Antineoplastic multi-drug chemotherapy to sensitize tumors triggers multi-drug resistance and inhibits efficiency of maintenance treatment inglioblastoma cells.

Combinations of the well-known antineoplastic agents 5-fluorouracil (5-Fu), cisplatin, and paclitaxel are employed to increase radiotherapy/immunotherapy efficacy against persistent and resistant tumors. However, data remains needed on the hormetic, chronic, and long-term side effects of these aggressive combination chemotherapies. Here we investigated cellular and molecular responses associated with these combined agents, and their potential to induce multi-drug resistance against the temozolomide (TMZ) and etoposide (EP) used in glioblastoma maintenance treatment. We analyzed resistance and survival signals in U87 MG cells using molecular probes, fluorescent staining, qRT-PCR, and immunoblot. Repeated treatment with combined 5-Fu, cisplatin, and paclitaxel induced cross-resistance against TMZ and EP. Resistant cells exhibited elevated gene/protein expression of MRP1/ABCC1, ABCC2, BRCP/ABCG2, and GST. Moreover, they managed oxidative stress, cell cycle, apoptosis, and autophagy signaling to ensure survival. In these groups TMZ and etoposide efficiency dramatically reduced. Our result suggests that combined high-dose treatments of classical antineoplastic agents to sensitize tumors may trigger multi-drug resistance and inhibit maintenance treatment. When deciding on antineoplastic combination therapy for persistent/resistant glioblastoma, we recommend analyzing the long-term hormetic and chronic effects on cross-resistance and multi-drug resistance in primary cell cultures from patients. See also the Graphical Abstract(Fig. 1).

The Effect of Cerebral Oxygen Saturation Changes on Early Postoperative Neuropsychological Function in Patients Undergoing Cranial Surgery.

AIM: To compare the incidence of postoperative neuropsychological dysfunction in patients managed with cerebral saturation monitoring versus traditional approaches. MATERIAL AND METHODS: A hundred patients undergoing elective intracranial surgery were divided into two groups to receive intraoperative management via cerebral saturation monitoring (Group O) or the conventional approach (Group C). The postoperative neuropsychological function was evaluated by the antisaccadic eye movement test (ASEM) and the Mini-Mental State Examination (MMSE). These tests were performed preoperatively and postoperatively on the first, second, and fifth days. The time for the modified Aldrete score to reach 9 (MAS 9), adverse effects, and pain using a Visual Analog Scale (VAS) scores were recorded. RESULTS: Patient characteristics and surgery data were not statistically different. The MAS 9 of group O was significantly lower than that of group C (p < 0.001). The MMSE at the postoperative 1, 2, and 5 days were significantly higher in Group O compared to Group C (p < 0.001). ASEM was similar between groups. Group O was subdivided according to the type of surgery applied with diagnosis, and there were no statistically significant between-group differences in terms of areas under the curve for the cerebral regional oxygen saturation. There was no between-group difference regarding the mean arterial pressure at any time perioperatively. The heart rate at 80, 90, 100, and 110 min intraoperatively was significantly higher in group C than in Group O. CONCLUSION: Intraoperative cerebral oxygenation monitoring can reduce patient mortality and morbidity by allowing early postoperative neurological evaluation to detect potential neurocognitive deficits.

Effects of Expression of Matrix Metalloproteinases and Discoidin Domain Receptors in Ligamentum Flavum Fibrosis in Patients with Degenerative Lumbar Canal Stenosis.

STUDY DESIGN: This is a retrospective cohort study. PURPOSE: This study aimed to clarify the role of crosstalk between discoidin domain receptors (DDRs) and matrix metalloproteinases (MMPs) in the ligamentum flavum (LF) fibrosis obtained from patients with degenerative lumbar canal stenosis (DLCS). OVERVIEW OF LITERATURE: The DDRs, DDR1 and DDR2, are cell surface receptors and have an essential role in collagen fiber accumulation in several fibrotic diseases. MMPs are one of the critical factors in extracellular matrix remodeling and elastic fiber degradation in LF tissues. However, the crosstalk between DDRs and MMPs and the role of this molecular signal in LF fibrosis remain unclear. METHODS: A total of 35 patients were divided into two groups in this study. Spinal surgery was performed in 23 of these patients with the diagnosis of DLCS. Twelve patients with lumbar disk herniation (LDH) were included in the control group. On axial T2-weighted magnetic resonance imaging, LF thickness was measured bilaterally at the level of the facet joint. Histology, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot analyses were performed on LF tissue samples. LF tissues were stained with hematoxylin and eosin. In addition, the grade of fibrosis was histologically assessed using Masson trichrome triple staining. DDR1 and DDR2 Western blot analyses were performed. DDR1, DDR2, MMP2, MMP3, MMP9, and MMP13 expression levels were measured using qRT-PCR analysis. RESULTS: The grade of fibrosis and LF thickness were significantly higher in the DLCS patients than in the LDH patients. DDR1 and DDR2 gene expression and protein levels in LF tissues are significantly greater in DLCS samples than in control samples, according to both qRT-PCR and Western blot analyses. In addition, we detected a significant expression of the MMP3, MMP9, and MMP13, which are known to have important roles in extracellular matrix remodeling in DLCS. Furthermore, we discovered a link between DDR protein levels and LF thickness, fibrosis, and MMP3/MMP9. CONCLUSIONS: Our results indicate that DDR1, DDR2, and MMP3 and MMP9 signals can be correlated with each other in LF tissues and be promoted LF fibrosis leading to spinal canal narrowing in patients with DLCS.

The role of melatonin in angio-miR-associated inhibition of tumorigenesis and invasion in human glioblastoma tumour spheroids.

Micro-RNA (miRNA)-based regulation of hypoxia, angiogenesis and tumour growth provides promising targets for effective therapy in malignant glioblastoma multiforme (GBM). Accumulating evidence suggests a potential role of melatonin in miRNA expression in cancer cells. Despite these findings, the melatonin-miRNA interaction in GBM and the effect of this interaction on GBM tumour development and invasion are not clearly understood. The aim of the present study was to evaluate the effects of melatonin on human GBM tumour spheroid tumorigenesis and invasion in vitro, and to analyse the interaction between 36 angio-miRNAs and the HIF1/VEGF/MMP9 axis, which is known to be associated with the antitumour effect of melatonin. We found that melatonin is able to selectively induce cell death in single-layer U87-MG cells (a GBM cell line) in a dose- and time-dependent manner, as characterized by MTT assay. The use of tumour spheroids and a Matrigel invasion assay revealed that melatonin impairs tumorigenesis, and it significantly reduced both the tumour spheroid area and invasion rate, especially at the 0.5 mM and 1 mM concentrations. This inhibition was accompanied by strong reductions in hypoxia-inducible factor 1-α (HIF1-α) and vascular endothelial growth factor (VEGF) gene expression and protein levels in GBM tumour spheroids. In addition, melatonin significantly reduced the relative gene expression and protein levels of matrix metalloproteinase-9 (MMP-9). This study revealed that six differentially expressed angio-miRs (miR-15b, miR-18a-5p, miR-23a-3p, miR-92a-3p, miR-130a-5p, miR-200b-3p) may play important roles in GBM tumorigenesis and invasion, and all respond to melatonin therapy. Our results suggest that melatonin inhibits tumorigenesis and invasion of human GBM tumour spheroids, possibly by suppressing HIF1-α/VEGF/MMP9 signalling via regulation of angio-miRNAs.

The Dose Dependent Effects of Ruxolitinib on the Invasion and Tumorigenesis in Gliomas Cells via Inhibition of Interferon Gamma-Depended JAK/STAT Signaling Pathway.

OBJECTIVE: Glioblastoma multiforme (GBM) is the most aggressive for of brain tumor and treatment often fails due to the invasion of tumor cells into neighboring healthy brain tissues. Activation of the Janus kinase-signal transducer and activator of transcription (JAK/STAT) signaling pathway is essential for normal cellular function including angiogenesis, and has been proposed to have a pivotal role in glioma invasion. This study aimed to determine the dose-dependent effects of ruxolitinib, an inhibitor of JAK, on the interferon (IFN)-I/IFN-α/IFN-ß receptor/STAT and IFN-γ/IFN-γ receptor/STAT1 axes of the IFN-receptor-dependent JAK/STAT signaling pathway in glioblastoma invasion and tumorigenesis in U87 glioblastoma tumor spheroids. METHODS: We administered three different doses of ruxolitinib (50, 100, and 200 nM) to human U87 glioblastoma spheroids and analyzed the gene expression profiles of IFNs receptors from the JAK/STAT pathway. To evaluate activation of this pathway, we quantified the phosphorylation of JAK and STAT proteins using Western blotting. RESULTS: Quantitative real-time polymerase chain reaction analysis demonstrated that ruxolitinib led to upregulated of the IFN-α and IFN-γ while no change on the hypoxia-inducible factor-1α and vascular endothelial growth factor expression levels. Additionally, we showed that ruxolitinib inhibited phosphorylation of JAK/STAT proteins. The inhibition of IFNs dependent JAK/STAT signaling by ruxolitinib leads to decreases of the U87 cells invasiveness and tumorigenesis. We demonstrate that ruxolitinib may inhibit glioma invasion and tumorigenesis through inhibition of the IFN-induced JAK/STAT signaling pathway. CONCLUSION: Collectively, our results revealed that ruxolitinib may have therapeutic potential in glioblastomas, possibly by JAK/STAT signaling triggered by IFN-α and IFN-γ.

The Role of JAK-STAT Signaling Activation in Hypertrophied Ligamentum Flavum.

BACKGROUND: Although previous studies have reported the expression of JAK1, STAT3, and phosphorylated STAT3 in hypertrophied ligamentum flavum (LF), the role of the Janus kinase-signal transducer and activator of transcription (JAK/STAT) signaling pathway in hypertrophied LF has not been fully elucidated. The aim of this study was to identify the important JAK/STAT gene expression patterns of the 3 main receptors involved in this pathway: interferon (IFN)-γ receptor (IFN-γR), IFN-α receptor (IFNAR), and interleukin (IL)-6 receptor (IL-6R). METHODS: The human LF specimens were obtained from 28 patients who underwent lumbar spine surgery for either degenerative lumbar canal stenosis (DLCS) (n = 28) or lumbar disc herniation (LDH) (n = 20). In this design, patients with LDH served as the control group. The degree of fibrosis was demonstrated by Masson's trichrome staining. The location and expression profiling of the JAK/STAT pathway were analyzed by quantitative real-time polymerase chain reaction and Western blotting. The thickness of the LF was measured with axial T1-weighted magnetic resonance imaging. RESULTS: The most severe fibrotic changes were on the dorsal side of the LF. IL-6 and IFN-I expression levels were significantly increased on the dorsal side of the LF. While expression levels of IL-6R and IFNAR on the dural and dorsal side were significantly higher in the DLCS samples, IFN-γR and endothelial epidermal growth factor receptor in LF samples showed a significant increase only on the dorsal side. JAK/STAT genes were significantly expressed, especially on the dorsal side. CONCLUSIONS: Our data suggest that IFNAR- and IL-6R-dependent JAK/STAT signaling pathways may be significant targets in drug development strategies for the treatment of LF hypertrophy.

Inhibition of the Invasion of Human Glioblastoma U87 Cell Line by Ruxolitinib: A Molecular Player of miR-17 and miR-20a Regulating JAK/STAT Pathway.

AIM: To determine the interaction between ruxolitinib, JAK/STAT signalling, and two angio-microRNAs (miRs) to expose potential target molecules in the inhibition of glioblastoma invasion. MATERIAL AND METHODS: The invasion properties of glioblastoma were analyzed using a cancer cell spheroid invasion assay. Following treatment of 195 nM ruxolitinib, the relative expression levels of miR-17 and miR-20a and genes of IL-6/JAK/STAT3 receptor signaling belonging to the JAK/STAT pathway were measured by qRT-PCR in treated and untreated three-dimensional tumor spheres of U87 cells. RESULTS: Our results indicated that a therapeutic dose of ruxolitinib (195 nM) significantly increased miR-17 and miR-20a expression. Ruxolitinib treatment resulted in the production of IL-6 and active formation of IL-6 receptor complex for the subsequent activation of the IL-6R/JAK2/STAT3 axis. However, ruxolitinib treatment significantly decreased the expression of JAK2 and PI3K. Pearson correlation analyses revealed a strong negative correlation of miR-17 with JAK2, STAT3, and PI3K expressions, and also miR-20a has a negative correlation with expression levels of JAK2 and PI3K. The only positive correlation was found to be between miR-20a and IL-6, gp130 expressions. CONCLUSION: The specific JAK2 inhibitor ruxolitinib plays an important role in glioblastoma angiogenesis biology via inhibiting IL-6 receptor-dependent JAK/STAT signaling. Additionally, both miR-17a-3p and miR-20a overexpression induced by ruxolitinib treatment may be playing a major role in downregulated JAK2, STAT3, and PI3K proteins. Our results suggest that miR-17-3p and miR-20a-5p may serve as new therapeutic targets for the treatment of glioblastoma.

Spontaneous subarachnoid haemorrhage incidence among hospitalised patients in Edirne, Turkey.

BACKGROUND: To the best of our knowledge, no data has been published about the spontaneous subarachnoid haemorrhage (sSAH) incidence in Turkey. We aimed to report the estimation of sSAH incidence in Edirne Province, in Turkey for the first time, using the data acquired from a single medical centre which has the biggest and the most comprehensive emergency department in Edirne and to where a great majority of patients are referred. METHODS: We investigated all the accessible sSAH patients' data obtained from computer-based automation systems and all the written documents in the neurosurgery and the emergency departments. Patients included in the study were diagnosed with sSAH between the dates of January 2007 and December 2011 and were resident in Edirne. We used this data to calculate the crude and age-adjusted incidence rates of sSAH for every decade. RESULTS: One hundred fifty-four patients have been diagnosed with sSAH during a 5-year period. Among them, 72 were men (47.8%) and 82 (53.2%) were women. The mean age of the patients was 60.8 years and age range was 23-85 years. The overall annual adjusted incidence rate for sSAH was 10.3 per 100,000 person-years (95% confidence interval = 10.2-10.3). Annual adjusted incidence rate was 10 per 100,000 person-years (95% confidence interval = 10-10.1) for men. For women, it was 10.4 per 100,000 person-years (95% confidence interval = 10.4-10.5). For both sexes, after the 6th decade, the annual incidence rate of sSAH was higher than 10 per 100,000, reaching over 20 per 100,000 person-years after the 7th decade. The overall crude incidence rate for sSAH was 10.3 per 100,000 person-years. For men, the crude incidence rate was 9.4 per 100,000 person-years and for women, it was 11.2 per 100,000 person-years. CONCLUSIONS: This study showing the first sSAH incidence estimation in Edirne might also be accepted as an estimation of overall epidemiological sSAH aspect in Turkey. Future investigations should be realised in different parts of Turkey to enlighten the epidemiological state of affairs and the course of sSAH in Turkey.

Incomplete Isolated C7 Root Injury Caused by Gunshot Wound: A Case Report.

Gunshot wounds to the spine cause severe neurological and/or internal organs damages. Although most of publications in the literature are realized on military injuries, increased civilian arming which raises civilian gunshot injuries is a new social danger causing serious health problems. In gunshot injuries to the spine; vertebral column, spinal cord and nerve roots are damaged with direct, indirect and transient cavitation related mechanisms. In this case report, we present 24 years old male patient who had severe pain and monoparesis in left upper extremity followed by gunshot injury to the spine with clinical, radiological and postoperative follow-up findings.

Efficiacy of resveratrol and quercetin after experimental spinal cord injury.

BACKGROUND: The aim of this study was to investigate the effect of natural antioxidants resveratrol and quercetin on oxidative stress and secondary cell damage in rats with acute spinal cord injury. METHODS: In this experimental study, 42 male Sprague-Dawley rats were used. Spinal cord injury was performed with clip compression method at level of T4-5. The study was conducted using 6 groups: control, trauma, trauma and solvent, trauma and resveratrol, trauma and quercetin, and trauma with combined resveratrol and quercetin. All rats were euthanized 48 hours after the procedure. Effects of resveratrol and quercetin on serum and tissue total antioxidant capacity and paraoxanase activity level were examined. RESULTS: Compared to trauma group, there was a significant increase in total antioxidant capacity and paraoxanase activity level in resveratrol, quercetin, and combined treatment groups. There was no significant difference between resveratrol and quercetin groups with regard to total antioxidant capacity and paraoxanase activity level. Total antioxidant capacity and paraoxanase activity level were significantly higher in solvent group than trauma group. In histopathological evaluation, there was a decrease in polymorphonuclear leukocyte infiltration in solvent, resveratrol, quercetin, and combined treatment groups. CONCLUSION: Biochemical and histological staining results of present study showed that resveratrol and quercetin may be effective in preventing secondary damage in spinal cord injury.

Perimesencephalic subarachnoid hemorrhage: Etiologies, risk factors, and necessity of the second angiogram.

AIM: In this paper, we aim to present our experience with a series of patients with PMSAH. In addition, the clinical course of perimesencephalic subarachnoid hemorrgade (PMSAH) is discussed with an evaluation of etiologies, risk factors, and the necessity for a second angiogram on follow-up. MATERIALS AND METHODS: The data for this study were obtained retrospectively from patients who were treated at the Uludag University, School of Medicine, Department of Neurosurgery, Division of Neurovascular Surgery's clinic with a diagnosis of PMSAH between January 1980 and March 2002. RESULTS: We identified a total of 24 patients, 12 male. The mean age at the time of hemorrhage was 53 ± 12 years. In all patients, the onset was typical with a sudden severe headache. Five of the patients were Hunt-Hess Grade I, 15 were Grade II, and 4 were Grade III. The initial 4-vessel angiography was normal in 23 cases. Twenty-two had a second 4-vessel angiography, and all were normal. We observed acute hydrocephalus in 5 patients (20.8%). We did not observe re-bleeding during the follow-up of our patients. CONCLUSION: Patients with PMSAH have a particularly excellent outcome, and there is no need to evaluate these patients with repeat angiography.

True aneurysm of superficial temporal artery accompanying multiple intracranial aneurysm.

Superficial temporal artery (STA) aneurysms are very infrequent. Moreover, true aneurysms, which are not pseudoaneurysms associated with trauma or previous surgery are even rarer. With this manuscript, authors present a case of a 79-year-old woman suffering from subarachnoid hemorrhage whose radiological examinations revealed multiple intracranial aneurysms along with an STA aneurysm. This very rare case, to the best of our knowledge, the second case reported so far, might contribute to the literature and lead further investigations toward the rare association between intracranial aneurysms and STA aneurysms.

Therapeutic efficacy of tadalafil and eriythropoietin in experimental spinal cord injury.

BACKGROUND: This experimental study was an investigation of the efficacy of erythropoietin and tadalafil in rats with induced spinal cord injury (SCI). METHODS: Thirty-five Sprague Dawley rats were distributed into 5 groups. First group was used for normal biochemical values. Spinal cord injury was induced in 4 remaining groups with clip compression technique after laminectomy process to T10 vertebra. Second group was designated solvent group and received 1 cc physiological serum after injury. Third group was medicated with intraperitoneal 2000 u/kg single dose erythropoietin after injury. Orogastric 2 mg/kg single dose tadalafil was administered to fourth group after injury. Fifth group did not receive any treatment and was used for biochemical values with injury. All subjects were sacrificed 48 hours after application. Malondialdehyde (MDA) and total antioxidant capacity (TAOC) values were evaluated using blood and tissue samples. RESULTS: Lowest serum and tissue MDA values were found in group with erythropoietin intake. While highest serum TAOC values of all groups were seen in tadalafil group, highest tissue TAOC values were observed in group given erythropoietin. CONCLUSION: It was concluded that by decreasing oxidative stress, tadalafil and erythropoietin can inhibit secondary damage in SCI.

Symptomatic foraminal extradural meningeal cyst.

The authors described the case of a 39-year-old man with Klippel-Trénaunay syndrome, who had an extradural meningeal cyst expanding into intervertebral foramen of lumbar 2 and 3 vertebrae. The patient suffered from low back pain radiating to the left lower extremity. Magnetic resonance imaging revealed a huge extradural meningeal cyst growing through intervertebral foramen far laterally. A widened neural foramen of L2 and L3 vertebrae was observed on his plain radiography. The cyst was totally excised after tying the ostium connecting the subarachnoid space of the dural sac. This case supports the congenital theory in the pathogenesis of spinal cysts because the Klippel-Trénaunay syndrome is a congenital disorder including a mesodermal abnormality which may be the causative factor for a congenital defect in dura.