.

[Endoscopic surgery in treatment of inverted papilloma nose et paranasal sinuses in the older patient]. / Brodawczak odwrócony nosa i zatok przynosowych leczony chirurgia endoskopowa u chorej w wieku podeszlym.

Inverted papilloma is a benign neoplasm of uncertain ethiology. The tumour is considered locally aggressive, its tendency to invade adjacent paranasal structures and high recurrence rate causes. In this work was described one case of 102 year's old patient with unilateral nasal obstruction since 5 month and headache. Endoscopy surgery was applied. Histological examination revealed inverted papilloma.

[The possibilities and barriers in endoscopic examination and treatment of esophagus cancer]. / Mozliwosci endoskopli w rozpoznawaniu i leczeniu nowotworów przelyku.

When it comes to the effective treatment of esophagus tumor is one of the most difficult tumor to treat of all tumors of alimentary tract. It is due to the biological features of this tumor and lack of its early clinical symptoms, which would allow to recognize it in the early stage and to begin with a radical treatment. The main method to recognize the esophagous tumor is the endoscopic treatment with histopathology, and the endosonography is the best method which allows to estimate its local advancement. Endoscopy allows to detect pre-malignant phase and the early stage as well use of the alternative to surgical endoscopic methods of the treatment. Unfortunately, endoscopy made to patients with swallow discordance usually shows the advanced esophagus tumor, which is to be qualified for palliative treatment that include: widening of esophagus, different methods of ablation, endoscopic stenting and implementing alimentary gastrostomy. The choice of the treatment method should be based on the type of the tumor and the advancement stage of disease, but the clinical practice shows that the method of treatment is determined by the availability of the methods and experience. The parallel use of different methods of treatment may give better results, but the wide range of treatment methods shows the lack of optimal way of treatment, on the other hand gives the possibility to chose the individual method of optimal treatment. Surgical treatment is still method of choice in the esophagus tumor, but it is often impossible to perform or is appears to be ineffective and it creates the possibility of use of alternative endoscopic methods. The article shows the possibilities of diagnosing and different endoscopic techniques for radical treatment of earlyphase and palliative treatment of advanced tumor of esophagus.

[Endoscopic polypectomy in colorectal cancer prophylaxis]. / Endoskopowa polipektomia w profilaktyce raka jelita grubego.

The increasing incidence of colorectal cancer force to find new and enhance old methods of diagnostic methods that could allow the pre-magnant or early cancer lesions diagnosing. Those could be more effectively and with less invasive surgical and endoscopic methods cured. The proposed method in prophylaxis is colonoscopy that allow the lesion detection as well as apply curative method what is better for patient and decrease cost of procedure. Diagnostic efficacy is dependant on the experience of the person performing the procedure. Only in some cases applying methods as inking or magnifying increase diagnostic yield. Early lesion detection and treatment of colorectal adenomas that could be protruding, flat or concave lead to significant decrease of colorectal cancer incidence. Implementing the wider array of indications and mucosectomy to clinical practice improve treatment efficacy but require proper preparation and experience as well as qualification for the procedure. Despite the development of new diagnostic methods basis the proper evaluation of mucosal layer during endoscopy and histopatological assessmnet of found lesions. Colonoscopy is also widely accepted method of colorectal cancer surveillance after endoscopic or surgical procedures in colorectal treatment.

[Pathomorphologic diagnostic guidelines for colon polyps]. / Zasady diagnostyki patomorfologicznej polipów jelita gubego.

Among polypoid laesions of the large bowel the great attention should be payed to Adenomas because of they neoplastic transformation capabilities. In this paper a classification of neoplastic adenomas extended with serrated adenoma and hyperplastic polyps with presence of tubular adenoma structures has been presented. The recommended laboratory procedures used in preparing of endoscopy material has been discussed. The role of Pathologists and Gastrologist in proper diagnosis of precancerous and early cancerous changes has been also discussed.

[Clinico-pathological cooperation rules in biologic material processing]. / Zasady wspólpracy k iniczno-patomorfologicznej w opracowywaniu materialu biologicznego.

The principles of collaboration between a clinician and a pathomorphologist in diagnosis of the intestinal diseases were presented. The optimal criteria of drawing and fixing cytological and tissue samples were presented. The importance of extensive clinical information for pathomorphological diagnosis was proved. It was indicated that an early collaboration between a clinician and a pathomorphologist in the proccess of clinical diagnostic planning is necessary.

[New method of biopsy of palatine tonsils and its role in the indication for tonsillectomy (preliminary report)]. / Nowa metoda oligobiopsji migdalków podniebiennych i jej znaczenie w kwalifikacji do tonsillektomii (doniesienie wstepne)

In our work we presented a new method of chronic tonsillitis diagnosis and evaluated its usefulness in qualification to tonsillectomy. Oligobiopsies were obtained from right and left palatine tonsils of 36 patients who were qualified to tonsillectomy on the basis of past medical history, otolaryngological examination and laboratory tests findings. Following gauge of biopsy instruments were used: 2 mm, 3 mm, 4 mm. The material obtained at biopsies and tonsillectomies was histopathologically evaluated. Histopathological findings of specimens obtained at oligobiopsies and post tonsillectomies were compared. The presence of interstitial fibrosis, neutrophils infiltrates, hyperplastic changes of lymphoid follicles of palatine tonsils were evaluated in oligobiopsy specimens in comparison to the results of postoperative specimen findings. Full concordance of histopathological findings obtained at oligobiopsy and post tonsillectomy was noted with the use of biopsy instruments of 3 mm and 4 mm gauge. Specimens obtained at oligobiopsies had a great diagnostic value. It was proved that the biopsy performed according to our method has a great diagnostic value and helps surgeon to qualify patient to tonsillectomy.

[Rare case of papillary cystic carcinoma of ectopic thyroid gland in glossothyroid ductus]. / Rzadki przypadek raka brodawczakowato- torbielowatego ektopicznego gruczolu tarczowego przewodu tarczowo-jezykowego.

The neck midline cyst is the most common congenital malformation in the neck area. In described case of 33 years women the diagnosis was established on clinical examination, the results of USG and the aspiration biopsy. The tumor was surgically excited and the microscopy examination revealed papillary cystic carcinoma of ectopic thyroid gland in glossothyroid ductus. In the every case of neck tumor we have to consider neoplastic process.

[Possibilities and limitations of morphological diagnosis in early stages of colitis]. / Mozliwosci i ograniczenia diagnostyki patomorfologicznej we wczesnych fazach zmian zapalnych wjelicie grubym.

In the paper the most frequent difficulties in diagnosis of early stages of colitis has been presented. Also the most frequent sources of errors depending on methods of obtaining and fixing of material as well as diagnostic possibilities of various kinds of obtained biopsy material has been shown. The role of close cooperation between clinician and pathologist in the earliest stages of diagnostic efforts and final diagnosis has been emphasized.

[Cancer antigen 125 as a marker of ascites in patients with liver cirrhosis]. / Antygen rakowy CA-125 markerem wodobrzusza w marskosci watroby.

Cancer antigen 125 (CA-125) is usually used to monitor the course of epithelial ovarian cancer. It has recently been reported that liver cirrhosis is associated with elevated serum CA-125, especially in the presence of ascites. The aim of the study was to evaluate CA-125 as a marker of ascites in patients with liver cirrhosis. Seventy-two (72) patients with liver cirrhosis of different aetiology were studied. Ca-125 levels were measured in stored serum collected from the patients. Ca-125 concentrations were elevated in patients with liver cirrhosis and ascites, irrespective of patients' sex and cirrhosis aetiology. CA-125 concentrations were normal in cases of liver cirrhosis without ascites. Elevated cancer antigen 125 is a sensitive marker of cirrhotic ascites. An inappropriate use of this test in cases of liver cirrhosis in females may suggest the ovarian cancer incident and lead to unnecessary surgical intervention.