SFRP4 protein expression is reduced in high grade astrocytomas which is not caused by the methylation of its promoter.

Introduction: Epigenetics play a vital role in stratifying CNS tumors and gliomas. The importance of studying Secreted frizzled-related protein 4 (SFRP4) in gliomas is to improve diffuse glioma methylation profiling. Here we examined the methylation status of SFRP4 promoter and the level of its protein expression in diffuse gliomas WHO grades 2-4. Methods: SFRP4 expression was detected by immunohistochemistry and evaluated semi-quantitatively. In the tumor hot-spot area, the intensity of protein expression in 200 cells was determined using ImageJ (National Institutes of Health, United States). The assessment of immunopositivity was based on the IRS score (Immunoreactivity Score). Promoter methylation was examined by methylation specific-PCR (MSP) in fifty-one diffuse glioma samples and appropriate controls. Isolated DNA was treated with bisulfite conversion and afterwards used for MSP. Public databases (cBioPortal, COSMIC and LOVD) were searched to corroborate the results. Results and discussion: SFRP4 protein expression in glioblastomas was very weak or non-existent in 86.7% of samples, moderate in 13.3%, while strong expression was not observed. The increase in astrocytoma grade resulted in SFRP4 protein decrease (p = 0.008), indicating the loss of its antagonistic role in Wnt signaling. Promoter methylation of SFRP4 gene was found in 16.3% of cases. Astrocytomas grade 2 had significantly more methylated cases compared to grade 3 astrocytomas (p = 0.004) and glioblastomas (p < 0.001), which may indicate temporal niche of methylation in grade 2. Furthermore, the expression levels of SFRP4 were high in samples with methylated SFRP4 promoter and low or missing in unmethylated cases (Pearson's R = -0.413; p = 0.003). We also investigated the association of SFRP4 changes to key Wnt regulators GSK3ß and DKK3 and established a positive correlation between methylations of SFRP4 and GSK3ß (Pearson's R = 0.323; p = 0.03). Furthermore, SFRP4 expression was correlated to unmethylated DKK3 (Chi square = 7.254; p = 0.027) indication that Wnt signaling antagonist is associated to negative regulator's demethylation. Conclusion: The study contributes to the recognition of the significance of epigenetic changes in diffuse glioma indicating that restoring SFRP4 protein holds potential as therapeutic avenue. Reduced expression of SFRP4 in glioblastomas, not following promoter methylation pattern, suggests another mechanism, possible global methylation, that turns off SFRP4 expression in higher grades.

Intraneural Nodular Fasciitis of the Dorsal Scapular Nerve: Case Report and Review of the Literature.

BACKGROUND: Nodular fasciitis is a benign neoplasm occurring predominantly in the subcutaneous tissue. There have been nine intraneural occurrences described in the literature. CASE REPORT: A 37-year-old woman presented with numbness and tenderness in her left shoulder and scapula and a slightly dropped left shoulder, without history of trauma. A magnetic resonance imaging (MRI) of the cervical spine showed a well-circumscribed oval mass deep to the levator scapula muscle. Due to persisting symptoms and an unknown nature of the process, surgical excision was performed, and histopathologic analysis confirmed diagnosis of a benign fibroblastic/myofibroblastic neoplasm, nodular fasciitis. The postoperative course was uneventful and the patient was without symptoms at 4 months of follow-up. METHODS: We reviewed the available literature (PubMed, Google Scholar), with nine published cases of intraneural nodular fasciitis. The reported clinical, radiologic, and histopathologic parameters were evaluated and compared. DISCUSSION: Most of the cases reported in the literature were symptomatic, with tenderness and palpability being the main symptoms. Six of the reported cases occurred in the forearm, whereas three were in the leg. To the best of our knowledge, ours is the first reported case of nodular fasciitis occurring in the trunk. Ours is the only case to display desmin positivity, which supports the reactive hypothesis of nodular fasciitis. CONCLUSION: Intraneural nodular fasciitis is an extremely rare diagnosis. Due to its benign natural course, a multidisciplinary approach with this extremely rare diagnosis in mind is needed to avoid overtreatment.

Endoscopic Endonasal Surgery for a Pterygopalatine Fossa Hydatid Cyst.

Background: Cystic echinococcosis is a manifestation of a zoonosis caused by larvae of the tapeworm Echinococcus granulosus sensu lato and pterygopalatine fossa cases are extremely rare. Clinical Presentation and Findings: A 45-year-old Caucasian female with a history of repeated surgeries for HC was referred to our center for treatment of a cystic mass of the pterygopalatine fossa. Multiorgan dissemination was noted on preoperative imaging. Interventions: An endonasal endoscopic procedure was carried over under general anesthesia and the CE completely removed. Etiology was confirmed by molecular diagnostics. Three weeks after the skull base procedure, the patient underwent a combined abdominal/urological procedure for treatment of other cysts. Conclusion: This case shows that the pterygopalatine fossa HC are amenable to surgical treatment using the endonasal endoscopic approach. Extensive preoperative workup is essential to assess the extent of the disease.

Bilateral Meningioma: A Case Report and Review of the Literature.

Here, we present a rarely seen example of bilateral meningiomas exhibiting different malignancy grades, I (meningothelial) and II (atypical), recorded in a 72-year-old patient. The presence of two separated lesions of different grades in a single patient can elucidate meningioma progression. To this end, the involvement of specific protein markers of epithelial to mesenchymal transition (EMT), the process responsible for progression, was tested in both tumors. Protein expression status of specific epithelial (E-cadherin) and mesenchymal markers (N-cadherin, SNAIL&SLUG and TWIST1) was investigated. Furthermore, markers that are connected to Wnt signaling pathway-beta-catenin, GSK3beta and DVL1-were also analyzed. For signs of neurofibromatosis and schwanomatosis genetic testing was performed. Immunohistochemistry evaluated by immunoreactivity score (IRS) was used to determine the signal strengths and proteins' location. Our results indicated that, in comparison to the grade I tumor, mesenchymal markers SNAIL and SLUG were upregulated in the atypical meningioma. TWIST1, beta-catenin and GSK3beta were upregulated in both grades, while E-cadherin was partially lost. A pronounced cadherin switch could not be established; however, N-cadherin showed widespread tissue presence. Genetic testing did not detect changes of NF2 or SMARCB1 genes denying germline origin of the lesions. The rare presence of two different grades in one patient elucidate previously unknown molecules involved in meningioma progression.

Methylation Patterns of DKK1, DKK3 and GSK3ß Are Accompanied with Different Expression Levels in Human Astrocytoma.

In the present study, we investigated genetic and epigenetic changes and protein expression levels of negative regulators of Wnt signaling, DKK1, DKK3, and APC as well as glycogen synthase kinase 3 (GSK3ß) and ß-catenin in 64 human astrocytomas of grades II-IV. Methylation-specific PCR revealed promoter methylation of DKK1, DKK3, and GSK3ß in 38%, 43%, and 18% of samples, respectively. Grade IV comprised the lowest number of methylated GSK3ß cases and highest of DKK3. Evaluation of the immunostaining using H-score was performed for ß-catenin, both total and unphosphorylated (active) forms. Additionally, active (pY216) and inactive (pS9) forms of GSK3ß protein were also analyzed. Spearman's correlation confirmed the prevalence of ß-catenin's active form (rs = 0.634, p < 0.001) in astrocytoma tumor cells. The Wilcoxon test revealed that astrocytoma with higher levels of the active pGSK3ß-Y216 form had lower expression levels of its inactive form (p < 0.0001, Z = -5.332). Changes in APC's exon 11 were observed in 44.44% of samples by PCR/RFLP. Astrocytomas with changes of APC had higher H-score values of total ß-catenin compared to the group without genetic changes (t = -2.264, p = 0.038). Furthermore, a positive correlation between samples with methylated DKK3 promoter and the expression of active pGSK3ß-Y216 (rs = 0.356, p = 0.011) was established. Our results emphasize the importance of methylation for the regulation of Wnt signaling. Large deletions of the APC gene associated with increased ß-catenin levels, together with oncogenic effects of both ß-catenin and GSK3ß, are clearly involved in astrocytoma evolution. Our findings contribute to a better understanding of the etiology of gliomas. Further studies should elucidate the clinical and therapeutic relevance of the observed molecular alterations.

Adrenal Cortical Adenoma in the Spinal Canal: A Case Report and Review of the Literature.

Ectopic adrenal cortical neoplasms of the spinal cord are extremely rare. To date only 10 such cases have been described. We present a case of a 46-year-old woman with lower back pain radiating to the right gluteal and posterior femoral regions, without a history of traumatic injury. Magnetic resonance imaging (MRI) of the thoracic and lumbar spine showed an intradural, extramedullary, well-circumscribed, contrast-enhancing lesion located in the T12-L1 region, hypo- to isointense on T2-weighted imaging, and isointense on T1. Complete surgical removal of the lesion, measuring 3 × 2.5 × 1 cm, was performed. The histopathologic findings revealed the lesion was an ectopic adrenal cortical adenoma, with sheets and nests of round and polygonal cells, mostly round regular nuclei, abundant eosinophilic cytoplasm, 1 mitosis per 10 high-power fields, and without necrosis. These tumors have nonspecific MRI features and therefore can be easily confused with other common spinal tumor types such as ependymoma, schwannoma, meningioma, and metastasis. Although rare, ectopic adrenal spinal cord adenomas should be taken into account in the differential diagnosis of spinal canal intradural neoplasms.

Polymethylmethacrylate cranioplasty using low-cost customised 3D printed moulds for cranial defects - a single Centre experience: technical note.

We report our experience with 3D customised cranioplasties for large cranial defects. They were made by casting bone cement in custom made moulds at the time of surgery. Between October 2015 and January 2018, 29 patients underwent the procedure; 25 underwent elective cranioplasties for large cranial defects and four were bone tumour resection and reconstruction cases. The majority of patients (96.5%) reported a satisfactory aesthetic outcome. No infections related to the surgical procedure were observed in the follow-up period. The method proved to be effective and affordable.

Different behaviour of DVL1, DVL2, DVL3 in astrocytoma malignancy grades and their association to TCF1 and LEF1 upregulation.

Key regulators of the Wnt signalling, DVL1, DVL2 and DVL3, in astrocytomas of different malignancy grades were investigated. Markers for DVL1, DVL2 and DVL3 were used to detect microsatellite instability (MSI) and gross deletions (LOH), while immunohistochemistry and immunoreactivity score were used to determine the signal strengths of the three DVL proteins and transcription factors of the pathway, TCF1 and LEF1. Our findings demonstrated that MSI at all three DVL loci was constantly found across tumour grades with the highest number in grade II (P = 0.008). Collectively, LOHs were more frequent in high-grade tumours than in low grade ones. LOHs of DVL3 gene were significantly associated with grade IV tumours (P = 0.007). The results on protein expressions indicated that high-grade tumours expressed less DVL1 protein as compared with low grade ones. A significant negative correlation was established between DVL1 expression and malignancy grades (P < 0.001). The expression of DVL2 protein was found similar across grades, while DVL3 expression significantly increased with malignancy grades (P < 0.001). The signal strengths of expressed DVL1 and DVL3 were negatively correlated (P = 0.002). However, TCF1 and LEF1 were both significantly upregulated and increasing with astrocytoma grades (P = 0.001). A positive correlation was established between DVL3 and both TCF1 (P = 0.020) and LEF1 (P = 0.006) suggesting their joint involvement in malignant progression. Our findings suggest that DVL1 and DVL2 may be involved during early stages of the disease, while DVL3 may have a role in later phases and together with TCF1 and LEF1 promotes the activation of Wnt signalling.

Hypermethylation of Secreted Frizzled Related Protein 1 gene promoter in different astrocytoma grades.

AIM: To identify the involvement of Secreted Frizzled Related Protein 1 (SFRP1) promoter hypermethylation in different malignancy grades of astrocytoma and assess its association with beta-catenin, lymphoid-enhancer factor 1, and T-cell factor 1. METHODS: Twenty-six astrocytoma samples were collected from 2008-2015. Promoter hypermethylation was evaluated by methylation-specific polymerase-chain-reaction and protein expression by immunohistochemistry and stereological analysis. The staining intensity was scored by comparing immunoreactivity with normal tissue and by using 10% and 50% cut-offs. RESULTS: SFRP1 promoter methylation was found in 32% of astrocytomas. The number of hypermethylated samples increased in higher astrocytoma grades and was the highest in glioblastoma (P=0.042 compared to other astrocytoma grades). There was 45.8% of samples with the lack of or weak expression of SFRP1 protein and 29.2% with strong expression. Samples with methylated promoter expressed significantly less SFRP1 than samples with unmethylated promoter (P=0.031). Beta-catenin expression levels were elevated. Yet, glioblastomas with unmethylated SFRP1 promoter had significantly less beta-catenin (P=0.033). Strong expression of lymphoid-enhancer factor 1 was associated to higher astrocytoma grades (P=0.006). CONCLUSION: SFRP1 gene was epigenetically silenced in glioblastomas when compared to low astrocytoma grades, which may suggest that the lack of its protein is involved in astrocytoma progression.

Intramedullary Spinal Cord Lipoma Mimicking a Late Subacute Hematoma.

Spinal cord lipomas are rare and benign tumors which may cause progressive neurological deficits due to their local expansion. We present the case of a 59-year-old male patient with severe lumbosacral pain and slowly progressive paresis of the right leg, misdiagnosed with degenerative spine disease. Repeated magnetic resonance (MR) T1-weighted images of the thoracic spine suggested a subacute intramedullary hematoma. Due to progression of the neurological deficit, the patient was referred to a neurosurgeon, who indicated surgical evacuation of the hematoma. The intraoperative finding revealed an intramedullary spinal cord lipoma, which was later confirmed by histological analysis. Since subacute intramedullary hematomas and intramedullary spinal cord lipomas present with similar clinical and radiological features, diffusion-weighted MR imaging should be used to distinguish these entities.

Tentorial alignment and its relationship to cisternal dimensions of the pineal region: MRI anatomical study with surgical implications using the new clivotentorial method.

OBJECTIVES: Tentorial alignment and dimensions of posterior fossa cisterns are measurements whose variability can decrease surgical freedom if not taken into account when choosing the approach to the pineal region. The aim is to provide quantitative anatomical information regarding these dimensions, and to discuss their relevance in two most commonly used approaches to this region: the occipital transtentorial and supracerebellar-infratentorial approach. PATIENTS AND METHODS: A retrospective study of midsagittal T1-weighted MRI images of 410 randomly selected healthy subjects was performed. The clivus-tentorium (C-T) angle was measured to assess tentorial alignment. The following distances were used as craniocaudal cisternal measurements: quadrigeminal cistern = superior colliculi - inferior part of the splenium of corpus callosum (SC-ISCC), and superior cerebellar cistern = vermis - inferior part of the splenium of corpus callosum (VER-ISCC). RESULTS: Median C-T angle value was 19 ±â€¯7°, the quadrigeminal cistern height 6.7 ±â€¯1.6 cm, and the superior cerebellar cistern height 10.4 ±â€¯2.6 cm. The C-T angle was negatively correlated with the SC-ISCC distance (r = -0.271; p <  0.001) and the VER-ISCC distance (r = -0.052, p >  0.001). The SC-ISCC distance was positively correlated with the VER-ISCC distance (r = 0.282; p < 0.001). CONCLUSION: Our new method of measuring tentorial alignment provides a simple and effective aid in preoperative planning. For the first time, we present data on craniocaudal dimensions of posterior fossa cisterns, their relationship with tentorial alignment, and discuss their relevance in SCIT and OT approaches.

Expression patterns of Wnt signaling component, secreted frizzled­related protein 3 in astrocytoma and glioblastoma.

Secreted frizzled-related protein 3 (SFRP3) is a member of the family of soluble proteins, which modulate the Wnt signaling cascade. Novel research has identified aberrant expression of SFRPs in different types of cancer. In the present study the expression intensities and localizations of the SFRP3 protein across different histopathological grades of astrocytic brain tumors were investigated by immunohistochemistry, digital scanning and image analysis. The results demonstrated that the differences between expression levels and malignancy grades were statistically significant. Tumors were classified into four malignancy grades according to the World Health Organization guidelines. Moderate (P=0.014) and strong (P=0.028) nuclear expression levels were significantly different in pilocytic (grade I) and diffuse (grade II) astrocytomas demonstrating higher expression values, as compared with anaplastic astrocytoma (grade III) and glioblastoma (grade IV). When the sample was divided into two groups, the moderate and high cytoplasmic expression levels were observed to be significantly higher in glioblastomas than in the group comprising astrocytoma II and III. Furthermore, the results indicated that high grade tumors were associated with lower values of moderate (P=0.002) and strong (P=0.018) nuclear expression in comparison to low grade tumors. Analysis of cytoplasmic staining demonstrated that strong cytoplasmic expression was significantly higher in the astrocytoma III and IV group than in the astrocytoma I and II group (P=0.048). Furthermore, lower grade astrocytomas exhibited reduced membranous SFRP3 staining when compared with higher grade astrocytomas and this difference was statistically significant (P=0.036). The present results demonstrated that SFRP3 protein expression levels were decreased in the nucleus in higher grade astrocytoma (indicating the expected behavior of an antagonist of Wnt signaling), whereas when the SFRP3 was located in the cytoplasm an increased expression level of SFRP3 was identified in the high grade astrocytomas when compared with those of a low grade. This may suggest that SFRP3 acts as an agonist of Wnt signaling and promotes invasive behavior.