Analysis of Tumor Angiogenesis and Immune Microenvironment in Non-Functional Pituitary Endocrine Tumors

Cavernous sinus (CS) invasion is an aggressive behavior exhibited by pituitary neuroendocrine tumors (PitNETs). The cause of CS invasion in PitNETs has not been fully elucidated. The tumor immune microenvironment, known to promote aggressive behavior in various types of tumors, has not been examined for PitNETs. Vascular endothelial growth factor (VEGF)/VEGF receptor (VEGFR) signaling is strongly associated with the tumor immune microenvironment. In the present study, these molecular and histopathological characteristics were examined in invasive non-functional PitNETs (NF-PitNETs). Twenty-seven patients with newly diagnosed NF-PitNETs (with CS invasion: 17, without CS invasion: 10) were analyzed by immunohistochemistry for VEGF-A/VEGFR1 and 2, hypoxia-inducible Factor (HIF), tumor-infiltrating lymphocytes, immunosuppressive cells including regulatory T cells (Tregs) and tumor-associated macrophages (TAMs), and immune checkpoint molecules. Previously validated tumor proliferation markers including mitotic count, Ki-67 index, and p53 were also analyzed for their expressions in NF-PitNETs. VEGF-A and VEGFR1 were expressed on not only vascular endothelial cells, but also on tumor cells. The expressions of VEGF-A and VEGFR1 were significantly higher in NF-PitNETs with CS invasion. The number of TAMs and the expression of PD-L1 were also significantly higher in NF-PitNETs with CS invasion than in NF-PitNETs without CS invasion. The high expression of VEGF-A and VEGFR1 and associated immunosuppressive microenvironment were observed in NF-PitNETs with CS invasion, suggesting that a novel targeted therapy can be applied

Effect of Audiovisual Cross-Modal Conflict during Working Memory Tasks : A Near-Infrared Spectroscopy Study

Cognitive conflict effects are well characterized within unimodality. However, little is known about cross-modal conflicts and their neural bases. This study characterizes the two types of visual and auditory cross-modal conflicts through working memory tasks and brain activities. The participants consisted of 31 healthy, right-handed, young male adults. The Paced Auditory Serial Addition Test (PASAT) and the Paced Visual Serial Addition Test (PVSAT) were performed under distractor and no distractor conditions. Distractor conditions comprised two conditions in which either the PASAT or PVSAT was the target task, and the other was used as a distractor stimulus. Additionally, oxygenated hemoglobin (Oxy-Hb) concentration changes in the frontoparietal regions were measured during tasks. The results showed significantly lower PASAT performance under distractor conditions than under no distractor conditions, but not in the PVSAT. Oxy-Hb changes in the bilateral ventrolateral prefrontal cortex (VLPFC) and inferior parietal cortex (IPC) significantly increased in the PASAT with distractor compared with no distractor conditions, but not in the PVSAT. Furthermore, there were significant positive correlations between Delta task performance accuracy and Delta Oxy-Hb in the bilateral IPC only in the PASAT. Visual cross-modal conflict significantly impairs auditory task performance, and bilateral VLPFC and IPC are key regions in inhibiting visual crossmodal distractors

Transcranial direct current stimulation to the left dorsolateral prefrontal cortex enhances early dexterity skills with the left non-dominant hand : a randomized controlled trial

Background The left dorsolateral prefrontal cortex (DLPFC) is involved in early-phase manual dexterity skill acquisition when cognitive control processes, such as integration and complexity demands, are required. However, the effectiveness of left DLPFC transcranial direct current stimulation (tDCS) on early-phase motor learning and whether its effectiveness depends on the cognitive demand of the target task are unclear. This study aimed to investigate whether tDCS over the left DLPFC improves non-dominant hand dexterity performance and determine if its efficacy depends on the cognitive demand of the target task.Methods In this randomized, double-blind, sham-controlled trial, 70 healthy, right-handed, young adult participants were recruited. They were randomly allocated to the active tDCS (2 mA for 20 min) or sham groups and repeatedly performed the Purdue Pegboard Test (PPT) left-handed peg task and left-handed assembly task three times: pre-tDCS, during tDCS, and post tDCS.Results The final sample comprised 66 healthy young adults (mean age, 22.73 +/- 1.57 years). There were significant interactions between group and time in both PPT tasks, indicating significantly higher performance of those in the active tDCS group than those in the sham group post tDCS (p < 0.001). Moreover, a greater benefit was observed in the left-handed assembly task performance than in the peg task performance (p < 0.001). No significant correlation between baseline performance and benefits from tDCS was observed in either task.Conclusions These results demonstrated that prefrontal tDCS significantly improved early-phase manual dexterity skill acquisition, and its benefits were greater for the task with high cognitive demands. These findings contribute to a deeper understanding of the underlying neurophysiological mechanisms of the left DLPFC in the modulation of early-phase dexterity skill acquisition

Histochemical examination on principal collagen fibers in periodontal ligaments of ascorbic acid-deficient ODS-od/od rats

In this study, we aimed to clarify the role of ascorbic acid in collagen synthesis in periodontal ligaments using osteogenic disorder Shionogi (ODS)/ShiJcl-od/od rats lacking L-gulonolactone oxidase. These rats cannot synthesize ascorbic acid in vivo. Eight-week-old ODS/ShiJcl-od/od male rats were administered ascorbic acid solution at a concentration of 200 mg/dL (control group, n = 6) or ascorbic acid solution at concentration of 0.3 mg/dL (insufficient group, n = 12). Six rats of the insufficient group were then given with ascorbic acid solution at concentration of 200 mg/dL for additional 3 weeks (rescued group, n = 6), and then, their mandibles were histochemically examined. Consequently, the insufficient group specimens were seen to possess fewer collagen fibers, and silver impregnation revealed numerous fine, reticular fiber-like fibrils branching off from collagen in the periodontal ligaments. In control group, faint immunoreactivities for matrix metalloproteinase (MMP)2 and cathepsin H were seen in the periphery of blood vessels and throughout the ligament, respectively. In contrast, in the insufficient group, intense MMP2-immunoreactivity was observed to be associated with collagen fibrils in the periodontal ligaments, and cathepsin H-immunopositivity was seen in ligamentous cells. The rescued group showed abundant collagen fibers filling the periodontal ligament space. Under transmission electron microscopy, ligamentous fibroblasts incorporated collagen fibrils into tubular endosomes/lysosomes while simultaneously synthesizing collagen fibril bundles. Thus, ascorbic acid insufficiency affected the immunolocalization of cathepsin H and MMP2; however, ligamentous fibroblasts appear to possess the potential to synthesize collagen fibers when supplied with ascorbic acid

A Pilot Study of Vaccine Therapy with Multiple Glioma Oncoantigen/Glioma Angiogenesis-Associated Antigen Peptides for Patients with Recurrent/Progressive High-Grade Glioma

High-grade gliomas (HGGs) carry a dismal prognosis despite current treatments. We previously confirmed the safety and immunogenicity of a vaccine treatment targeting tumor angiogenesis with synthetic peptides, for vascular endothelial growth factor receptor (VEGFR) epitopes in recurrent HGG patients. In this study, we evaluated a novel vaccine therapy targeting not only tumor vasculature but also tumor cells, using multiple glioma oncoantigen (GOA)/glioma angiogenesis-associated antigen (GAAA) peptides in HLA-A2402+ recurrent/progressive HGG patients. The vaccine included peptide epitopes from four GOAs (LY6K, DEPDC1, KIF20A, and FOXM1) and two GAAAs (VEGFR1 and VEGFR2). Ten patients received subcutaneous vaccinations. The primary endpoint was the safety of the treatment. T-lymphocyte responses against GOA/GAAA epitopes and treatment response were evaluated secondarily. The treatment was well tolerated without any severe systemic adverse events. The vaccinations induced immunoreactivity to at least three vaccine-targeted GOA/GAAA in all six evaluable patients. The median overall survival time in all patients was 9.2 months. Five achieved progression-free status lasting at least six months. Two recurrent glioblastoma patients demonstrated stable disease. One patient with anaplastic oligoastrocytoma achieved complete response nine months after the vaccination. Taken together, this regimen was well tolerated and induced robust GOA/GAAA-specific T-lymphocyte responses in recurrent/progressive HGG patients

Analysis of the Anticipatory Behavior Formation Mechanism Induced by Methamphetamine Using a Single Hair

While the suprachiasmatic nucleus (SCN) coordinates many daily rhythms, some circadian patterns of expression are controlled by SCN-independent systems. These include responses to daily methamphetamine (MAP) injections. Scheduled daily injections of MAP resulted in anticipatory activity, with an increase in locomotor activity immediately prior to the time of injection. The MAP-induced anticipatory behavior is associated with the induction and a phase advance in the expression rhythm of the clock gene Period1 (Per1). However, this unique formation mechanism of MAP-induced anticipatory behavior is not well understood. We recently developed a micro-photomultiplier tube (micro-PMT) system to detect a small amount of Per1 expression. In the present study, we used this system to measure the formation kinetics of MAP-induced anticipatory activity in a single whisker hair to reveal the underlying mechanism. Our results suggest that whisker hairs respond to daily MAP administration, and that Per1 expression is affected. We also found that elevated Per1 expression in a single whisker hair is associated with the occurrence of anticipatory behavior rhythm. The present results suggest that elevated Per1 expression in hairs might be a marker of anticipatory behavior formation

The diversity of preosteoblastic morphology : Preosteoblastic response to parathyroid hormone

The current concept of a preosteoblast is a precursor of an osteoblast, which is regarded as a transient cell type during osteoblastic differentiation. We have previously demonstrated different phenotypes of preosteoblasts expressing Runx2, ALPase, and BrdU incorporation. Transmission electron microscopy revealed following four distinct preosteoblastic cell types : 1) cells rich in rough endoplasmic reticulum (rER) but with a few vesicles and vacuoles (ERrich/vesicle-poor preosteoblasts), 2) cells extending their cytoplasmic processes connecting distant cells, with a small amount of scattered cisterns of rER and many vesicles and vacuoles (ER-poor/vesicle-rich preosteoblasts), 3) translucent cells showing few dispersed cell organelles and irregular cell shape with a translucent cytoplasm (translucent cells), and 4) small cells without developed cell organelles (small undifferentiated cells). ER-rich/vesicle-poor preosteoblasts were often closely adjacent to mature osteoblasts and therefore appeared to be ready for differentiation into osteoblasts. In contrast, after the administration of parathyroid hormone (PTH), ER-poor/vesicle-rich preosteoblasts rather than ER-rich/vesicle-poor cells significantly increased in number, forming a huge meshwork overlying mature osteoblasts. Thus, ERpoor/vesicle-rich preosteoblasts appeared to respond well to PTH. We also attempted to unveil the cellular behavior of these preosteoblasts against PTH and to dissect the role of osteoclasts on the mediation of PTH anabolic actions. PTH stimulated the proliferation of ER-poor/vesicle-rich preosteoblasts and bone formation in mature osteoblasts. However, an increased population of ER-poor/vesicle-rich preosteoblasts appears to require cell coupling from osteoclasts to differentiate into ER-rich/vesicle-poor preosteoblasts and mature osteoblasts. Without osteoclasts, PTH could induce neither preosteoblastic differentiation into mature osteoblasts nor subsequent bone formation. In this mini-review, we will introduce preosteoblasts in vivo consisting of several cell types with different ultrastructural properties and PTH action on preosteoblasts

The Literary Text in a German-language Lesson, Methods for Working with Authentic Texts

Das Thema dieser Diplomarbeit heisst "Die Arbeit mit literarischen Texten. Methoden und Formen der Arbeit mit authentischen Texten". Die Arbeit bewegt sich sowohl im literaturtheoretischen als auch im didaktischen Bereich. Die Arbeit ist in einen theoretischen und zwei praktische Teile gegliedert. Ziel des theoretischen Teiles ist es, verschiedene Ansichten zum Thema zu sammeln und zu ordnen (die Problematik der literarischen Texte in pädagogischen Dokumenten, die Auswahl der literarischen Texte, die allgemeinen Prinzipien und Phasen der Arbeit mit literarischen und authentischen Texten, die Typologie der Übungen) und die zur Verfügung stehenden Methoden und Formen der Arbeit zu beschreiben.V teoretické části jsem na základě prostudované literatury vymezil pojmy literární text, autentický text a neautentický text. Pojem literární text užívám pro psaný text umělecký. Tento text chápu zároveň jako autentický, neboť literární text čteme zpravidla tak, jak byl napsán, což odpovídá významu autentického textu. Autentickým textem neliterárním rozumím texty funkčního stylu prostě sdělovacího (jízdenky, dopis, inzerát, novinový článek) a nikoliv uměleckého. Neautentickým textem zde rozumím text didaktizovaný, uměle vytvořený tvůrci učebnice pro potřeby vyučování - např. z důvodu prezentace nového gramatického jevu a nové slovní zásoby. Analýzou pedagogických dokumentů jsem dospěl k závěru, že práce s texty (včetně literárních) je v těchto dokumentech dostatečně zpracována. Dokumenty jsou učitelům dobrým návodem a pomocníkem. Diplomová práce zmapovala možnosti práce s literárním textem ve výuce včetně existujících pozitiv i možných obtíží. Dále byly představeny různé metodické postupy práce s literárními texty.Katedra germanistikyFaculty of EducationPedagogická fakult