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Introduction Lateral radiography of the knee joint is frequently performed; however, the retake rate is high owing to positioning errors. Therefore, in this study, to reduce the required number and time of image retakes, we developed a system that can classify the tilting directions of lateral knee radiographs and evaluated the accuracy of the proposed method. Methods Using our system, the tilting directions of a lateral knee radiographs were classified into four direction categories. The system was developed by training the DCNN based on 50 cases of Raysum images and tested on three types test dataset; ten more cases of Raysum images, one case of flexed knee joint phantom images and 14 rejected knee joint radiographs. To train a deep convolutional neural network (DCNN), we employed Raysum images created via three-dimensional (3D) X-ray computed tomography (CT); 11 520 Raysum images were created from 60 cases of 3D CT data by changing the projection angles. Thereby, we obtained pseudo images attached with correct labels that are essential for training. Results The overall accuracy on each test dataset was 88.5 ± 7.0% (mean ± standard deviation), 81.4 ± 11.2%, and 73.3 ± 9.2%. The larger the tilting degree of the knee joint, the higher the classification accuracy. Conclusion DCNN could classify the tilting directions of a knee joint from lateral knee radiographs. Using Raysum images made it possible to facilitate creating dataset for training DCNN. The possibility was indicated for using support system of lateral knee radiographs. Implications for practice The system may also reduce the burden on patients and increase the work efficiency of radiological technologists.

Aim The present study aimed to propose a structural model to explain the interaction of physical, cognitive and social domains of health status in the incidence of disability in community-dwelling people aged ≥75 years. Methods We analyzed 185 older adults (mean age 79.4 years, 58.4% female) who participated in a baseline assessment from 2012 to 2013. They were followed for incident certification of care needs in the national long-term care insurance certification system during the 2 years. Baseline assessments included several measurements related to the physical, cognitive and social domains of health status. We compared the model fit index between two hypothesis models - the parallel model and the hierarchical model - using structural equation modeling. Results During the follow-up period, 15 participants (8.1%) were newly certified as requiring personal support from the long-term care insurance system. The structural equation modeling showed that the hierarchical model, indicating that cognitive and social status were indirectly associated with disability through physical status, had a better fit with the data than the parallel model, indicating that physical, cognitive and social status each were directly associated with disability. Conclusions The present results suggest that cognitive and social status might indirectly affect disability incidence through physical aging. Further research is required to examine the temporal relationship between physical, cognitive and social change using data over several time-periods. Geriatr Gerontol Int 2018; 18: 1614-1619.

Background Long-term prognosis of cancer patients has been improved along with the progress in chemotherapies. However, chemotherapy-related cardiac dysfunction (CTRCD) is emerging as a serious adverse effect as it worsens patients' outcome and quality of life. Thus, early detection of subclinical CTRCD is an important emerging issue in the management of cancer patients. Cardiac magnetic resonance (CMR) utilizes parametric mapping approach and strain analysis to provide detailed information about cardiac tissue and diastolic cardiac function. Purpose We examined whether the novel CMR imaging techniques are useful for early detection of CTRCD. Methods and results We performed both retrospective and prospective studies. (1) Retrospective study: We retrospectively enrolled 52 cancer patients (mean age 55.6±13 yrs., M/F=14/38) who had been treated with anthracyclines. We examined the usefulness of CMR for quantitative assessment of myocardial fibrosis caused by chemotherapies. We found that native T1 value was significantly prolonged in cancer patients compared with healthy controls (N=10) (1,279±56 vs. 1,240±34 msec, P=0.036). (2) Prospective study: A total of 99 consecutive female patients with breast cancer treated with chemotherapies were enrolled in this study from August 2017 to January 2019. To evaluate CTRCD in those patients, we performed CMR (at baseline and/or 6 months) and biomarkers analysis for cardiac troponin T (cTnT) and BNP at baseline and every 3 months during chemotherapies. In the 99 patients, 52 (mean age 53.0±12.7 yrs.) completed cardiac assessment at 6 months, and 6 (12%) developed CTRCD defined as a reduction in left ventricular ejection fraction (LVEF) >10% from baseline and below 53% without symptoms. In patients with CTRCD (CTRCD group, N=6), as compared with those without it (non-CTRCD group, N=46), native T1 value was significantly prolonged after chemotherapies (1,303±32 vs. 1,322±22 msec at 6 months, P=0.03). Plasma cTnT levels at 3 months were also significantly higher in the CTRCD group compared with the non-CTRCD group [0.022 (IQR 0.015–0.026) vs. 0.01 (0.006–0.014) ng/mL, P=0.024], whereas there was no difference in BNP values. In the 52 patients, 28 (mean age 56.3±12.3 yrs.) underwent CMR both before and 6 months after chemotherapies. In those patients, LVEF and global radial strain were significantly decreased at 6 months from baseline (LVEF, from 70.5±4.6 to 66.0±7.1%; global radial strain, from 70.0±22.5 to 61.1±22.6%, respectively, both P Conclusions These results indicate that novel CMR imaging techniques are useful for early detection of CTRCD among cancer patients treated with chemotherapies.

Spike conduction velocity characteristically differs between myelinated and unmyelinated axons. Here we test whether spikes of myelinated and unmyelinated paths differ in other respects by measuring rat retinal ganglion cell (RGC) spike duration in the intraretinal, unmyelinated nerve fiber layer and the extraretinal, myelinated optic nerve and optic chiasm. We find that rapid spike firing and illumination broaden spikes in intraretinal axons but not in extraretinal axons. RGC axons thus initiate spikes intraretinally and normalize spike duration extraretinally. Additionally, we analyze spikes that were recorded in a previous study of rhesus macaque retinogeniculate transmission and find that rapid spike firing does not broaden spikes in optic tract. The spike normalization we find reduces the number of spike properties that can change during RGC light responses. However, this is not because identical spikes fire in all axons. Instead, our recordings show that different subtypes of RGC generate axonal spikes of different durations and that the differences resemble spike duration increases that alter neurotransmitter release from other neurons. Moreover, previous studies have shown that RGC spikes of shorter duration can fire at higher maximum frequencies. These properties should facilitate signal transfer by different mechanisms at RGC synapses onto subcortical target neurons.

Repeated spike firing can transmit information at synapses and modulate spike timing, shape, and conduction velocity. These latter effects have been found to result from voltage-induced changes in ion currents and could alter the signals carried by axons. Here, we test whether Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) regulates spike propagation in adult rat optic nerve. We find that small-, medium-, and large-diameter axons bind anti-Thr286-phosphorylated CaMKII (pT286) antibodies and that, in isolated optic nerves, electrical stimulation reduces pT286 levels, spike propagation is hastened by CaMKII autophosphorylation and slowed by CaMKII dephosphorylation, single and multiple spikes slow propagation of subsequently activated spikes, and more frequent stimulation produces greater slowing. Likewise, exposing freely moving animals to flickering illumination reduces pT286 levels in optic nerves and electrically eliciting spikes in vivo in either the optic nerve or optic chiasm slows subsequent spike propagation in the optic nerve. By increasing the time that elapses between successive spikes as they propagate, pT286 dephosphorylation and activity-induced spike slowing reduce the frequency of propagated spikes below the frequency at which they were elicited and would thus limit the frequency at which axons synaptically drive target neurons. Consistent with this, the ability of retinal ganglion cells to drive at least some lateral geniculate neurons has been found to increase when presented with light flashes at low and moderate temporal frequencies but less so at high frequencies. Activity-induced decreases in spike frequency may also reduce the energy required to maintain normal intracellular Na(+) and Ca(2+) levels. SIGNIFICANCE STATEMENT By propagating along axons at constant velocities, spikes could drive synapses as frequently as they are initiated. However, the onset of spiking has been found to alter the conduction velocity of subsequent (“follower”) spikes in various preparations. Here, we find that spikes reduce spike frequency in rat optic nerve by slowing follower spike propagation and that electrically stimulated spiking ex vivo and spike-generating flickering illumination in vivo produce net decreases in axonal Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) autophosphorylation. Consistent with these effects, propagation speed increases and decreases, respectively, with CaMKII autophosphorylation and dephosphorylation. Lowering spike frequency by CaMKII dephosphorylation is a novel consequence of axonal spiking and light adaptation that could decrease synaptic gain as stimulus frequency increases and may also reduce energy use.

Immunohistochemical and ex vivo anatomical studies have provided many glimpses of the variety, distribution, and signaling components of vertebrate retinal neurons. The beauty of numerous images published to date, and the qualitative and quantitative information they provide, indicate that these approaches are fundamentally useful. However, obtaining these images entailed tissue handling and exposure to chemical solutions that differ from normal extracellular fluid in composition, temperature, and osmolarity. Because the differences are large enough to alter intercellular and intracellular signaling in neurons, and because retinae are susceptible to crush, shear, and fray, it is natural to wonder if immunohistochemical and anatomical methods disturb or damage the cells they are designed to examine. Tissue fixation is typically incorporated to guard against this damage and is therefore critically important to the quality and significance of the harvested data. Here, we describe mechanisms of fixation; advantages and disadvantages of using formaldehyde and glutaraldehyde as fixatives during immunohistochemistry; and modifications of widely used protocols that have recently been found to improve cell shape preservation and immunostaining patterns, especially in proximal retinal neurons.

[Purpose] The purpose of this study was to examine the associations between the settings of exercise habits and health-related outcomes in community-dwelling older adults. [Subjects] A total of 304 Japanese community-dwelling older adults (70.3 ± 4.1 years; 113 males and 191 females) participated in this study. [Methods] Demographic characteristics, medical conditions, exercise habits, and health-related outcomes were assessed by face-to-face interviews and self-reported questionnaires. Older adults who had exercise habits were classified into two groups: individual- and group-based exercise habits groups, and the health-related outcomes were compared between groups. [Results] The scores for the Geriatric Depression Scale, exercise self-efficacy, and dietary variety of older adults who had group-based exercise habits were better than those of older adults who had individual-based exercise habits. In addition, the exercise settings (individual- and group-based) were significantly associated with scores for the Geriatric Depression Scale (odds ratio = 0.76) and exercise self-efficacy (odds ratio = 1.26), even after adjusting for age and gender. [Conclusion] These results implied that habitual exercise in group settings may have an effective role in promoting exercise self-efficacy and mental health.

Dopamine- and tyrosine hydroxylase-immunopositive cells (TH cells) modulate visually driven signals as they flow through retinal photoreceptor, bipolar, and ganglion cells. Previous studies suggested that TH cells release dopamine from varicose axons arborizing in the inner and outer plexiform layers after glutamatergic synapses depolarize TH cell dendrites in the inner plexiform layer and these depolarizations propagate to the varicosities. Although it has been proposed that these excitatory synapses are formed onto appendages resembling dendritic spines, spines have not been found on TH cells of most species examined to date or on TH cell somata that release dopamine when exposed to glutamate receptor agonists. By use of protocols that preserve proximal retinal neuron morphology, we have examined the shape, distribution, and synapse-related immunoreactivity of adult rat TH cells. We report here that TH cell somata, tapering and varicose inner plexiform layer neurites, and varicose outer plexiform layer neurites all bear spines, that some of these spines are immunopositive for glutamate receptor and postsynaptic density proteins (viz., GluR1, GluR4, NR1, PSD-95, and PSD-93), that TH cell somata and tapering neurites are also immunopositive for a γ-aminobutyric acid (GABA) receptor subunit (GABAA Rα1 ), and that a synaptic ribbon-specific protein (RIBEYE) is found adjacent to some colocalizations of GluR1 and TH in the inner plexiform layer. These results identify previously undescribed sites at which glutamatergic and GABAergic inputs may stimulate and inhibit dopamine release, especially at somata and along varicose neurites that emerge from these somata and arborize in various levels of the retina. J. Comp. Neurol. 525:1707-1730, 2017. © 2016 Wiley Periodicals, Inc.

Protocols for characterizing cellular phenotypes commonly use chemical fixatives to preserve anatomical features, mechanically stabilize tissue, and stop physiological responses. Formaldehyde, diluted in either phosphate-buffered saline or phosphate buffer, has been widely used in studies of neurons, especially in conjunction with dyes and antibodies. However, previous studies have found that these fixatives induce the formation of bead-like varicosities in the dendrites and axons of brain and spinal cord neurons. We report here that these formaldehyde formulations can induce bead formation in the dendrites and axons of adult rat and rabbit retinal ganglion cells, and that retinal ganglion cells differ from hippocampal, cortical, cerebellar, and spinal cord neurons in that bead formation is not blocked by glutamate receptor antagonists, a voltage-gated Na(+) channel toxin, extracellular Ca(2+) ion exclusion, or temperature shifts. Moreover, we describe a modification of formaldehyde-based fixatives that prevents bead formation in retinal ganglion cells visualized by green fluorescent protein expression and by immunohistochemistry.

Background Immune-checkpoint inhibitors (ICI) have recently been used in many types of cancers including non-small cell lung cancer (NSCLC). Interstitial lung disease associated with ICI (ICI-ILD) is a serious immune-related adverse event. However, comprehensive clinical characterization of ICI-ILD is still lacking. Methods We retrospectively analyzed clinical and imaging data of all consecutive ICI treated patients in Kurashiki Central Hospital (Okayama, Japan) between January 2016 and March 2019. We elucidated clinical characteristics, radiopathological patterns, therapeutic strategies and outcomes of ICI-ILD and analyzed the prognostic factor of ICI-ILD. Results Among 316 (222 NSCLC and 94 non-NSCLC) treated with ICI, 35 (11.1%) were diagnosed as ICI-ILD. Of those, NSCLC/gastric cancer/urinary cancer/head and neck cancer/malignant lymphoma were 26/2/2/4/1, respectively: mean age was 69.3 ± 9.0 years; 29 (82.9%) were men; 19 (54.3%) received Nivolumab monotherapy and 10 (28.6%) received Pembrolizumab monotherapy; Best objective response rate until ICI-ILD diagnosis was 48.6% (11/35); The median time (range) from ICI introduction to ICI-ILD was 57 (22-155) days. CTCAE grade 1/2/3 + 4/5 was 10 (28.6%)/14 (40.0%)/7 (20.0%)/4 (11.4%), respectively; Organizing pneumonia was found in 57.1% (20/35) in CT and 66.7% (4/6) in transbronchial lung biopsy; Lymphocyte differential count of bronchoalveolar lavage fluid (range) was 41 (33-60) %; 26 (74.3%) were treated with corticosteroid; 29 (82.9%) had ICI-ILD improved but 6 (17.1%) did not. The median survival time was significantly different between ILD improved group and non-improved group (17.1 months vs 0.8 months, p Conclusions In our real-world experience, half of ICI-ILD patients had a response toward malignancy. Organizing pneumonia was the most common radiopathological pattern and lymphocyte differential count of bronchoalveolar lavage fluid was elevated in ICI-ILD. If ICI-ILD does not improve, its prognosis may be very poor. Legal entity responsible for the study The authors. Funding Has not received any funding. Disclosure All authors have declared no conflicts of interest.

Esophagectomy, one of the most invasive of all gastrointestinal operations, is associated with a high frequency of postoperative complications and in-hospital mortality. The purpose of the present study was to determine whether exposure to the atomic bomb explosion at Hiroshima in 1945 might be a preoperative risk factor for in-hospital mortality after esophagectomy in esophageal cancer patients. We thus reviewed the outcomes of esophagectomy in 31 atomic bomb survivors with esophageal cancer and 96 controls (also with cancer but without atomic bomb exposure). We compared the incidences of postoperative complications and in-hospital mortality. Of the clinicopathological features studied, mean patient age was significantly higher in atomic bomb survivors than in controls. Of the postoperative complications noted, atomic bomb survivors experienced a longer mean period of endotracheal intubation and higher incidences of severe pulmonary complications, severe anastomotic leakage, and surgical site infection. The factors associated with in-hospital mortality were exposure to the atomic bomb explosion, pulmonary comorbidities, and electrocardiographic abnormalities. Multivariate analysis revealed that exposure to the atomic bomb explosion was an independent significant preoperative risk factor for in-hospital mortality. Exposure to the atomic bomb explosion is thus a preoperative risk factor for in-hospital death after esophagectomy to treat esophageal cancer.

Aims Ubiquilin-1 acts as an adaptor protein that mediates the translocation of polyubiquitinated proteins to the proteasome for degradation. Although previous studies suggested a key role of ubiquilin-1 in the pathogenesis of Alzheimer's disease (AD), a direct relationship between ubiquilin-1 and Hirano bodies in AD brains remains unknown. Methods By immunohistochemistry, we studied ubiquilin-1 and ubiquilin-2 expression in the frontal cortex and the hippocampus of six AD and 13 control cases. Results Numerous Hirano bodies, accumulated in the hippocampal CA1 region of AD brains, expressed intense immunoreactivity for ubiquilin-1. They were much less frequently found in control brains. However, Hirano bodies did not express a panel of markers for proteasome, autophagosome or pathogenic proteins, such as ubiquilin-2, ubiquitin, p62, LC3, beclin-1, HDAC6, paired helical filament (PHF)-tau, protein-disulphide isomerase (PDI) and phosphorylated TDP-43, but some of them expressed C9orf72. Ubiquilin-1-immunoreactive deposits were classified into four distinct morphologies, such as rod-shaped structures characteristic of Hirano bodies, dystrophic neurites contacting senile plaques, fragmented structures accumulated in the lesions affected with severe neuronal loss, and thread-shaped structures located mainly in the molecular layer of the hippocampus. Conclusions Ubiquilin-1 immunoreactivity is concentrated on Hirano bodies and dystrophic neurites in AD brains, suggesting that aberrant expression of ubiquilin-1 serves as one of pathological hallmarks of AD.

J. Satoh, H. Tabunoki, T. Ishida, Y. Saito and K. Arima (2013) Neuropathology and Applied Neurobiology39, 109–120 Accumulation of a repulsive axonal guidance molecule RGMa in amyloid plaques: a possible hallmark of regenerative failure in Alzheimer's disease brains Aims: RGMa is a repulsive guidance molecule that induces the collapse of axonal growth cones by interacting with the receptor neogenin in the central nervous system during development. It remains unknown whether RGMa plays a role in the neurodegenerative process of Alzheimer's disease (AD). We hypothesize that RGMa, if it is concentrated on amyloid plaques, might contribute to a regenerative failure of degenerating axons in AD brains. Methods: By immunohistochemistry, we studied RGMa and neogenin (NEO1) expression in the frontal cortex and the hippocampus of 6 AD and 12 control cases. The levels of RGMa expression were determined by qRT-PCR and Western blot in cultured human astrocytes following exposure to cytokines and amyloid beta (Aβ) peptides. Results: In AD brains, an intense RGMa immunoreactivity was identified on amyloid plaques and in the glial scar. In the control brains, the glial scar and vascular foot processes of astrocytes expressed RGMa immunoreactivity, while oligodendrocytes and microglia were negative for RGMa. In AD brains, a small subset of amyloid plaques expressed a weak NEO1 immunoreactivity, while some reactive astrocytes in both AD and control brains showed an intense NEO1 immunoreactivity. In human astrocytes, transforming growth factor beta-1 (TGFβ1), Aβ1–40 or Aβ1–42 markedly elevated the levels of RGMa, and TGFβ1 also increased its own levels. Coimmunoprecipitation analysis validated the molecular interaction between RGMa and the C-terminal fragment β of amyloid beta precursor protein (APP). Furthermore, recombinant RGMa protein interacted with amyloid plaques in situ. Conclusions: RGMa, produced by TGFβ-activated astrocytes and accumulated in amyloid plaques and the glial scar, could contribute to the regenerative failure of degenerating axons in AD brains.

Background: Body composition is an important component of health related fitness. Near-infrared spectroscopy (NIRS) is a non-invasive, simple and rapid method of assessing body fat percentage. However, it is unknown whether NIRS can accurately estimate FFM in community-dwelling frail elderly. Objectives: This study aimed to compare NIRS with bioelectrical impedance analysis (BIA) in FFM measurement. Design: Cross-sectional study. Setting: Shizuoka, Japan. Participants: The study population comprised 53 community-dwelling frail elderly (15 men, 38 women; mean age 84.8±6.4 years; body mass index 19.7±3.5 kg/m2). Measurement: FFM and percentage fat mass (%FM) were estimated using a NIRS device at two sites (biceps and calf) and compared to body composition measured by BIA. Simple linear regression and Bland–Altman analyses were used to determine agreement between the methods. Results: FFM determined by BIA highly correlated with that determined by NIRS at both the biceps and calf (r=0.92 for both; p

J. Satoh, H. Tabunoki, T. Ishida, Y. Saito and K. Arima (2012) Neuropathology and Applied Neurobiology38, 132–141 Immunohistochemical characterization of γ-secretase activating protein expression in Alzheimer's disease brains Aims: A recent study showed that γ-secretase activating protein (GSAP), derived from a C-terminal fragment of pigeon homolog (PION), increases amyloid-β (Aβ) production by interacting with presenilin-1 (PS1) and the β-secretase-cleaved C-terminal fragment of amyloid precursor protein (APP-CTF). In the study, knockdown of GSAP reduces production of Aβ and plaque formation in the brain of APPswe and PS1ΔE9 double transgenic mice without affecting the Notch-dependent pathway. Therefore, GSAP is an ideal target for designing γ-secretase modulators with least side effects in Alzheimer's disease (AD). However, at present, the precise distribution of GSAP in AD brains remains to be characterized. Methods: By immunohistochemistry, we studied GSAP expression in the frontal cortex and the hippocampus of 11 aged AD and 17 age-matched control cases. Results: GSAP immunoreactivity exhibited distinct morphological features, such as fine granular cytoplasmic deposits, dense nodular and patchy deposits, beads and string-like deposits, and diffuse dot-like deposits. In both AD and control brains, a fairly small subset of cerebral cortical and hippocampal neurones expressed fine granular cytoplasmic deposits, while diffuse dot-like deposits were more frequently found in the neuropil and neuronal processes, particularly enriched in the hippocampal CA2 and CA3 regions. Among GSAP-immunoreactive deposits, dense nodular and patchy deposits, located in the neuropil and closely associated with PS1 expression and Aβ deposition, indicated the most distinguishing features of AD pathology. Conclusions: Aberrant regulation of GSAP expression plays a key role in acceleration of γ-cleavage of APP-CTF and accumulation of Aβ in AD brains.

Warfarin, a coumarin rodenticide, is commonly used worldwide for rodent control, and is often reported as the cause for poisoning accidents in nontarget animals, in particular bird species. However, the metabolism of warfarin in birds is still unclear. In a previous study, we found an unknown warfarin metabolite in chicken cytosolic fractions. In the present study, we aimed to clarify the cytosolic warfarin metabolites in chickens compared with those in rats. The cytosol fractions of both chicken and rat livers showed the metabolic activity of 2 diastereomers and 2 enantiomers of warfarin alcohol. In chicken cytosol, we found that the production level of (S)-warfarin-(S)-alcohol was markedly higher (32-fold) than that in rat cytosol. From the results of the inhibition assay, we finally suggest that aldehyde oxidase may mainly contribute to the warfarin alcohol products in chicken cytosol.

The current-passing pore of mammalian hyperpolarization-activated, cyclic nucleotide-gated (HCN) channels is formed by subunit isoforms denoted HCN1–4. In various brain areas, antibodies directed against multiple isoforms bind to single neurons, and the current (Ih) passed during hyperpolarizations differs from that of heterologously expressed homomeric channels. By contrast, retinal rod, cone, and bipolar cells appear to use homomeric HCN channels. Here, we assess the generality of this pattern by examining HCN1 and HCN4 immunoreactivity in rat retinal ganglion cells, measuring Ih in dissociated cells, and testing whether HCN1 and HCN4 proteins coimmunoprecipitate. Nearly half of the ganglion cells in whole-mounted retinae bound antibodies against both isoforms. Consistent with colocalization and physical association, 8-bromo-cAMP shifted the voltage sensitivity of Ih less than that of HCN4 channels and more than that of HCN1 channels, and HCN1 coimmunoprecipitated with HCN4 from membrane fraction proteins. Finally, the immunopositive somata ranged in diameter from the smallest to the largest in rat retina, the dendrites of immunopositive cells arborized at various levels of the inner plexiform layer and over fields of different diameters, and Ih activated with similar kinetics and proportions of fast and slow components in small, medium, and large somata. These results show that different HCN subunits colocalize in single retinal ganglion cells, identify a subunit that can reconcile native Ih properties with the previously reported presence of HCN4 in these cells, and indicate that Ih is biophysically similar in morphologically diverse retinal ganglion cells and differs from Ih in rods, cones, and bipolar cells. J. Comp. Neurol. 519:2546–2573, 2011. © 2011 Wiley-Liss, Inc.

Recent evidence has shown that d-aspartate modulates hormone secretion in the vertebral neuroendocrine system. Because only d-aspartate oxidase (DDO) can degrade d-aspartate, we determined DDO localisation in the pituitary and pineal glands to elucidate the control mechanisms of local d-aspartate concentration. Brain tissues and pituitary and pineal glands of the female pigs contained a similar DDO activity of 0.38–0.66 U/g protein. However, approximately ten-fold higher concentrations of d-aspartate (0.27–0.35 μmol/g protein) were found in both glands. To determine the distribution of immunoreactive DDO, we made a rabbit polyclonal antibody specific to porcine DDO using a recombinant porcine enzyme. DDO immunoreactivity was found in the cytoplasm of a subgroup of cells in the anterior and intermediate lobes, in a part of nerve processes and terminals in the posterior lobe, and in the cytoplasm of a small group of pinealocytes. We used dual-label immunocytochemistry to determine which pituitary hormones colocalise with DDO, and whether DDO and d-aspartate immunoreactivity is reciprocal. In the pituitary gland, almost all proopiomelanocortin-positive cells colocalised DDO, whereas only growth hormone-positive cells colocalised d-aspartate. d-Aspartate immunoreactivity was not detected where DDO immunoreactivity was found. The present study suggests that DDO plays important roles to prevent undesirable off-target action of d-aspartate by strictly controlling local d-aspartate concentration in the pituitary and pineal glands.

M. Shioya, S. Obayashi, H. Tabunoki, K. Arima, Y. Saito, T. Ishida and J. Satoh (2010) Neuropathology and Applied Neurobiology36, 320–330 Aberrant microRNA expression in the brains of neurodegenerative diseases: miR-29a decreased in Alzheimer disease brains targets neurone navigator 3 Aims: MicroRNAs (miRNAs) are small non-coding RNAs that regulate translational repression of target mRNAs. Accumulating evidence indicates that various miRNAs, expressed in a spatially and temporally controlled that manner in the brain plays a key role in neuronal development. However, at present, the pathological implication of aberrant miRNA expression in neurodegenerative events remains largely unknown. To identify miRNAs closely associated with neurodegeneration, we performed miRNA expression profiling of brain tissues of various neurodegenerative diseases. Methods: We initially studied the frontal cortex derived from three amyotrophic lateral sclerosis patients by using a microarray of 723 human miRNAs. This was followed by enlargement of study population with quantitative RT-PCR analysis (n = 21). Results: By microarray analysis, we identified up-regulation of miR-29a, miR-29b and miR-338-3p in amyotrophic lateral sclerosis brains, but due to a great interindividual variation, we could not validate these results by quantitative RT-PCR. However, we found significant down-regulation of miR-29a in Alzheimer disease (AD) brains. The database search on TargetScan, PicTar and miRBase Target identified neurone navigator 3 (NAV3), a regulator of axon guidance, as a principal target of miR-29a, and actually NAV3 mRNA levels were elevated in AD brains. MiR-29a-mediated down-regulation of NAV3 was verified by the luciferase reporter assay. By immunohistochemistry, NAV3 expression was most evidently enhanced in degenerating pyramidal neurones in the cerebral cortex of AD. Conclusions: These observations suggest the hypothesis that underexpression of miR-29a affects neurodegenerative processes by enhancing neuronal NAV3 expression in AD brains.

This study was conducted to investigate the effects of injection of vitamins C (VC) and E (VE) into broodstock on the vitamin levels of broodstock and eggs and subsequent egg and larval quality in the Japanese eel. Broodstock were allotted to one of three groups; control group, injected with physiological saline; early group, injected with vitamins during the early part of maturation (1st–3rd weeks after initiation of maturation induction); later group, injected with vitamins during the later part of maturation (4th–6th weeks after initiation of maturation induction). Broodstock were injected with an emulsion of VC and VE at the same time as an injection of salmon pituitary extract during the induction of maturation. The emulsion was prepared by mixing α-tocopherol, sodium ascorbate, an emulsifier, and water just before injection. The levels of both vitamins in eggs and broodstock increased following vitamin injection. However, the accumulation of vitamins in broodstock and eggs was higher for VE than for VC. Hatching rate, survival and normality of larvae increased with vitamin treatments. The results of this study showed that VC and VE injection into broodstock during artificial maturation increases the vitamin content of eggs and can improve egg quality in Japanese eel.

The corrosion behavior of various kinds of stainless steels was investigated in a high-temperature and high-pressure methanol solution containing chloride ions for the selection of a react...

We investigated the flow properties of mixed solvents of water and N , N -dimethylformamide (DMF) through poly( N -isopropylacrylamide) (PNIPA) gel membranes. The swelling behavior of the gels in the mixed solvents with various compositions was also examined. A simple model to describe the swelling and solvent flow properties for a polymer gel/mixed solvent system was presented. In the swelling measurements, the composition dependence of the degree of swelling ( Θ ) of the PNIPA gels showed a minimum at the DMF fraction ( φ ′ DMF ) of 0.6. In the solvent flow experiments, pressure gradient acting between two solvent–gel interfaces caused the permeation of the mixed solvents through the gel membranes. Little change in solvent composition by the permeation occurred. The integrated flow rate ( F ) over time ( t ) monotonously increased with increasing t . The slope of F versus t curves becomes constant at long times. The inverse of the frictional coefficients for the steady flow plotted against solvent composition is quite similar in shape to the Θ curve. This indicates that the degree of the swelling greatly affects the flow rate of the solvents in the gels.

Antisera directed against hyperpolarization-activated mixed-cation (“ Ih”) and K+ (“Kir”) channels bind to some somata in the ganglion cell layer of rat and rabbit retina. Additionally, the termination of hyperpolarizing current injections can trigger spikes in some cat retinal ganglion cells, suggesting a rebound depolarization arising from activation of Ih. However, patch-clamp studies showed that rat ganglion cells lack inward rectification or present an inwardly rectifying K+ current. We therefore tested whether hyperpolarization activates Ih in dissociated, adult rat retinal ganglion cell somata. We report here that, although we found no inward rectification in some cells, and a Kir-like current in a few cells, hyperpolarization activated Ih in roughly 75% of the cells we recorded from in voltage clamp. We show that this current is blocked by Cs+ or ZD7288 and only slightly reduced by Ba2+, that the current amplitude and reversal potential are sensitive to extracellular Na+ and K+, and that we found no evidence of Kir in cells presenting Ih. In current clamp, injecting hyperpolarizing current induced a slowly relaxing membrane hyperpolarization that rebounded to a few action potentials when the hyperpolarizing current was stopped; both the membrane potential relaxation and rebound spikes were blocked by ZD7288. These results provide the first measurement of Ih in mammalian retinal ganglion cells and indicate that the ion channels of rat retinal ganglion cells may vary in ways not expected from previous voltage and current recordings.

Background Frequent use of emergency rooms by injection drug users (IDUs) has been attributed to a lack of access to primary care and barriers to health services. Using a community-based sample of IDUs, we examined rates of primary care and emergency room use among IDUs and identified correlates of frequent emergency department use. Methods From January to November 2003, we enrolled IDUs into a prospective cohort study involving a baseline questionnaire, comprehensive retrospective and prospective health record linkages. We examined rates of primary care and emergency department utilization, and diagnoses upon arrival in the emergency room. Logistic regression was used to determine factors independently associated with frequent emergency room use. Results Of the 883 IDUs included in this analysis, 687 (78 per cent) accessed a primary care clinic in the previous year, while 528 (60 per cent) participants accessed the emergency room (ER) during the years 2002 and 2003. Abscesses, cellulitis and other skin infections accounted for the greatest proportion of ER use. Factors independently associated with frequent ER use included: frequent crystal methamphetamine injection (AOR = 2.4, 95 per cent CI: 1.0–5.6); non-fatal overdose (AOR = 2.1, 95 per cent CI: 1.4–3.3); HIV-positive status (AOR = 1.5, 95 per cent CI: 1.1–2.1), having been physically assaulted (AOR = 1.5, 95 per cent CI: 1.1–2.1); and primary care utilization (AOR = 1.5, 95 per cent CI: 1.0–2.1). Discussion High rates of ER use were observed among IDUs, despite high rates of primary care use among this same population. ER use was due primarily to preventable injection-related complications that are less amenable to primary care interventions, and therefore educational and prevention efforts that encourage and enable sterile injection practices should be promoted.

We describe here methods for dissociating retinal ganglion cells from adult goldfish and rat without proteolytic enzymes, and show responses of ganglion cells isolated this way to step-wise voltage changes and fluctuating current injections. Taking advantage of the laminar organization of vertebrate retinas, photoreceptors and other cells were lifted away from the distal side of freshly isolated goldfish retinas, after contact with pieces of membrane filter. Likewise, cells were sliced away from the distal side of freshly isolated rat retinas, after these adhered to a membrane filter. The remaining portions of retina were incubated in an enzyme-free, low Ca2+ solution, and triturated. After aliquots of the resulting cell suspension were plated, ganglion cells could be identified by dye retrogradely transported via the optic nerve. These cells showed no obvious morphological degeneration for several days of culture. Perforated-patch whole-cell recordings showed that the goldfish ganglion cells spike tonically in response to depolarizing constant current injections, that these spikes are temporally precise in response to fluctuating current injections, and that the largest voltage-gated Na+ currents of these cells were larger than those of ganglion cells isolated with a neutral protease.

Teeth and periodontal mechanoreceptors play important roles in regulating jaw movements during mastication. However, little is known concerning how jaw movements develop without tooth eruption. To answer this question, we studied masticatory behavior in the osteopetrotic mouse, where tooth eruption does not occur and periodontal mechanoreceptors are missing. A masticatory sequence of the osteopetrotic mouse was divided into two stages: incision and chewing. Incision is characterized by small amplitude and rapid (7 Hz) open-close jaw movements, while slow (5 Hz) and large amplitude open-close jaw movements characterize chewing. The frequency and properties of jaw movements were comparable with those in the normal mouse, though the osteopetrotic mouse had a higher cycle number during incision than did the normal mouse. These results indicate that conversion from sucking to mastication occurs in the anodontic mouse, and the central pattern generator producing the masticatory rhythm develops almost normally without tooth eruption.

Copolymerisation of chlorotrifluoroethylene (CTFE) with alkyl vinyl ester (VES) or alkyl vinyl ether (VE) was studied and the effect of monomer sequence on the weather durability was investigated. The reactivity ratios (r1, r2) determined for the VES/CTFE system and for the VE/CTFE system were obtained by the Fineman-Ross method. VES or VE-centred triad distributions of the copolymers were measured by 13C-NMR and were used to evaluate the completeness of alternating copolymerization with those monomers. Compared with the VES/CTFE system, almost complete alternating sequence was observed in the VE/CTFE system. Then, the weatherability of the polymers coated on the aluminium plates were investigated by the accelerated weathering test, and the results indicated a close correlation between the monomer sequences and the durability of the polymer.

The purpose of the present study was to clarify the relationship between the flow of a bolus and occlusal condition during mastication. First, the characteristics of a bolus under mastication was measured in subjects having different occlusal conditions. Secondly, the flow of a bolus between the upper and lower first molars under mastication was simulated using finite element non-linear dynamic analysis. Measurement of the elasticity of the bolus clarified the phenomenon of its communition. The measurement of the viscosity of the bolus clarified the phenomenon of its mixing with saliva. In addition, a relationship between the elasticity and the viscosity of the bolus at the point of just before swallowing was investigated. The flow of the bolus under mastication was revealed to vary according to the occlusal condition. These results suggest a close relationship between the occlusal condition, the flow of the bolus and its characteristics.

Background: Normative Doppler values and determinants for gender difference of right ventricular diastolic function in healthy subjects have not been fully elucidated. The purpose of this study is to verify the difference of right ventricular diastolic function between men and women at each age. Method: One hundred sixty nine male and 221 female in healthy subject, aged 20 to 90, were examined by both tricuspid inflow velocity and tricuspid annulus velocity by tissue Doppler imaging. All subjects were divided into 7 groups as 20s, 30s, 40s, 50s, 60s, 70s, 80s depend on their age. Results: The tricuspid E velocity was significantly decreased with age (p,0.0001). The tricuspid A velocity was significantly increased with age (p,0.0001). But both E and A velocity were no differences significantly in all generation between men and women (Table). Ratio of tricuspid E and A velocity (E/A) was significantly decreased with age. But no differences were observed in ratio of E/A in all generation between men and women. (20s: 2.0+0.6 vs 2.3+0.6, 30s: 1.9+0.5 vs 2.1+0.5, 40s: 1.6+0.4 vs 1.9+0.5, 50s: 1.3+0.3 vs 1.5+0.4, 60s: 1.2+0.2 vs 1.2+0.3, 70s: 1.1+0.3 vs 1.1+0.3, 80s: 1.2+0.3 vs 1.2+0.3, total: 1.4+0.5 vs 1.5+0.6). Early diastole velocity of tricuspid annulus (Ea) by tissue Doppler method was significantly decreased with age. But Ea was no differences significantly in all generation between men and women (Table). Conclusion: Our results provide the normative Doppler values of right ventricular diastolic function for age decades. There was no difference significantly of right ventricular diastolic function between men and women.

Vertebrates can detect light intensity changes in vastly different photic environments, in part, because postreceptoral neurons undergo “network adaptation.” Previous data implicated dopaminergic, cAMP-dependent inhibition of retinal ganglion cells in this process yet left unclear how this occurs and whether this occurs in darkness versus light. To test for light- and dopamine-dependent changes in ganglion cell cAMP levels in situ , we immunostained dark- and light-adapted retinas with anti-cAMP antisera in the presence and absence of various dopamine receptor ligands. To test for direct effects of dopamine receptor ligands and membrane-permeable protein kinase ligands on ganglion cell excitability, we recorded spikes from isolated ganglion cells in perforated-patch whole-cell mode before and during application of these agents by microperfusion. Our immunostainings show that light, endogenous dopamine, and exogenous dopamine elevate ganglion cell cAMP levels in situ by activating D1-type dopamine receptors. Our spike recordings show that D1-type agonists and 8-bromo cAMP reduce spike frequency and curtail sustained spike firing and that these effects entail protein kinase A activation. These effects resemble those of background light on ganglion cell responses to light flashes. Network adaptation could thus be produced, to some extent, by dopaminergic modulation of ganglion cell spike generation, a mechanism distinct from modulation of transmitter release onto ganglion cells or of transmitter-gated currents in ganglion cells. Combining these observations with results obtained in studies of photoreceptor, bipolar, and horizontal cells indicates that all three layers of neurons in the retina are equipped with mechanisms for adaptation to ambient light intensity.

This study tests the hypothesis that degeneration of a neocortical pathway may facilitate behaviorally-induced growth of neurons in a connected region of the cortex. Degeneration of trancallosal afferents to the motor cortex and changes in forelimb use were independently manipulated in adult rats. The combination of degeneration and behavioral change resulted in the growth of layer V pyramidal neuron dendrites which was not found as a result of either denervation or behavioral manipulation alone. These results indicate that mild degeneration in the adult brain can facilitate neuronal growth when accompanied by appropriate behavioral demand, a finding which has implications for rehabilitative therapy after brain damage.

Multipotent progenitor cells at the retinal margin of adult goldfish give rise to all cell types in the rest of the retina. We took advantage of this spatial arrangement of progenitor and mature cells in slices of peripheral retina, to investigate the appearance and maturation of voltage-activated Na(+) current. We divided the peripheral retina into three broad regions (marginal, intermediate, and mature) on the basis of their morphological development. Whole-cell patch-clamp recordings were performed in ruptured-patch mode, so that cells from which currents were recorded could be identified by Lucifer Yellow fills. No voltage-activated Na(+) current was detected in the slender, peripherally located marginal cells. Voltage-activated Na(+) currents were detected in rounded cells found alongside or near marginal cells, facing the vitreal side of the retina. Some of these "intermediate cells" had a long axon-like process which ran along the vitreal surface. Intermediate cells adjacent to the marginal region tended to have smaller Na(+) currents than intermediate cells closer to the mature region. On average, the maximum Na(+) current amplitude recorded from intermediate cells was roughly 6-fold smaller than that of mature ganglion cells. In addition, the activation threshold of the Na(+) current in intermediate cells was nearly 14 mV more positive than that of mature ganglion cells. The results indicate that voltage-activated Na(+) current, as a possible marker of retinal ganglion cells, begins to develop well before these cells migrate to their adult position within the retina.

SUMMARYIn this study we have tested the concept of using wild-type p53 gene for immunotherapy of cancer. Dendritic cells (DC) were transduced with a human wild-type p53 containing recombinant adenovirus (Ad-p53). About a half of DC transduced with this virus expressed p53 protein by FACS analysis 48 h after infection. Mice immunized twice with Ad-p53 DC developed substantial cytotoxic T lymphocyte (CTL) responses against tumour cells expressing wild-type and different mutant human and murine p53 genes. Very low CTL responses were observed against target cells infected with control adenovirus (Ad-c). Immunization with Ad-p53 provided complete tumour protection in 85% of mice challenged with tumour cells expressing human mutant p53 and in 72.7% of mice challenged with tumour cells with murine mutant p53. Treatment with Ad-p53-transduced DC significantly slowed the growth of established tumours. Thus, DC transduced with wild-type p53 may be a promising new tool for the immunotherapy of cancer.

Methods: Observational cross-sectional analytical study with healthy school students aged 6 to 14 years old from educational institutions in the Greater Florianopolis/SC-Brazil. Biometric data were evaluated (weight, height, body mass index, and body surface area) as predictors and for sample characterization. All participants were submitted for consideration of the respiratory mechanics with the Jaeger impulse oscillometer following ATS standards, with 3 exams on record (data acquisition for at least 20 seconds). For data normality verification the Kolmogorov–Smirnov test and Pearson correlation test were applied identifying the relationship between height, age and weight predicted variables, and oscillometric variation of resistance in 5 and 20 hertz (R5 and R20), reactance (X 5), the respiratory impedance 5 hertz (Z5), resonance frequency (fresh) and resistance area (AX). Models were developed through simple linear regression and multiple analyses. Results: After the recruitment of 864 healthy children, 123 subjects with an average age of 10.02± 2.41 years for boys and 9.98± 2.48 years for girls (p= .94) were considered for final analysis. Correlations were identified between oscillometric dependent and predictor variables, height being considered with greater predictive power in the equations developed for boys in all oscillometric parameters, within an average value of R2 = 55.8%. Age had greater influence on para. Conclusion(s): Reference equations were developed for females and males, considering height as the most influential predictor variable on most IOS parameters in the infant population studied. Implications: Considering the importance of knowledge of pulmonary function and respiratory mechanics in childhood, equations/reference values of the available evaluation methods enable comparisons between healthy children and those who have some respiratory dysfunction.