Altered Hemodynamics Associated with Pathogenesis of the Vertebral Artery Dissecting Aneurysms

The etiology of the vertebral dissecting aneurysms is largely unknown, and they frequently occurs in relatively healthy young men. Objectives and Methods. A series of 57 consecutive cases defined by angiography were evaluated with regard to deviation in the course of the affected and contralateral vertebral arteries. Division was into 3 types: Type I without any deviation, Type II with mild-to-moderate deviation but not over the midline; and Type III with marked deviation over to the contralateral side beyond the midline. Results. The most frequent type of VA running was Type III for the affected and Type I nonaffected side, with this being found in all 17 patients except one. All of the Type III dissections occurred just proximal to a tortuous portion, while in cases with Type-I- and Type-II-affected sides, the majority (33 of 39) occurred near the union of the vertebral artery. In 10 of 57, a non-dominant side was affected, all except one being of Type I or II. With 12 recent patients assessed angiographically in detail for hemodynamics, eleven patients showed contrast material retrograde inflowing into the pseudolumen from the distal portion of the dissection site. Turbulent blood flow was recognized in all of these patients with retrograde inflow. Conclusions. Turbulent blood flow is one etiology of vertebral artery dissection aneurysms, with the sites in the majority of the cases being just proximal to a tortuous portion or union of vessels. In cases with dissection proximal to the tortuous course of the vertebral artery, retrograde inflow will occur more frequently than antegrade, which should be taken into account in designing therapeutic strategies

An Operation in the Park Bench Position Complicated by Massive Tongue Swelling

This paper presents a case of massive tongue swelling as a complication after an operation in the park bench position. A 43-year-old male who had undergone a resection of a mass in the petrous bone of the clivus showed massive tongue swelling after the surgery in the left park bench position. A direct compression of the bite block caused the swelling of tongue. Tongue swelling may become fatal if it progresses to an airway obstruction; therefore the intraoperative and postoperative management is important

ALK signaling cascade confers multiple advantages to glioblastoma cells through neovascularization and cell proliferation

<div><p>Anaplastic lymphoma kinase (ALK), which is a receptor tyrosine kinase, is essentially and transiently expressed in the developing nervous system. Here we examined the functional role of the <i>ALK</i> gene in glioblastomas (GBMs). In clinical samples of GBMs, high ALK expression without gene rearrangements or mutations was frequently observed in perivascular lesions, in contrast to the relatively low expression in the perinecrotic areas, which was positively correlated with N-myc and phosphorylated (p) Stat3 scores and Ki-67 labeling indices. ALK immunoreactivity was also found to be associated with neovascular features including vascular co-option and vascular mimicry. In astrocytoma cell lines, cells stably overexpressing full-length ALK showed an increase in expression of pStat3 and pAkt proteins, as well as hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor-A (VEGF-A) mRNAs, in contrast to cells with knockdown of endogenous ALK which showed decreased expression of these molecules. Transfection of the constitutively active form of Stat3 induced an increase in <i>HIF-1</i>α promoter activity, and the overexpression of HIF-1α in turn resulted in enhancement of <i>VEGF-A</i> promoter activity. In addition, cells with overexpression or knockdown of ALK also showed a tendency toward increased and decreased proliferation, respectively, through changes in expression of pAkt and pStat3. Finally, <i>ALK</i> promoter was significantly activated by transfection of Sox4 and N-myc, which are known to contribute to neuronal properties. These findings therefore suggest that N-myc/Sox4-mediated ALK signaling cascades containing Stat3, Akt, HIF-1α, and VEGF-A confer multiple advantages to tumor growth through alterations in neovascularization and cell proliferation in GBMs.</p></div

Correlations among IHC markers investigated in GBM cases.

<p>Correlations among IHC markers investigated in GBM cases.</p

Relationship between ALK expression and neovascularization in GBMs.

<p>Relationship between ALK expression and neovascularization in GBMs.</p

Up-regulation of ALK expression in hypervascular areas of GBMs.

<p>(A) Left: staining by hematoxylin and eosin (HE) and IHC for ALK in GBMs. Note the strong cytoplasmic ALK (5A4) positivity in cells around vascular components (a) (indicated by arrows) of GBMs, in contrast to the sporadic staining or absence in tumor cells adjacent to necrotic foci (b) (partitioned by dotted line). Panels (a) and (b) are magnified views of the boxed areas. Original magnification, x100 (left) and x200 (middle and right). Right: IHC score for ALK (5A4) in perivascular (Peri-v) and perinecrotic (Peri-n) areas. (B) Staining by HE (upper and lower left) and double-staining for ALK/CD34 (upper and lower middle) and SMA/CD34 (upper and lower right) in mature vessels (upper) and microvessels (lower) in GBM. Strong ALK (5A4) (brown) positivity is observed in GBM cells surrounding CD34 (red)-positive mature vessels (upper middle), while focal ALK (5A4) immunoreactivity is also evident in CD34-positive microvessels (lower middle). Note the close association between SMA- and CD34-positive cells in both tumor vasculatures (upper and lower left). Insets show magnified views of the boxed areas. Original magnification, x200 and x400 (inset). (C) Left: staining by HE and IHC for ALK and CD34 in GBMs. Note the strong ALK (5A4) immunopositivity in areas with high CD34 immunoreactivity, in contrast to the sporadic staining or absence in the low immunoreactivity lesions. Original magnification, x100. Right: relationship between ALK (5A4) immunointensity and microvascular density as determined by CD34 immunoreactivity in GBMs. The data shown are means±SDs. (D) Western blot analysis of the indicated proteins after CoCl₂ treatment with the different doses shown for 24 hours (left) and 50 μM CoCl₂ for the time shown (right) in KINGS-1 cells.</p

Association between ALK expression and cell proliferation in astrocytoma cells.

<p>(A) Left: KS-ALK#4 cells and mock cells were seeded at low density. The cell numbers are presented as means±SDs. P0, P2, P4, P6, and P8 indicate 0, 2, 4, 6, and 8 days after cell passage, respectively. Right: Cell Counting Kit-8 (CCK-8) assay for cell proliferation. Cells were seeded at 1x10³ cells in 96-well plates. Viable cell numbers were quantitated. Relative absorbance values (P5 or P7 relative to P2) are presented as means±SDs. P2, P5, and P7 indicate 2, 5, and 7 days after cell passage, respectively. This experiment was performed in triplicate using independent samples. (B) Western blot analysis for the indicated proteins in ALK#4 and mock cells after serum stimulation for the times shown. (C) Left: KINGS-shALK#37, #46 cells, and mock cells were seeded at low density. The cell numbers are presented as means±SDs. P0, P3, P6, and P9 indicate 0, 3, 6, and 9 days after cell passage, respectively. Right: CCK-8 assay for cell proliferation to quantitate viable cell numbers as mentioned above. (E) Staining by hematoxylin and eosin (HE) and IHC for pStat3, pAkt, and Ki-67 in GBMs. Note the strong immunoreactivity for these molecules in perivascular lesions (vessels are indicated by arrows), in contrast to the weak immunoreaction in perinecrotic areas (necrotic lesion is partitioned by dotted line). Original magnification, x100. (F) IHC scores for pStat3 and pAkt and Ki-67 labeling indices in perivascular (Pv) and perinecrotic (Pn) lesions.</p

Relationship of ALK expression with Sox4 and N-myc in astrocytomas.

<p>(A) Various <i>ALK</i> promoter constructs used in this study. (B) KS-1 cells were transfected with <i>ALK</i> promoter constructs, together with the indicated <i>Sox</i> genes. Relative activity was determined based on arbitrary light units of luciferase activity normalized to pRL-TK activity. The activities of the reporter plus the effector relative to that of the reporter plus empty vector are shown as means±SDs. The experiment was performed in duplicate. (C) KS-1 cells were transfected with <i>ALK</i> promoter constructs, together with either Sox4, c-myc, or N-myc. (D) Various promoter constructs were used for evaluating transcriptional regulation of the <i>ALK</i> promoter by either Sox4 or N-myc. (E) The shortest <i>ALK</i> promoter constructs containing mutations in two putative E-boxes (E1 and E2), along with either Sox4 or N-myc, were transfected into KS-1 cells. (F) The Sox4 (left) and the N-myc (right) promoter constructs, along with either c-myc, N-myc, or Sox4, were transfected into KS-1 cells. (G) Left: staining by hematoxylin and eosin (HE) and IHC for N-myc and c-myc in GBMs. Note the strong immunoreactivity for N-myc, but not c-myc, in tumor cells around vessel (indicated by arrows) but not perinecrotic area (necrotic lesion is partitioned by dotted line). Right: IHC scores for N-myc and c-myc in perivascular (Peri-v) and perinecrotic (Peri-n) lesions.</p

ALK/Stat3/HIF-1α axis in astrocytoma cells.

<p>Western blot (left) and RT-PCR (right) analyses for the indicated molecules in (A) KS-ALK#4 cells and (B) KINGS-shALK#37 and #46 cells. (C) KS-1 cells were transfected with HIF-1α (left) and VEGF-A (right) reporter constructs, together with either ALK, Stat3C, or HIF-1α. Relative activity was determined based on arbitrary light units of luciferase activity normalized to pRL-TK activity. The activities of the reporter plus the effector relative to that of the reporter plus empty vector are shown as means±SDs. The experiment was performed in duplicate. v, empty vector. Western blot (left) and RT-PCR (right) analyses for the indicated molecules in (D) KS-ALK#4 cells and (E) KINGS-shALK#37 and #46 cells after CoCl₂ treatment for 4 hours. (F) Left: staining by hematoxylin and eosin (HE) and IHC for HIF-1α in GBMs. Note the strong HIF-1α immunoreactivity (indicated by arrows) in both perivascular areas and pseudopalisading around necrotic lesion (partitioned by dotted line). Original magnification, x100. Right: IHC score for HIF-1α in perivascular (Peri-v) and perinecrotic (Peri-n) areas of GBMs.</p

Association between ALK expression and neovascular features in GBMs.

<p>(A) Staining by hematoxylin and eosin (HE) and IHC for ALK in vascular co-option. Note the strong ALK (5A4) immunoreactivity in vascular co-option with classic perivascular cuffs (indicated by arrows). Insets show magnified view of the boxed area. Original magnification, x200 and x400 (inset). (B) Staining by HE and IHC for ALK and CD34 in vascular mimicry. Note the diffuse ALK (5A4) immunoreaction and focal CD34 immunoreactivity in the perfused vascular networks containing red blood cells. The lower right panel shows the magnified view of the boxed area in the lower left panel. Original magnification, x100 and x400 (lower right panel). (C) Upper: staining by HE and IHC for ALK. Note the strong ALK (5A4) immunoreactivity in tumor vasculature (indicated by boxes and magnified in the inset), as well as tumor cells around vascular components. Original magnification, x200 and x400 (inset). Middle and lower: staining by HE and ISH for ALK mRNA in GBMs. Note the positive ALK mRNA signals in tumor vasculature which are indicated by boxes in middle right panel and magnified in lower right panels (positive ALK mRNA signal in tumor vasculature are indicated by arrows), as well as tumor cells around vascular components. Original magnification, x200 and x400 (lower left panel).</p