Testicular Germ Cell Tumor Tissue Biomarker Analysis: A Comparison of Human Protein Atlas and Individual Testicular Germ Cell Tumor Component Immunohistochemistry.

The accurate management of testicular germ cell tumors (TGCTs) depends on identifying the individual histological tumor components. Currently available data on protein expression in TGCTs are limited. The human protein atlas (HPA) is a comprehensive resource presenting the expression and localization of proteins across tissue types and diseases. In this study, we have compared the data from the HPA with our in-house immunohistochemistry on core TGCT diagnostic genes to test reliability and potential biomarker genes. We have compared the protein expression of 15 genes in TGCT patients and non-neoplastic testicles with the data from the HPA. Protein expression was converted into diagnostic positivity. Our study discovered discrepancies in three of the six core TGCT diagnostic genes, POU5F1, KIT and SOX17 in HPA. DPPA3, CALCA and TDGF1 were presented as potential novel TGCT biomarkers. MGMT was confirmed while RASSF1 and PRSS21 were identified as biomarkers of healthy testicular tissue. Finally, SALL4, SOX17, RASSF1 and PRSS21 dysregulation in the surrounding testicular tissue with complete preserved spermatogenesis of TGCT patients was detected, a potential early sign of neoplastic transformation. We highlight the importance of a multidisciplinary collaborative approach to fully understand the protein landscape of human testis and its pathologies.

CORRELATION OF EXPRESSION OF TGF- ß AND MMP2 BETWEEN PROSTATIC ADENOCARCINOMA AND ADJACENT UNAFFECTED PARENCHYMA.

In prostate adenocarcinoma, both tumorous stroma and epithelium have important role in tumor progression. Transforming growth factor beta (TGF- ß) is a promotor in advanced stages of prostate cancer. Matrix Metalloproteinase 2 (MMP2), the endopeptidase that degrades extracellular matrix is considered to be overexpressed in prostatic carcinoma related to its growth and aggressiveness. Therefore, the aim was to analyze the expression of proteins TGF- ß and MMP2 between both epithelium and stroma of prostatic adenocarcinoma and adjacent unaffected parenchyma. The intensity of TGF- ß and MMP2 expression in epithelium, tumorous stroma and adjacent unaffected parenchyma was analyzed in 62 specimens of prostatic adenocarcinoma by microarray-based immunohistochemistry. TGF- ß was more expressed in tumorous than in prostate stroma (p =0.000), while no statistical significance in case of MMP2 (p = 0.097) was found. MMP2 was more expressed in tumorous than in prostate epithelium (p =0.000), while no statistical significance in case of TGF- ß (p = 0.096) was observed. The study results indicate that both tumorous stroma and epithelium have a role in tumor progression and support potential role of TGF- ß and MMP2 in prostatic adenocarcinoma progression.

cfDNA methylation in liquid biopsies as potential testicular seminoma biomarker.

Background: Seminoma is a testicular tumor type, routinely diagnosed after orchidectomy. As cfDNA represents a source of minimally invasive seminoma patient management, this study aimed to investigate whether cfDNA methylation of six genes from liquid biopsies, have potential as novel seminoma biomarkers. Materials & methods: cfDNA methylation from liquid biopsies was assessed by pyrosequencing and compared with healthy volunteers' samples. Results: Detailed analysis revealed specific CpGs as possible seminoma biomarkers, but receiver operating characteristic curve analysis showed modest diagnostic performance. In an analysis of panels of statistically significant CpGs, two DNA methylation panels emerged as potential seminoma screening panels, one in blood CpG8/CpG9/CpG10 (KITLG) and the other in seminal plasma CpG1(MAGEC2)/CpG1(OCT3/4). Conclusion: The presented data promote the development of liquid biopsy epigenetic biomarkers in the screening of seminoma patients.

Seminoma belongs to testicular cancer, which represents a common malignancy among men of reproductive age. Diagnosis of seminoma is a multistep process that also includes checking tumor biomarkers from blood. However, these biomarkers are not specific for seminoma and to conclude a definite diagnosis of seminoma immunohistochemical analysis is needed, which requires the removal of a whole or partial testicle. Therefore, there is a need for novel, noninvasive biomarkers. cfDNA is the most extensively investigated source of minimally invasive tumor markers. Therefore, this study investigated cfDNA methylation of six genes as potential noninvasive biomarkers for the management of seminoma patients. By examining CpG sites of selected genes by pyrosequencing, the authors detected significant differences. However, receiver operating characteristic curve analysis showed modest results. Therefore, the authors tested possible panels of significantly different CpGs and detected two possible DNA methylation panels for seminoma screening. These findings suggest the further investigation of possible epigenetic biomarkers for seminoma patient management from liquid biopsies.

In Search of TGCT Biomarkers: A Comprehensive In Silico and Histopathological Analysis.

Testicular germ cell tumors (TGCTs) are ever more affecting the young male population. Germ cell neoplasia in situ (GCNIS) is the origin of TGCTs, namely, seminomas (SE) and a heterogeneous group of nonseminomas (NS) comprising embryonal carcinoma, teratoma, yolk sac tumor, and choriocarcinoma. Response to the treatment and prognosis, especially of NS, depend on precise diagnosis with a necessity for discovery of new biomarkers. We aimed to perform comprehensive in silico analysis at the DNA, RNA, and protein levels of six prospective (HOXA9, MGMT, CFC1, PRSS21, RASSF1A, and MAGEC2) and six known TGCT biomarkers (OCT4, SOX17, SOX2, SALL4, NANOG, and KIT) and assess its congruence with histopathological analysis in all forms of TGCTs. Cancer Hallmarks Analytics Tool, the Search Tool for the Retrieval of Interacting Genes/Proteins database, and UALCAN, an interactive web resource for analyzing cancer OMICS data, were used. In 108 TGCT and 48 tumor-free testicular samples, the immunoreactivity score (IRS) was calculated. SE showed higher frequency in DNA alteration, while DNA methylation was significantly higher for all prospective biomarkers in NS. In GCNIS, we assessed the clinical positivity of RASSF1 and PRSS21 in 52% and 62% of samples, respectively, in contrast to low or nil positivity in healthy seminiferous tubules, TGTCs as a group, SE, NS, or all NS components. Although present in approximately 80% of healthy seminiferous tubules (HT) and GCNIS, HOXA9 was diagnostically positive in 64% of TGCTs, while it was positive in 82% of NS versus 29% of SE. Results at the DNA, mRNA, and protein levels on putative and already known biomarkers were included in the suggested panels that may prove to be important for better diagnostics of various forms of TGCTs.

PROSTATE-SPECIFIC ANTIGEN (PSA) VALUES IN PATIENTS WITH LOW- AND HIGH-RISK PROSTATIC ADENOCARCINOMA.

Prostatic adenocarcinoma (PC) comprises around 19% of malignancies in Croatian male population. On the basis of PSA value, Gleason score, grading group and clinical stage, PC can be classified into low- and high-risk groups which is significant for different therapeutic regimens and prognostic outcomes. In this retrospective study, we analyzed the difference in preoperative PSA value in a group of 272 patients who underwent radical prostatectomy and were diagnosed with PC adenocarcinoma in our institution in a period from January 1st, 2018 untill December 31st, 2018. Subsequently, they were divided into low- and high-risk prostatic adenocarcinoma groups. Our results demonstrated positive correlation in preoperative PSA values between the groups and therefore support the use of PSA as one of the parameters in defining low- and high-risk prostatic adenocarcinoma categories.

Rare Occurrence of Incidental Finding of Noninvasive Follicular Thyroid Neoplasm With Papillary-Like Nuclear Features in Hürthle Cell Adenoma.

INTRODUCTION: Hürthle cell adenoma is a rare benign lesion of the thyroid gland, however, controversies about its potential malignant behavior still remain. Among thyroid neoplasms, papillary carcinoma is the most common variant with great variety of histological subtypes demonstrating different biological behavior. AIM: To raise the awareness of possible coexistence of these two lesions and discussion about possible therapeutic approaches. CASE REPORT: A 42 year old female patient was examined because of the pain in the thyroid area. Cytological examination suggested Hürthle cell adenoma. Subsequently, right thyroid lobectomy was performed. Intraoperative frozen sections confirmed the diagnosis, yet final histological analysis revealed encapsulated follicular variant of papillary thyroid carcinoma (EFVPTC), now reclassified as noninvasive follicular thyroid neoplasm with papillary- like nuclear features (NIFTP) within the adenoma, which was not noticed through scintigraphy, ultrasound, cytological and frozen section analysis. CONCLUSIONS: Problems concerning both diagnostic and therapeutic approach to these lesions are being discussed, since opinions reported in the literature are divided, posing great challenge for the clinician in determining adequate therapeutic procedures.

Expression of LMO2 in Prostate Carcinoma and Adjacent Prostatic Parenchyma.

LMO2 (LIM domain only) is a member of transcription factor family of proteins characterized by their cysteine-rich, zinc-binding LIM domains. Its expression in prostate cancer cells, as well as in adjacent stroma, is described in a study in a cohort of 83 patients treated with radical prostatectomy for clinically localized prostate adenocarcinoma. Authors found that LMO2 overexpression in prostate cancer was strongly associated with features indicative of worse prognosis (higher preoperative PSA, higher Gleason score, positive surgical margins, and extraprostatic extension of disease). Expression of LMO2 was also associated with biochemical disease progression. We analysed immunohistochemical expression of LMO2 in prostate cancer epithelial and stromal cells, as well as in adjacent parenchyma. Significant negative correlation between glandular expression of LMO2 in carcinoma and stromal expression in BPH (ρ = -0.238, P = 0.033) was found, but also be-tween stromal expression in carcinomas and glandular expression in BPH (ρ = -0.255, P = 0.021). Positive correlation was found between stromal expression in BPH and stromal expression in carci-nomas (ρ = 0.306, P = 0.005). Study results support the potential role of LMO2 in prostatic carcino-genesis and cancer progression.

Expression of plakophilin 3 in diffuse malignant pleural mesothelioma.

Diffuse malignant pleural mesothelioma (DMPM) is the most common primary malignant pleural neoplasm still posing major diagnostic, prognostic and therapeutic challenges. Plakophilins are structural proteins considered to be important for cell stability and adhesion in both tumor and normal tissues. Plakophilin 3 is a protein present in desmosomes of stratified and simple epithelia of normal tissues with presence in malignant cells of various tumors where it participates in the process of tumorigenesis. The aim of this study was to investigate the expression of plakophilin 3 protein in DMPM, but also to study its prognostic significance and relation to histologically accessible parameters of aggressive growth. Archival samples of tissue with established diagnosis of DMPM and samples of normal pleural tissue were used. Tumor samples were classified into three histological types of DMPM (epithelioid, sarcomatoid and biphasic). Additional subclassification of epithelioid mesotheliomas into nine patterns based on the prevalent histological component of the tumor was then performed. After immunohistochemical staining, cytoplasmic and membrane immunopositivity of tumor cells was assesed by scoring the intensity of the staining from 0 (no staining) to 4 (very strong staining). Prognostic value and expression of plakophilin 3 with consideration to histologically estimated aggression in tumor growth were then statistically analyzed using non- parametric tests. The results demonstrated higher level of plakophilin 3 expression in tumor samples with histologically more aggressive tumor growth, but no significant prognostic value. According to our study, plakophilin 3 appears to be involved in tumor invasion in malignant mesothelioma.

Pulmonary myelolipoma containing osseous tissue: An unexpected finding at autopsy.

Pulmonary myelolipoma is a very rare benign tumor composed of mature adipose tissue and hematopoietic elements such as erythroid, myeloid and megakaryocytic. It usually represents accidental finding during autopsy or chest imaging, since most cases are asymptomatic. Larger masses can lead to hemorrhage, chest pain and chest organ compression. We present a case of incidental finding of pulmonary myelolipoma during the autopsy of an 83- year old woman who died of abdominal aortic rupture. In the right lower lung lobe, solitary, well-circumscribed yellow-brown nodule which was 3 cm in its longest diameter was found. Pathohistological analysis revealed tumor composed of mature adipose tissue and hematopoietic cells (myeloid cells, megakaryocytes, erythroid cells) with fragments of mature bone tissue. Differential diagnosis of pulmonary myelolipoma includes lipoma, liposarcoma, hamartoma, phlebangioma, teratoma and extramedullary hematopoiesis. In majority of cases, tumor removal is not necessary, however, larger lesions should be surgically removed. No cases of malignant transformation or recurrence have so far been reported in the literature.

Enterobius vermicularis in the Endometrium of the Uterus: A Case Report.

Enterobius vermicularis is an intestinal nematode of humans and the most common helminth infection. Main transmission path is direct contact between infected and uninfected person meaning ingestion of the eggs. Human infections are usually asymptomatic or manifest as perianal itching. Although ectopic locations are uncommon, Enterobius can occasionally be detected in appendix, kidney, male urinary tract and female genital tract. We present a case from Varazdin General Hospital, Varazdin, Croatia in 2012, involving a 90-yr-old female patient who underwent hysterectomy leading to accidental finding of E. vermicularis in the uterus despite being asymptomatic for enterobiasis. Since there were no signs and symptoms of parasitic infection, no antiparasitic drugs were administered. Parasite was not observed during macroscopic examination, yet microscopic examination of the material demonstrated helminth within endometrium surrounded by dense inflammatory infiltrate, predominantly lymphocytes and some eosinophils. Internal structures of the parasite were collapsed, while well-developed musculature and cuticle were preserved. We present this case to educate and remind physicians on this parasitosis as possible diagnosis. Although non-gastrointestinal locations of Enterobius infestation are rare, this infection should be considered in patients with abdominal pain, genitourinary symptoms, and pelvic pain in order to apply appropriate treatment and prevent further complications.

Colonic Perforation in a Young Tetraplegic Male Caused by Zucchini.

INTRODUCTION: Colonic perforation is a clinical condition which occurs due to variety of reasons, such as intrinsic disorders of the intestine, extrinsic causes, but also due to presence of foreign bodies. Foreign objects enter gastrointestinal tract by oral or transanal introduction. CASE REPORT: we present an uncommon case of a 26- year-old tetraplegic male, whose death was a consequence of a widespread purulent peritonitis provoked by colonic perforation inflicted by an unusual foreign body, transanally introduced 28 centimeters long zucchini (Cucurbita pepo L.). CONCLUSIONS: we share our experience in order to emphasize the importance of consideration and early recognition of foreign body presence in the alimentary tract as possible diagnosis.

Cathepsin D and its prognostic value in neuroepithelial brain tumors.

Expression of Cathepsin D (Cath D) in some primary neuroepithelial brain tumors and its prognostic value were studied. The research included 65 samples of human primary neuroepithelial brain tumors. There were 50 glial tumors (10 diffuse astrocytomas (DA), 15 anaplastic astrocytomas (AA), 25 glioblastomas (GB), 15 embryonic tumors (15 medulloblastomas (MB) as well as 5 samples of normal brain tissue. Immunohistochemical method was applied to monitor diffuse positive reaction in the cytoplasm of brain tumor cells, endothelial cells and tumor stromal cells and showed diffuse positive reaction for Cath D in the cytoplasm of brain tumor cells, endothelial cells and stromal cells in all analyzed samples of DA, AA, GB and MB as well as in microglial cells, neurons and in endothelial cells in all analyzed samples of normal brain tissue. Qualitative analysis of Cath D expression in the cytoplasm of brain tumor cells and endothelial cells as well as the percentage of brain tumor cells, endothelial cells and stromal cells immunopositive for Cath D showed that there was difference between analyzed brain tumor groups, but according to statistical tests the difference was not statistically significant. Survival correlated with the percentage of stromal cells immunopositive for Cath D. Survival prognosis was influenced by the percentage of stromal cells immunopositive for Cath D and tumor grade. The obtained results singled out the percentage of stromal cells immunopositive for Cath D as an independent parameter. The results of this research on the prognostic value of Cath D in some primary brain tumors of neuroepithelial origin indicate that there is real possibility to use Cath D as an independent prognostic factor in human glioma progression and thus open up possibilities for further scientific research.

Benign blue naevus of the lungs.

Blue naevus is a dark blue, gray or black lesion consisted of dermal melanocytes and usually found on face, scalp, or on the dorsum of hands and feet. Two well defined histologic and clinical variants, designated as "common" and "cellular", have been recognised. An unusual case of accidentally detected common blue naevus of the lungs has been reported. The specimen of lung tissue was taken during autopsy of a 62-year old woman who died of myocardial infarction. Microscopic analysis revealed the area containing melanocytes filled with melanin pigment.