Transporters, Ion Channels, and Junctional Proteins in Choroid Plexus Epithelial Cells.

The choroid plexus (CP) plays significant roles in secreting cerebrospinal fluid (CSF) and forming circadian rhythms. A monolayer of epithelial cells with tight and adherens junctions of CP forms the blood-CSF barrier to control the movement of substances between the blood and ventricles, as microvessels in the stroma of CP have fenestrations in endothelial cells. CP epithelial cells are equipped with several kinds of transporters and ion channels to transport nutrient substances and secrete CSF. In addition, junctional components also contribute to CSF production as well as blood-CSF barrier formation. However, it remains unclear how junctional components as well as transporters and ion channels contribute to the pathogenesis of neurodegenerative disorders. In this manuscript, recent findings regarding the distribution and significance of transporters, ion channels, and junctional proteins in CP epithelial cells are introduced, and how changes in expression of their epithelial proteins contribute to the pathophysiology of brain disorders are reviewed.

Biocompatibility of dental implants coated with hydroxyapatite using pulsed Er:YAG laser deposition.

We aimed to improve the biocompatibility and osteoinductive potential of Ti implants using a simulated intraoral hydroxyapatite (HAp) coating. We devised a novel surface treatment for aggressive induction of osteoblast adhesion and bone regeneration on the implant surface. A thin α-tricalcium phosphate (α-TCP) film was deposited on the implant surface using a pulsed Er:YAG laser. The coating was converted to HAp through artificial saliva immersion, which was confirmed using scanning electron microscopy (SEM) and X-ray diffraction (XRD). SEM showed needle-like HAp crystals on the Ti disks and sandblasted implant surfaces after immersion in artificial saliva for 96 h. Microcomputed tomography and histological evaluation 4 and 8 weeks after implantation into beagle dog mandibles showed that the HAp-coated implant was biocompatible and exhibited superior osteoinduction compared to that of sandblasted implants. Coating the implant surface with HAp using an Er:YAG laser has potential as a new method of the implant-surface debridement.

E-Cadherin Is Expressed in Epithelial Cells of the Choroid Plexus in Human and Mouse Brains.

Evidence showing the functional significance of the choroid plexus is accumulating. Epithelial cells with tight and adherens junctions of the choroid plexus play important roles in cerebrospinal fluid production and circadian rhythm formation. Although specific types of cadherin expressed in adherens junctions of choroid plexus epithelium (CPE) have been examined, they remained uncertain. Recent mass spectrometry and immunolocalization analysis revealed that non-epithelial cadherins, P- and N-cadherins, are expressed in the lateral membrane of CPE, whereas E-cadherin expression has not been confirmed in CPE of humans or mice. In this study, we examined E-cadherin expression in CPE of mice and humans by RT-PCR, immunohistochemical-, and Western blotting analyses. We confirmed, by using RT-PCR analysis, the mRNA expression of E-cadherin in the choroid plexus of mice. The immunohistochemical expression of E-cadherin was noted in the lateral membrane of CPE of mice and humans. We further confirmed, in Western blotting, the specific immunoreactivity for E-cadherin. Immunohistochemically, the expression of E- and N-cadherins or vimentin was unevenly distributed in some CPE, whereas that of E- and P-cadherins or ß-catenin frequently co-existed in other CPE. These findings indicate that E-cadherin is expressed in the lateral membrane of CPE, possibly correlated with the expression of other cadherins and cytoplasmic proteins.

Distribution of Monocarboxylate Transporters in Brain and Choroid Plexus Epithelium.

The choroid plexus (CP) plays central roles in regulating the microenvironment of the central nervous system by secreting the majority of cerebrospinal fluid (CSF) and controlling its composition. A monolayer of epithelial cells of CP plays a significant role in forming the blood-CSF barrier to restrict the movement of substances between the blood and ventricles. CP epithelial cells are equipped with transporters for glucose and lactate that are used as energy sources. There are many review papers on glucose transporters in CP epithelial cells. On the other hand, distribution of monocarboxylate transporters (MCTs) in CP epithelial cells has received less attention compared with glucose transporters. Some MCTs are known to transport lactate, pyruvate, and ketone bodies, whereas others transport thyroid hormones. Since CP epithelial cells have significant carrier functions as well as the barrier function, a decline in the expression and function of these transporters leads to a poor supply of thyroid hormones as well as lactate and can contribute to the process of age-associated brain impairment and pathophysiology of neurodegenerative diseases. In this review paper, recent findings regarding the distribution and significance of MCTs in the brain, especially in CP epithelial cells, are summarized.

Metabolites and Biomarker Compounds of Neurodegenerative Diseases in Cerebrospinal Fluid.

Despite recent advances in diagnostic procedures for neurological disorders, it is still difficult to definitively diagnose some neurodegenerative diseases without neuropathological examination of autopsied brain tissue. As pathological processes in the brain are frequently reflected in the components of cerebrospinal fluid (CSF), CSF samples are sometimes useful for diagnosis. After CSF is secreted from the choroid plexus epithelial cells in the ventricles, some flows in the brain, some is mixed with intracerebral interstitial fluid, and some is excreted through two major drainage pathways, i.e., the intravascular periarterial drainage pathway and the glymphatic system. Accordingly, substances produced by metabolic and pathological processes in the brain may be detectable in CSF. Many papers have reported changes in the concentration of substances in the CSF of patients with metabolic and neurological disorders, some of which can be useful biomarkers of the disorders. In this paper, we show the significance of glucose- and neurotransmitter-related CSF metabolites, considering their transporters in the choroid plexus; summarize the reported candidates of CSF biomarkers for neurodegenerative diseases, including amyloid-ß, tau, α-synuclein, microRNAs, and mitochondrial DNA; and evaluate their potential as efficient diagnostic tools.

Immunohistochemical expression of osteopontin and collagens in choroid plexus of human brains.

Evidence showing the functional significance of the choroid plexus is accumulating. Although it is clinically well-known that calcification is frequently seen in the choroid plexus of aged human brains, it is unclear why calcification occurs in the aged choroid plexus and what exert effects on the calcification has. In this study, immunohistochemical localizations of collagens and other molecules related to fibrosis or calcification were investigated on the choroid plexus of autopsied human brains. Densely fibrous or calcified materials were located in the stroma just below the epithelial cells of the choroid plexus of all human brains examined. Immunoreactivity for collagen type I was identified in the stroma just below the epithelial cells, consistent with the densely fibrous or calcified area, whereas that for collagen type III was observed in almost all stroma other than the densely fibrous or calcified areas. Linear or membranous immunoreactivity for collagen type IV was intermittently localized on the epithelium-facing side of the materials, suggesting an injured basement membrane. In addition, clear immunoreactivity for osteopontin was localized on the epithelium-facing side of the fibrous or calcified materials as well as in the cytoplasm of epithelial cells. These findings indicate that collagen type I exists in contact with osteopontin in and around the densely fibrous or calcified materials in the choroid plexus. They suggest that the densely fibrous or calcified materials are deposited in the subepithelial stroma just below an injured basement membrane of epithelial cells via the collagen type I and osteopontin.

Immunoreactivity of receptor and transporters for lactate located in astrocytes and epithelial cells of choroid plexus of human brain.

Glucose metabolism produces lactate and hydrogen ions in an anaerobic environment. Cerebral ischemia or hypoxia is believed to become progressively lactacidemic. Monocarboxylate transporters (MCTs) in endothelial cells are essential for the transport of lactate from the blood into the brain. In addition, it is considered that MCTs located in astrocytic and neuronal cells play a key role in the shuttling of energy metabolites between neurons and astrocytes. However, roles of lactate in the brain remain to be clarified. In this study, the localization of lactate transporters and a receptor for cellular uptake of lactate was immunohistochemically examined in autopsied human brains. Immunoreactivity for MCT1 was observed in the apical cytoplasmic membrane of some epithelial cells in the choroid plexus as well as astrocytes and the capillary wall, whereas that for MCT4 was found in the basolateral cytoplasmic membrane of small number of epithelial cells as well as astrocytes and the capillary wall. In addition, immunoreactivity for the hydroxy-carboxylic acid 1 receptor (HCA1 receptor), a receptor for cellular uptake of lactate, was also found on the basolateral cytoplasmic membrane of epithelial cells as well as astrocytic and neuronal cells. Immunoreactivity for lactate dehydrogenase (LDH)-B was observed in the cytoplasm of epithelial cells in the choroid plexus as well as astrocytes and the capillary wall. These immunohistochemical findings indicate the localization of MCT1, MCT4, the HCA1 receptor, and LDH-B in epithelial cells of the choroid plexus as well as astrocytes, and suggest the transport of intravascular lactate into the brain through epithelial cells of the choroid plexus as well as cerebral vessels and the possibility of lactate being utilized in epithelial cells.

Glucose, Fructose, and Urate Transporters in the Choroid Plexus Epithelium.

The choroid plexus plays a central role in the regulation of the microenvironment of the central nervous system by secreting the majority of the cerebrospinal fluid and controlling its composition, despite that it only represents approximately 1% of the total brain weight. In addition to a variety of transporter and channel proteins for solutes and water, the choroid plexus epithelial cells are equipped with glucose, fructose, and urate transporters that are used as energy sources or antioxidative neuroprotective substrates. This review focuses on the recent advances in the understanding of the transporters of the SLC2A and SLC5A families (GLUT1, SGLT2, GLUT5, GLUT8, and GLUT9), as well as on the urate-transporting URAT1 and BCRP/ABCG2, which are expressed in choroid plexus epithelial cells. The glucose, fructose, and urate transporters repertoire in the choroid plexus epithelium share similar features with the renal proximal tubular epithelium, although some of these transporters exhibit inversely polarized submembrane localization. Since choroid plexus epithelial cells have high energy demands for proper functioning, a decline in the expression and function of these transporters can contribute to the process of age-associated brain impairment and pathophysiology of neurodegenerative diseases.

Immunoreactivities for hepcidin, ferroportin, and hephaestin in astrocytes and choroid plexus epithelium of human brains.

Iron plays essential roles in the central nervous system. However, how the iron level is regulated in brain cells including glia and neurons remains to be fully clarified. In this study, the localizations of hepcidin, ferroportin, and hephaestin, which are known to be involved in iron efflux, were immunohistochemically examined in autopsied human brains. Immunoreactivities for hepcidin and ferroportin were observed in granular structures within the cytoplasm of reactive astrocytes and epithelial cells of the choroid plexus. Granular structures showing immunoreactivities for hepcidin and ferroportin were also stained with antibodies for early endosome antigen 1 (EEA1). In addition, immunoreactivity for hephaestin was observed in the cytoplasm of epithelial cells of the choroid plexus as well as reactive astrocytes. Immunoreactivity for hephaestin in the cytoplasm of reactive astrocytes was occasionally colocalized with immunoreactivity for EEA1, while that of hephaestin was frequently observed in the cytoplasm showing no immunoreactivity for EEA1. These findings suggest that immunoreactivities for hepcidin and ferroportin are localized in close proximity to granular structures showing immunoreactivity for EEA1 in the cytoplasm of human brain astrocytes. They also suggest that immunoreactivity of hephaestin is localized in the cytoplasm of the choroid plexus epithelium as well as reactive astrocytes of human brains.

Disturbance of Intracerebral Fluid Clearance and Blood-Brain Barrier in Vascular Cognitive Impairment.

The entry of blood-borne macromolecular substances into the brain parenchyma from cerebral vessels is blocked by the blood-brain barrier (BBB) function. Accordingly, increased permeability of the vessels induced by insult noted in patients suffering from vascular dementia likely contributes to the cognitive impairment. On the other hand, blood-borne substances can enter extracellular spaces of the brain via endothelial cells at specific sites without the BBB, and can move to brain parenchyma, such as the hippocampus and periventricular areas, adjacent to specific sites, indicating the contribution of increased permeability of vessels in the specific sites to brain function. It is necessary to consider influx and efflux of interstitial fluid (ISF) and cerebrospinal fluid (CSF) in considering effects of brain transfer of intravascular substances on brain function. Two pathways of ISF and CSF are recently being established. One is the intramural peri-arterial drainage (IPAD) pathway of ISF. The other is the glymphatic system of CSF. Dysfunction of the two pathways could also contribute to brain dysfunction. We review the effects of several kinds of insult on vascular permeability and the failure of fluid clearance on the brain function.

Epigallocatechin Gallate-Modified Gelatin Sponges Treated by Vacuum Heating as a Novel Scaffold for Bone Tissue Engineering.

Chemical modification of gelatin using epigallocatechin gallate (EGCG) promotes bone formation in vivo. However, further improvements are required to increase the mechanical strength and bone-forming ability of fabricated EGCG-modified gelatin sponges (EGCG-GS) for practical applications in regenerative therapy. In the present study, we investigated whether vacuum heating-induced dehydrothermal cross-linking of EGCG-GS enhances bone formation in critical-sized rat calvarial defects. The bone-forming ability of vacuum-heated EGCG-GS (vhEGCG-GS) and other sponges was evaluated by micro-computed tomography and histological staining. The degradation of sponges was assessed using protein assays, and cell morphology and proliferation were verified by scanning electron microscopy and immunostaining using osteoblastic UMR106 cells in vitro. Four weeks after the implantation of sponges, greater bone formation was detected for vhEGCG-GS than for EGCG-GS or vacuum-heated gelatin sponges (dehydrothermal cross-linked sponges without EGCG). In vitro experiments revealed that the relatively low degradability of vhEGCG-GS supports cell attachment, proliferation, and cell-cell communication on the matrix. These findings suggest that vacuum heating enhanced the bone forming ability of EGCG-GS, possibly via the dehydrothermal cross-linking of EGCG-GS, which provides a scaffold for cells, and by maintaining the pharmacological effect of EGCG.

Immunoreactivity of urate transporters, GLUT9 and URAT1, is located in epithelial cells of the choroid plexus of human brains.

It has been suggested that urate plays a protective role in neurons, while hyperuricemia is correlated with atherosclerosis and cardiovascular disease. However, whether there is a system that directly transports urate into the brain remains to be clarified. In this study, the localization of glucose transporter 9 (GLUT9) and urate transporter 1 (URAT1), which are known to be representative reabsorptive urate transporters, was immunohistochemically examined in autopsied human brains. Immunoreactivity of GLUT9 was observed on the apical side of the cytoplasm of epithelial cells in the choroid plexus and in the cilia of ependymal cells of the human brain. Immunoreactivity of URAT1 was observed on the basolateral side of the cytoplasm of epithelial cells in the choroid plexus. In addition, immunoreactivity of GLUT9 and URAT1 was not observed in microvessels of the human brains. The choroid plexus and renal proximal tubule were similar in having a polarized distribution of these two transporters with the two transporters on opposite membranes, but the two transporters' distribution differs between the choroid plexus and the kidney in terms of which membrane (apical/basal) expresses which transporter. These findings support the hypothesis of the direct transport of intravascular urate into the central nervous system through the choroid plexus.

In vivo bioactivity of porous polyetheretherketone with a foamed surface.

The in vivo bioactivity of porous polyetheretherketone (PEEK) with a foamed surface was evaluated using rabbit femoral bone. Cylindrical porous PEEK scaffolds, with pore diameter of 550 µm and porosity of 70%, were first prepared and immersed in 98% sulfuric acid, and then washed and immersed in 3 M potassium carbonate solution used as a foaming reagent. Numerous open pores of various sizes, as well as new functional groups, were visualized on the treated PEEK surface by scanning electron microscopy and X-ray photoelectron spectroscopy, respectively. Micro computed tomography (micro-CT) showed that the volumetric density of treated PEEK was higher than that of bare PEEK at 8 weeks after surgery (p<0.05). Additionally, von Kossa staining indicated ingrowth of mature new bone tissue at 4 weeks relative to the bare PEEK group. Our data indicate that surface-treated PEEK exhibited improved bioactivity in vivo.

Glucose transporter 8 immunoreactivity in astrocytic and microglial cells in subependymal areas of human brains.

Glucose transporter 8 (GLUT8), a glucose/fructose transporter, has been shown to be expressed in neuronal cells in several brain areas. A recent immunohistochemical study has shown the presence of GLUT8 in the cytoplasm of epithelial cells of the choroid plexus and ependymal cells. In this study, localization of GLUT8 in glial cells was investigated using immunohistochemical methods. Immunoreactivity for GLUT8 was observed in cells showing astrocytic or microglial structural features located around the lateral ventricles. Confocal microscopic examination revealed that subependymal GLUT8-positive cells with large amounts of cytoplasm mainly show clear immunoreactivity for vimentin, while they were also colocalized with weak immunoreactivity for glial fibrillary acidic protein (GFAP) within the cytoplasm of some cells. In addition, some GLUT8-positive cells with small amounts of cytoplasm and small nuclei showed CD68 or HLA-DR immunoreactivity, indicating them to be cells of microglia/macrophage lineage. These findings suggest that glucose/fructose is transported into the cytoplasm of vimentin- or GFAP-positive astrocytic and CD68- or HLA-DR-positive microglial cells located around the lateral ventricle.

Evaluation of the clinical and aesthetic outcomes of Straumann(®) Standard Plus implants supported single crowns placed in non-augmented healed sites in the anterior maxilla: a 5-8 years retrospective study.

PURPOSE: To evaluate the long-term aesthetic outcome of the single crowns supported by soft tissue level implants placed in healed sites in the anterior maxilla region via the pink aesthetic score (PES) and the white aesthetic score (WES). MATERIAL AND METHODS: According to the inclusion criteria, patients who had received a single Straumann(®) Standard Plus implant in the anterior maxilla at the Shanghai 9th People's Hospital between 2005 and 2008 were invited for a re-examination based on a number of inclusion criteria and exclusion criteria. Clinical, radiographic and aesthetic outcomes (PES/WES) were assessed during their revisit at 5-8 years after crown placement. RESULTS: Forty-five patients were enrolled in the study. All 45 implants were successfully integrated and most of the implants did not show signs of peri-implant disease at the time of the assessment. The marginal bone resorption was 1.10 ± 0.92 mm. The mean total PES was 8.48 ± 2.62 at the baseline, 9.57 ± 2.37 at the 6-10 months revisit and 9.01 ± 2.45 at the 5-8 years follow-up. The scores of the mesial and distal papillae increased significantly between the baseline and 6-10 months follow-up, this improvement remained stable at the 5-8 years follow-up. The scores of soft tissue level, colour of the soft tissue, soft tissue texture and the alveolar process decreased significantly between the 6-10 months and 5-8 years revisits. The mean WES was 7.83 ± 1.60 at the baseline and 7.72 ± 1.43 at the 5-8 years revisit. There was no significant difference of the WES between the baseline and 5-8 years revisit. CONCLUSION: The possibility of spontaneous papillae regeneration after implant treatment and the long-term stability of the regenerated papillae were confirmed. However, recession of the facial soft tissue has been found. The incidence of the recession at thin biotype sites tended to be higher.

Case of vascular air embolism during holmium laser enucleation of the prostate.

Vascular air embolism is a rare complication during transurethral surgery. A case of air embolism during holmium laser enucleation of the prostate in a 76-year-old man is presented. During the step of morcellation, the patient's blood pressure suddenly oscillated up and down, and end-tidal CO2 and arterial saturation decreased. Transesophageal and transthoracic echocardiography showed air collection in the right atrium. It was also discovered that incorrect assembly of the tube from the morcellator caused rapid entrainment of air into the vein. Computed tomography and abdominal X-ray showed niveau formation in the femoral vein and air collection in the pelvic retroperitoneal space. The patient recovered with careful observation and was discharged 7 days after the operation with no sequelae. This report is presented to remind urologists of this unusual complication that can occur during holmium laser enucleation of the prostate procedures.

Incidence of intussusception as studied from a hospital-based retrospective survey over a 10-year period (2001-2010) in Akita Prefecture, Japan.

One concern about rotavirus vaccines is its possible association with intussusception. Thus, it is necessary to determine the baseline incidence for intussusception in the first year of life in places where rotavirus vaccines are introduced. However, few safety data exist for the period at which the first dose of Rotarix and RotaTeq are allowed to administer in Japan. The first dose of Rotarix is scheduled to administer at 6-20 weeks of age and that of RotaTeq is scheduled to administer at 6-24 weeks of age; the upper limits for these vaccines is later than the upper limit recommended by the World Health Organization by 5 and 9 weeks, respectively. We performed a retrospective cross-sectional study by reviewing medical charts of all hospitals that provided pediatric beds in Akita Prefecture, Japan, and identifying the cases of intussusception that met the Brighton criteria level 1 in these hospitals between January 2001 and December 2010. During this 10-year period, 122 children younger than 1 year of age were diagnosed with intussusception. The incidence of intussusception was estimated at 158 per 100,000 person-years among children younger than 1 year (95% confidence interval, 131-188), 10 per 100,000 person-years for children aged 0-2 months, 165 for children aged 3-5 months, and 300 for children aged 6-8 months. This rapid and substantial increase in the incidence of intussusception during the first year of life should be considered when formulating the immunization schedule for administering rotavirus vaccines in Japan.

Study on motor learning of sternocleidomastoid muscles during ballistic voluntary opening.

PURPOSE: Neck pain is one of the main symptoms of temporomandibular disorder. Muscle activity of the sternocleidomastoid muscle during occlusion has been clarified in recent years. We reported that when healthy individuals were instructed to chew rapidly, the activity of the sternocleidomastoid muscle responded to activity of the masseter muscle, however, during voluntary jaw opening, activity of the sternocleidomastoid muscle did not respond, but worked actively due to motor programming. The objective of the present study was to investigate the learning effects of repetitive training, that is, changes in activity mode of the neuromuscular system. MATERIALS AND METHODS: The sternocleidomastoid and the anterior belly of digastric muscles in 8 healthy male adults were analyzed. In response to acoustic stimulation, each subject was instructed to open their mouth as quickly and widely as possible a total of 30 times with a break between measurements. EMG-reaction times (RT) of the sternocleidomastoid and anterior belly of digastric muscles were measured, and the length of time from the start of EMG activity of agonist to the start of actual movement was measured. RESULTS: In all subjects, at first measurement, EMGRT of the sternocleidomastoid muscle did not precede that of the anterior belly of digastric muscle. With each measurement, the difference in EMG-RT between the sternocleidomastoid and the anterior belly of digastric muscles decreased, and in 6 of the 8 subjects, EMG-RT of the sternocleidomastoid muscle preceded that of the anterior belly of digastric muscle. CONCLUSION: Repetitive task movement alters the start times of muscular activities, and from the perspective of EMG kinesiology, motor learning effects were confirmed with maximum ballistic voluntary jaw opening.