Down-regulation of neuronal form of Nitric oxide synthase in the Nurse cell of Trichinella spiralis.

The free radical nitric oxide (NO) and Ca2+ are critical regulators of skeletal muscle exercise performance and fatigue. The major source of NO in skeletal muscle cells is the neuronal form of the enzyme Nitric oxide synthase (nNOS). One of the most peculiar characteristics of the Nurse cell of Trichinella spiralis (T. spiralis) is the complete loss of the contractile capabilities of its derivative striated muscle fiber. The aim of the present study was to clarify the expression of nNOS protein and mRNA in striated muscles during the muscle phase of T. spiralis infection in mice. Muscle tissue samples were collected from mice at days 0, 14, 24, and 35 post infection (d.p.i.). The expression of nNOS was investigated by immunohistochemistry, and the expression levels of mRNA of mouse Nitric oxide synthase 1 (Nos1) by real-time PCR. The presence of nNOS protein was still well observable in the disintegrated sarcoplasm at the early stage of infection. The cytoplasm of the developing and mature Nurse cell showed the absence of this protein. At least at the beginning of the Nurse cell development, Trichinella uses the same repairing process of skeletal muscle cell, induced after any trauma and this corroborates very well our results concerning the nNOS expression on day 14 p.i. At a later stage, however, we could suggest that the down-regulation of nNOS in the Nurse cell of T. spiralis either serves a protective function or is an outcome of the genetic identity of the Nurse cell.

Double Trisomy 16 and 22 Clinically Mimic Partial Hydatidiform Mole in a Case of Subsequent Pregnancy Loss.

A case of double trisomy 16 and 22 in the second pregnancy loss is presented. DNA analyses (short tandem repeats genotyping) of miscarriage specimen was indicated because of ultrasound suspicion of partial hydatidiform mole. After the partial hydatidiform mole exclusion, further DNA analyses focused on the most common aneuploidies causing pregnancy loss, detected double trisomy 16 and 22 in the product of conception. The couple was referred to clinical genetic consultation and normal parental karyotypes were proved. For further explanatory purposes, archived material from the first pregnancy loss was analyzed and trisomy of chromosome 18 was detected. By comparison of allelic profiles of the mother, father, and both losses, the maternal origin of all aneuploidies was proven what can be attributed to frequent meiosis errors, probably due to advanced maternal age (44 years at the first loss and 45 years at the second loss). In conclusion, aneuploidies can mimic partial hydatidiform mole. Genetic analysis is helpful on the one hand to rule out partial hydatidiform mole and on the other hand to identify aneuploidies and in this way to determine the cause of miscarriage.

Retroperitoneal abscess after urgent resection of the stomach in a geriatric patient treated with a surgical extraperitoneal approach - case report.

Retroperitoneal abscess after abdominal and retroperitoneal surgery is a relatively rare but serious complication that most often occurs as a result of a healing disorder in the postoperative period. The incidence is not high, in the literature the cases are mostly reported as case reports with a serious clinical course, high morbidity and mortality. The most important factor of effective treatment, after successful diagnosis by CT examination is rapid evacuation of the abscess and retroperitoneal drainage, in which mini-invasive surgical or radiological drainage dominate as methods of choice. Surgical drainage, burdened by higher morbidity and mortality is considered the last resort after failure of mini-invasive methods. In our case report, we present a case of retroperitoneal abscess, arising as a complication after gastric resection, which was evacuated and drained primarily surgically due to unsuitability for radiological intervention.

When Less Is More - Pipelle Endometrial Sampling for Quantification of Uterine Natural Killer Cells in Patients With Recurrent Implantation Failure or Habitual Abortion.

Despite recent advancements in reproductive medicine, recurrent implantation failure and habitual abortion remain ongoing issues. One of the most important aspects of successful implantation is the intricate immune response and regulation necessary for the acceptance of the hemiallogenic embryo. The most numerous immune cells in the decidua are uterine natural killer cells (uNK). Studies suggest that changes in the uNK count and physiology may be responsible for the aforementioned pathological conditions. Thus, testing for uNK may provide valuable insights into their pathogenesis. The study compared Pipelle endometrial sampling with conventional curettage to find out whether the less invasive Pipelle method is a viable alternative of tissue collection. Tissue samples from 14 patients obtained by both methods were examined. The average size of tissue samples obtained with Pipelle was 17 mm2, samples obtained with curettage had on average 34 mm2. Using immunohistochemical visualization of CD56 (NK cells) and granzyme B antigens (serine protease-expressing activation state of NK cells), it was found that the average total count of CD56 / mm2 was for Pipelle 115 and 120 for curettage, respectively. The study also proved a correlation between granzyme B positivity and identification of NK cells clusters. The results indicated that Pipelle endometrial sampling seems a suitable method of tissue harvesting for the purpose of uNK cells examination. Pipelle endometrial sampling is safe, cost-effective and can be performed on an outpatient basis without the need of anesthesia or analgesia. Several issues remain yet to be solved: how to standardize the subsequent uNK testing, how to interpret the results and finally yet importantly, how to use this knowledge in personalized treatment protocols.

Uterine Natural Killer Cells in the Context of Implantation: Immunohistochemical Analysis of Endometrial Samples from Women with Habitual Abortion and Recurrent Implantation Failure.

Infertility affects approximately 48 million couples globally. Despite the enormous progress of the methods of reproductive medicine that has been made since the first test-tube baby was born in 1978, the implantation rate of day-3 embryos is only around 15-20 % and 30 % of day-5 embryos. Numerous strategies aim to improve implantation rates and prevent repeated implantation failure, however there is no specific general recommendation leading to satisfying results. One of the many risk factors relevant in this regard is the uterine immunological make-up, mainly the uterine Natural Killer (uNK) cells. They orchestrate the overall immune response during implantation by influencing trophoblast invasion and vascular remodeling and throughout pregnancy, uNK cells are also the main immune cells at the maternal-foetal interface. Previously, uNK count has been correlated with various fertility issues including idiopathic reccurent miscarriage. The present study used endometrial samples collected from 256 patients with recurrent implantation failure (RIF), habitual abortion (HA) and idiopathic sterility. Samples were collected between day 19 and 21 of the menstrual cycle mainly by Pipelle endometrial sampling. The samples were fixed in formalin for 24 hours and further processed for immunohistochemistry using anti-CD56 to visualize this antigen marker of uNK cells. Immunohistochemical counting was performed to assess the low, normal, or elevated count of uNK cells. According to the one-way ANOVA test, the age of our patients did not have any influence on the count of uNK cells. With Spearman correlation analysis, we found statistically significant correlation (p-value 0.05) of -0.133 between prior miscarriage and lower uNK cell count. Using the same analysis we found statistically significant correlation (correlation 0.233 with p-value 0.01) between number of uNK cells and activation status. Patients with higher uNK cells were more frequenty diagnosed with endometriosis (p-value 0.05, correlation 0.130). Patients with an immunological factor of sterility (defined by a clinical immunologist) had a lower chance of gravidity (-0.203 with p-value 0.01). Based on our results, we can confirm that there is a correlation between RIF, HA, idiopathic sterility, endometriosis, and immunological factor of sterility (uNK cell count). The true predictive value with regard to fertility outcomes needs to be addressed in future research.

Proapoptotic and antiapoptotic stimuli alternation in myocardial infarction experimental model.

Myocardial infarction is a life-threatening complication of the coronary artery disease - the leading cause of premature death worldwide. The severity of this condition is the result of cellular death following the myocardial ischaemia, which occurs via several mechanism including apoptosis. For the research of this condition, animal models are often employed. We established isoprenaline-induced rat model of myocardial infarction, focusing on the immunohistochemical analysis of the expression of antiapoptotic and proapoptotic proteins BCL-2 and BAX, respectively. Apoptosis (based on BAX-positivity) was activated in cardiac muscle cells within the first day, later on day 8 also in fibroblasts of the forming scar tissue. Antiapoptosis in cardiac muscle cells was weak to moderate on the day 1 and 2, on the day 8 macrophages were strongly positive for BCL-2. The results confirmed that programmed cell death as well as mechanisms of antiapoptosis contribute to the pathogenesis of myocardial infarction. Previous research demonstrated that by experimentally affecting proapoptotic and antiapoptotic signals, it is possible to influence various aspects of myocardial infarction including: infarction size, cardiac remodelling and prognosis of the heart failure. Future research is warranted to fully elucidate the role of this process during myocardial infarction, which will result in refined diagnostic and therapeutic strategies (Tab. 1, Fig. 1, Ref. 21). Keywords: myocardial infarction, isoprenaline, apoptosis, necrosis, BCL-2, BAX.

Complex changes of circadian proteins expression in inflammatory bowel disease.

OBJECTIVES AND BACKGROUND: Recently, a possible role of circadian system in the pathogenesis of various gastrointestinal disorders gained an attention. The association of circadian system with immune system activity and reciprocal connection with intestinal microbiota indicate possible links with inflammatory bowel diseases (IBD). METHODS: The retrospective study provided a semiquantitative immunohistochemical analysis of the expression of 8 core circadian proteins (BMAL1, BMAL2, PER1, PER2, PER3, CLOCK, NPAS2 and TIMELESS) in the epithelial cells of intestinal mucosa in 24 patients with Crohn's disease (CD) and 26 patients with ulcerative colitis (UC). Samples from patients without history of IBD served as the control. The BMAL1 protein expression in intramucosal inflammatory cells was explored as well. RESULTS: The expression of 5 core circadian proteins (BMAL1, PER1, PER3, TIMELESS and NAPS2) was decreased in mucosal epithelium of patients with IBD in comparison with the control samples, whereas the expression of BMAL1 and PER1 was more noticeably decreased in UC patients and PER3, TIMELESS and NPAS2 in CD patients. There was a decreased BMAL1 expression in intramucosal inflammatory cells of IBD patients. CONCLUSION: Decreased core circadian proteins expression in colonic mucosa and in intramucosal inflammatory cells of IBD patients indicated a circadian rhythm deregulation as contributing factor in the development of IBD. To our knowledge, this is so far the most extensive immunohistochemical analysis performed on the samples of IBD patients evaluating the changes in circadian protein expression in the intestinal mucosa (Tab. 1, Fig. 2, Ref. 31).

INTRAOCULAR LYMPHOMA WITH RETROBULBAR INFILTRATION. A CASE REPORT.

Subjective and objective symptoms following intraocular lymphoma could lead to a misdiagnosis at the beginning of the disease, which is the cause for the delay in an effective treatment. The most common manifestation of lymphoma is an inflammatory disease affecting the uvea. A multidisciplinary approach to the diagnosis is required. Suspicion based on the ophthalmological examinations has to be verified by histology. We present a case report of a 78-year-old patient examined at our clinic, with progressive loss of vision in the left eye over 6 months, suspected of retinal detachment. Objectively the visual acuity was counting fingers in front of the left eye. Intraocular pressure changed from normotensive to hypertensive values during regular examinations. We realised imaging exams, ultrasonography and magnetic resonance, which proved an intraocular tumour with retrobulbar infiltration and retinal detachment. The patient was indicated for enucleation, which enabled assignment of a histological type of intraocular B-Non-Hodgkin lymphoma from marginal zone B-cells. We sent the patient to a haematologist-oncologist for management of the subsequent treatment and we prescribed an individual prosthesis to the patient after the enucleation. The patient remains under observation; no surgical treatment, chemotherapy or radiotherapy have been used for 15 months after the enucleation. The enucleation was both a diagnostic and treatment modality.

Primary Intrabulbar Neurofibroma.

PURPOSE: Eye globe enucleation due to other than a malignant tumor is very rare today. Solitary intraocular neurofibroma without other signs of neurofibromatosis is a rare benign tumor and few cases have been reported to date. MATERIAL AND METHODS: In 10 year interval from Jan 1 2007 to Dec 31 2016 we analyzed non-malignant eye globe enucleations. RESULTS: Of the 49 enucleated blind eyes, each patient had visual acuity with no light perception, 34 (69.4%) were indicated for enucleation due to complications following previous postoperative surgery after trauma, 14 patients (28.6%) were due to secondary glaucoma and other complications following previous intraocular surgery, and in one patient (2%) the primary isolated intraocular neurofibroma was verified after enucleation. CASE REPORT: A patient with isolated intrabulbar neurofibroma has been monitored since childhood for intraocular lesion and histologically verified at adult age. At the time of enucleation, he was 25 years old, squint since childhood and was observed for hamartoma in his right eye since he was 13 years old. Due to the progression of intrabulbar lesion, loss of visual acuity (functional state - no light perception) and secondary glaucoma, the right eye globe was enucleated at adult age and histopathological examination confirmed intraocular neurofibroma in the absence of neurofibromatosis. CONCLUSION: Every enucleated eye globe should be subjected to a thorough histopathological examination. Isolated intraocular neurofibromas can occur as isolated orbital or intrabulbar masses without systemic features.

Enucleation after Stereotactic Radiosurgery in Patients with Uveal Melanoma.

INTRODUCTION: In the past enucleation was the treatment of choice for all the patients with uveal melanoma. Nowadays, we prefer glope-sparing treatment modalities, except for large tumors, tumors with extrascleral extension and painful blind eyes. Most of the patients perform radiotherapy or local resection techniques. In Slovak Republic, the only one possibility is a stereotactic radiotherapy on a linear accelerator LINAC. Nevertheless, enucleation after radiotherapy is necessary for some patients. The causes are postradiation complications, mainly neovascular glaucoma, tumor recurrence, tumor progression or patient´s decision. MATERIAL AND METHODS: The retrospective non-randomised study of 168 eyes of the patients with choroidal or ciliary body melanoma, who performed one-day session stereotactic radiosurgery at the linear accelerator LINAC during period 2007-2016. The data about postradiation complications were analysed based on the medical records of the patients and the data about enucleated eyes based on the histopathological findings. RESULTS: The occurence of enucleation after radiotherapy in our cohort was 17 % (28 patients), with median time period after radiotherapy 21,5 months. The most common cause was neovascular glaucoma (82 %), then tumor progression (14 %) and patient´s decision (4 %). The most common histopathological finding was spindle-cell melanoma. DISCUSSION: Others authors describe similar enucleation rate and causes. The histopathological findings indentified more viable melanoma cells in eyes enucleated for tumor progression in comparison with enucleation for other causes. Enucleation may be more difficult and the occurence of postoperative complications can be higher in the eyes after radiotherapy rather than primary enucleation. CONCLUSION: The management of the patients with uveal melanoma is difficult, and requires the cooperation of ophthalmologist, oncologist, radiation physicist and pathologist. Even if we make effort to preserve the eye, enucleation after radiotherapy is necessary in some patients.

Optic nerve orbital meningioma.

INTRODUCTION: Orbital meningioma treatment has achieved significant success over the last period. Primary optic nerve sheath meningiomas by the clinically progressive finding, but still persistent sufficient visual acuity, can be treated by fractionated stereotactic radiotherapy. Surgery is indicated for secondary meningiomas of the orbit. Surgical treatment is indicated due to intracranial tumor propagation of meningioma from the chiasm towards the orbit. MATERIAL AND METHODS: In the period 2014 - 2016 we monitored in dispensary 15 patients with meningioma of the orbit, who were checked at least in yearly intervals and underwent magnetic resonance examination of the orbit and brain. RESULTS: In group of 15 patients with histologically unverified meningioma of the orbit, the surgical solution was indicated in 3 patients. In 14 (93.3%) patients meningiomas were secondary infiltrating orbit from the intracranial part of visual pathways, and in 1 case meningioma was primary arising from the optic nerv. We indicated enucleation and partial exenteration in 3 (20%) patients. Histopathological examination confirmed meningioma - in two cases gr. I., in one patient gr. II. In all of them, more than 5 years after the primary diagnose of the process, the tumor infiltration from the chiasm towards the orbit was the indication for surgery. In one patient with meningioma gr. II in 12 months interval after surgery - exenteration with lid sparing technique, there was a further progression from the sella turcica area and the growth of tumor masses to the area of the orbital conus. Patient underwent secondary surgical reduction of tumor mass of the orbit and treatment with sandostatin. CONCLUSION: When deciding to treat meningioma, it is necessary to involve multidisciplinary collaboration. Ophthalmology examination is important because further treatment is indicated on the basis of changes in visual function in correlation with the imaging methods. In cases of progression of the tumor with the infiltration of the orbit, resulting in the loss of visual acuity, in certain conditions a radical solution - enucleation with partial exenteration of the orbit, is necessary. Key words: meningioma of the orbit, primary tumors of the orbit, secondary tumors of the orbit, enucleation, exenteration with lid sparing technique.

Uveal melanoma survival rates after single dose stereotactic radiosurgery.

This paper presents the long-term results of single dose stereotactic radiosurgery for intraocular uveal malignant melanoma, and summarizes the results of the retrospective study in 170 Slovak patients. A group of uveal melanoma patients (149 choroidal melanoma, 21 ciliary body melanoma) from 20 to 92 years of age with 59 year median were treated in 2001-2016. There were 81males (47.7%) and 89 females 89 (52.3%). The median overall follow-up time was three years. The median tumor volume at baseline was 0.5 cm³ (ranging from 0.2 to 1.6 cm³). The therapeutic dose was 35.0 Gy by 99% of dose volume histogram. The survival after single dose stereotactic radiosurgery was 96% in one year, 93% in two years, 84% in five years, 80% in seven years and 52% in eleven years. Secondary enucleation was necessary for 22 patients because of secondary glaucoma complication. The enucleation free interval ranged from one to six years. The survival rates in five year intervals and necessity of secondary enucleation due to complications after single dose stereotactic radiosurgery is comparable to other techniques.

Conjuctival lesions - the relationship of papillomas and squamous cell carcinoma to HPV infection.

The etiology of papilloma formation is multifactorial. There is a strong association between human papillomavirus (HPV) and the development of these conjunctival lesions. HPV is tumorigenic and commonly produces benign tumors with low malignant potential. Papillomas rarely go through malignant transformation. MATERIAL AND METHODS: Retrospective study of patients with a diagnosis of conjunctival papilloma or squamous cell carcinoma. RESULTS: From a collection of 125 patients with conjunctival non-pigmented tumours in the period from 2007 to 2017, in 119 (95.2%) patients histological examination confirmed papilloma and in 6 (4.8%) patients it confirmed carcinoma. Of the total number of patients, 39 were women (31.2%) and 86 men (68.8%). The mean age of patients was 68.4 years (range 20-94 years). Localization of lesions: bulbar conjunctiva - 65 (52.0%), upper eyelid tarsal conjunctiva + fornix - 6 (4.8%), lower eyelid + fornix - 27 (21.6%), caruncle - 20 (16.0%) and plica semilunaris - 7 (5.6%) patients. In the patient cohort we recorded 2 papillomas that were transformed into squamous cell carcinoma. HPV16 was positive in these patients, the carcinomas were from the area of the bulbar conjunctiva, and the surgical solution was associated with the perioperative administration of Mitomycin C. In one case, the inverted papilloma developed into orbital carcinoma within 2 years of primary excision, and the patient underwent radical surgical procedure (partial exenteration of the orbit) followed by radiotherapy. CONCLUSION: Transformation of the papilloma into the carcinoma is rare, but it must always be taken into consideration in case of a recurrence of the disease. HPV can infect the conjunctiva. The ophthalmologist, in collaboration with a pathologist, may recommend appropriate laboratory tests to confirm the diagnosis. Long-term outpatient follow-up of patients after excision of the conjunctival papilloma is also necessary. Key words: epibulbar tumors, conjunctival tumors, papilloma, carcinoma.

Development of a new method for the preparation of an acellular allodermis, quality control and cytotoxicity testing.

Demand for use of acellular allodermis is high but commercially appropriate products are not used routinely because of very high price and limited availability. These facts did motivate us to prepare acellular allodermis using a new, simple and less expensive method. We have developed a original method for preparation of acellular allogeneic dermis based on action of a proteolytic enzyme in combination with distilled water. Hypotonic environment in comparison with SDS or Triton ansure no toxicity of the final product. Trials for determination of optimal trypsin concentrations, temperature and time of action were performed. According to our results, the use of 2.5% trypsin/EDTA solution overnight at +4 °C was proving to be optimal. The histology confirmed absence of cells in the prepared dermis. No toxicity of final acellular dermis was confirmed by three independent tests (agar diffusion test contact cytotoxicity test and grow curve). The prepared acellular dermis seems to be suitable not only for direct clinical use, but it can be used as a scaffold for cell cultivation as well.

Prognostic value of apoptosis inducing factor in uveal melanoma.

In malignant tumors including uveal melanoma there is a continuous effort in search for additional and relevant factors with predictive value and possible therapeutic indications. In the present work we evaluated the 5-year mortality in a group of patients with surgically treated uveal melanoma and its relation to selected demographic, clinical and histopathological parameters, including the expression of apoptosis inducing factor (AIF) in the neoplastic tissue.We analyzed retrospectively the clinical data of patients with uveal melanoma treated surgically (enucleation, endoresection, exenteration) in the period from 2001 to 2007 (n=54). Immunohistochemical detection of AIF expression in formalin fixed and in paraffin embedded tissue samples was evaluated semiquantitatively, intensity and percentage multiplicative Quick Score (QS) was calculated and compared between patients with over 5 year (n=32) and less than 5 year (n=22) survival. In the analyzed group of 54 patients the 5 year mortality was 41 %. We confirmed the negative prognostic significance of some of the known prognostic factors as the tumor size and volume, T3 and T4 stage in the TNM classification and the mixed histological type of the tumor. Immunohistochemistry performed on 49 melanoma specimens showed AIF cytoplasmic positivity, no nuclear translocation was detected. The cut-off value of AIF expression QS ≥ 4 (18) in tumor cells separated the 5 year survival of patients (P = 0.018), odds ratio 5.2 (1.24 - 21.73). Moderate and strong expression of AIF in tumor cells also correlated with less favorable prognosis. Confocal microscopy proved colocalization of AIF with mitochondrial marker in neoplastic cells.The prognosis of patients with uveal melanoma can be more accurate with inclusion of immunohistochemical detection of AIF expression. Increased expression of the AIF protein appears as a new negative prognostic factor predicting the 5 year survival.

[THE BRAF MUTATION AND THE POSSIBILITIES OF UVEAL MELANOMA METASTASING PROGNOSTIC MARKERS IDENTIFICATION]. / MUTÁCIA BRAF A MOZNOSTI IDENTIFIKÁCIE PROGNOSTICKÝCH MARKEROV METASTÁZOVANIA UVEÁLNEHO MELANÓMU.

AIM: The aim is to assess the BRAF gene mutations in patients with posterior uveal melanoma. MATERIAL AND METHODS: Retrospective analysis of the group of patients with malignant melanoma of the uvea, who were indicated to enucleation between 1.1 2015 to 1.3.2016. We analyzed stage of uveal melanoma, volume, cell type and BRAF gene mutations. RESULTS: In clinical study of 20 patients after enucleation due to uveal melanoma at the Department of Ophthalmology in Bratislava, patient age was ranged from 22 to 89 years with a median of 62 years. In 14 patients (70 %) enucleation was the primary treatment and in 6 patients (30 %) enucleation was after irradiation (brachytherapy, Leksell gama knife, linear accelerator). In 17 cases (85 %) the mutation of the BRAF gene was negative and in 3 cases the sample was not assessable for the BRAF mutation. CONCLUSION: BRAF gene mutation is confirmed by several studies found in malignant melanoma of the skin. The histopathology findings in our group did not confirmed our theory, that since the uveal melanoma itself has the similar origin as skin melanoma, should also contain a BRAF mutation.Key words: malignant melanoma of the uvea, mutation of the BRAF gene, chromosomal abnormalities as a prognostic factor.

Fibrillin-1 (FBN-1) a new marker of germ cell neoplasia in situ.

BACKGROUND: Germ cell neoplasia in situ (GCNIS), is preinvasive stage of testicular germ cell tumours (TGCTs). Fibrillins, which are integral components of microfibrils are suggested to be involved in cancer pathogenesis and maintenance of embryonic stem cells pluripotency. The aim of this study was to examine fibrillin-1 (FBN-1) expression in TGCTs patients. METHODS: Surgical specimens from 203 patients with TGCTs were included into the translational study. FBN-1 expression was evaluated in the tumour tissue, in GCNIS and in adjacent non-neoplastic testicular tissue in all available cases. Tissue samples were processed by the tissue microarray method. FBN-1 was detected by immunohistochemistry using goat polyclonal antibody and the expression was evaluated by the multiplicative quickscore (QS). RESULTS: The highest FBN-1 positivity was detected in GCNIS (mean QS = 11.30), with overexpression of FBN-1 (QS >9) in the majority (77.1 %) of cases. Expression of FBN-1 in all subtypes of TGCTs was significantly lower in comparison to expression in GCNIS (all p <0.001). Seminoma had significantly higher expression compared to EC, ChC and TER (all p <0.05), but not to YST (p = 0.84). In non-neoplastic testicular tissue the FBN-1 positivity was very low (mean QS = 0.02). Sensitivity, specificity, positive and negative predictive value of FBN-1 expression for diagnosis of GCNIS were 97.1, 98.8, 98.6 and 97.7 %. CONCLUSIONS: FBN-1 is overexpressed in TGCTs and especially in GCNIS when compared to non-neoplastic testicular tissue in patients with germ cell tumors and could be involved in germ cell neoplasia in situ development.

Dominant neurologic symptomatology in intravascular large B-cell lymphoma.

Intravascular large B-cell lymphoma (IVLBCL) is a rare variant of extranodal large B-cell lymphoma and it is characterized by selective intravascular proliferation of malignant cells. Typical features of the disease include aggressive behavior, rapid and frequently fatal course. Clinical picture is non-specific and heterogeneous, depending on the affected organ. It is not uncommon that this unique type of lymphoma is diagnosed post mortem. Herein, we report two cases of IVLBCL with neurologic symptomatology. In our clinical study patient 1 was an 80-year-old male with mixed paraparesis of lower extremities and difficulties with sphincter control. Patient 2 (56-year-old male) had vision malfunction, mental status changes and defect in phatic and motor functions. In both cases definite diagnosis was established by histological examination of necroptic material. We propose to include IVLBCL in differential diagnostic considerations in patients presenting with gradually impairing neurological status and spinal cord damage of unknown etiology (Fig. 2, Ref. 9).

Prognostic value of programmed-death-1 receptor (PD-1) and its ligand 1 (PD-L1) in testicular germ cell tumors.

BACKGROUND: Testicular germ cell tumors (TGCTs) belong to the most chemosensitive solid tumors; however, a small proportion of patients fail to be cured with cisplatin-based chemotherapy. Inhibitors of PD-1/PD-L1 pathways represent a new class of promising drugs in anticancer therapy. The aim of this study was to evaluate expression and prognostic value of PD-1 and PD-L1 in TGCTs. PATIENTS AND METHODS: Surgical specimens from 140 patients with TGCTs (131 with primary testicular tumor and 9 with extragonadal GCTs) were included into the translational study. PD-1 and PD-L1 expression was detected in the tumor tissue by immunohistochemistry using monoclonal antibodies, scored by the multiplicative quickscore (QS) method, compared with their expression in normal testicular tissue and correlated with clinicopathological characteristics and clinical outcome. RESULTS: None of the GCTs exhibited PD-1 protein, although expression of PD-L1 was significantly higher in GCTs in comparison with normal testicular tissue (mean QS = 5.29 versus 0.32, P < 0.0001). Choriocarcinomas exhibit the highest level of PD-L1 with decreasing positivity in embryonal carcinoma, teratoma, yolk sac tumor and seminoma. PD-L1 expression was associated with poor prognostic features, including ≥3 metastatic sites, increased serum tumor markers and/or non-pulmonary visceral metastases. Patients with low PD-L1 expression had significantly better progression-free survival [hazard ratio (HR) = 0.40, 95% confidence interval (CI) 0.16-1.01, P = 0.008] and overall survival (HR = 0.43, 95% CI 0.15-1.23, P = 0.040) compared with patients with high PD-L1 expression. CONCLUSIONS: In this translational study, we showed, for the first time, the prognostic value of PD-L1 expression in TGCTs and our data imply that the PD-1/PD-L1 pathway could be a novel therapeutic target in TGCTs.

Comparison of structural changes in skin and amnion tissue grafts for transplantation induced by gamma and electron beam irradiation for sterilization.

Sterilization is an important step in the preparation of biological material for transplantation. The aim of the study is to compare morphological changes in three types of biological tissues induced by different doses of gamma and electron beam radiation. Frozen biological tissues (porcine skin xenografts, human skin allografts and human amnion) were irradiated with different doses of gamma rays (12.5, 25, 35, 50 kGy) and electron beam (15, 25, 50 kGy). Not irradiated specimens served as controls. The tissue samples were then thawn and fixed in 10 % formalin, processed by routine paraffin technique and stained with hematoxylin and eosin, alcian blue at pH 2.5, orcein, periodic acid Schiff reaction, phosphotungstic acid hematoxylin, Sirius red and silver impregnation. The staining with hematoxylin and eosin showed vacuolar cytoplasmic changes of epidermal cells mainly in the samples of xenografts irradiated by the lowest doses of gamma and electron beam radiation. The staining with orcein revealed damage of fine elastic fibers in the xenograft dermis at the dose of 25 kGy of both radiation types. Disintegration of epithelial basement membrane, especially in the xenografts, was induced by the dose of 15 kGy of electron beam radiation. The silver impregnation disclosed nuclear chromatin condensation mainly in human amnion at the lowest doses of both radiation types and disintegration of the fine collagen fibers in the papillary dermis induced by the lowest dose of electron beam and by the higher doses of gamma radiation. Irradiation by both, gamma rays and the electron beam, causes similar changes on cells and extracellular matrix, with significant damage of the basement membrane and of the fine and elastic and collagen fibers in the papillary dermis, the last caused already by low dose electron beam radiation.