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Providing plausible strategies for brain aging protection should be a critical concern for countries with large elderly populations including Japan. Age-related cognitive impairments and movement disorders, such as Alzheimer's and Parkinson's diseases, are caused by neurodegeneration that primarily initiates in the hippocampus and the midbrain substantia nigra, respectively. Neurons are postmitotic, and therefore, the accuracy of cellular metabolism should be crucial for maintaining neural functions throughout their life. Thus accuracy of protein synthesis is a critical concern in discussing mechanisms of aging. The essence of the so-called "error catastrophe theory" of aging was on the fidelity of ribosomal translation and/or aminoacylation of tRNA. There is evidence that reduced protein synthesis accuracy results in neurodegeneration. Similarly, reduced proteostasis via autophagy and proteasomes in aging is crucial for protein quality control and well documented as a risk for aging. In both neurodegeneration and protein quality controls, various proteins are involved in their regulation, but recent evidence suggests that repressor element-1 silencing transcription factor (REST) could be a master regulatory protein that is crucial for orchestrating the neural protecting events in human brain aging. REST is induced in the aged brain, and protects neurons against oxidative stress and protein toxicity. Interestingly, REST is identical with neuron-restrictive silencer factor (NRSF), the master regulator of neural development. Thus NRSF/REST play important roles in both neurogenesis and neurodegeneration. In this review, I summarize the interesting scientific crossover, and discuss the potential use of NRSF/REST as a pharmaceutical target for controlling aging, particularly in relation to brain aging.

The endolymphatic sac is a small sac-shaped organ at the end of the membranous labyrinth of the inner ear. The endolymphatic sac absorbs the endolymph, in which the ion balance is crucial for inner ear homeostasis. Of the three sections of the endolymphatic sac, the intermediate portion is the center of endolymph absorption, particularly sodium transport, and is thought to be regulated by aldosterone. Disorders of the endolymphatic sac may cause an excess of endolymph (endolymphatic hydrops), a histological observation in Meniere's disease. A low-salt diet is an effective treatment for Meniere's disease, and is based on the assumption that the absorption of endolymph in the endolymphatic sac abates endolymphatic hydrops through a physiological increase in aldosterone level. However, the molecular basis of endolymph absorption in each portion of the endolymphatic sac is largely unknown because of difficulties in gene expression analysis, resulting from its small size and intricate structure. The present study combined reverse transcription-quantitative polymerase chain reaction and laser capture microdissection techniques to analyze the difference of gene expression of the aldosterone-controlled epithelial Na

Parkinson's disease (PD) is the second most common neurodegenerative disease. Lewy bodies and pale bodies in dopaminergic neurons in the substantia nigra are pathological hallmarks of PD. A number of neurodegenerative diseases demonstrate aggregate formation, but how these aggregates are associated with their pathogenesis remains unknown. It has been reported that repressor element-1 silencing transcription factor/neuron-restrictive silencer factor (REST/NRSF) is induced in the nuclei of aged neurons, preserves neuronal function, and protects against neurodegeneration during aging through the repression of cell death-inducing genes. The loss of REST is associated with Alzheimer's disease pathology. However, its function in dopaminergic neurons remains unknown. Here we demonstrated that REST enters the nucleus of aged dopaminergic neurons. On the other hand, REST is partially sequestrated in Lewy bodies and is mostly absent from the nucleus of neurons in brains with PD and dementia with Lewy bodies (DLB). Dopaminergic neuron-specific autophagy-deficient mice exhibit REST accumulation in aggregates. Defects in the protein quality control system induce REST mRNA expression; its gene product mainly appears in aggregates. Our results suggest that Lewy pathology disturbs normal aging processes in dopaminergic neurons by sequestering REST and the loss of REST may associate with the PD pathology.

Objective The purpose of the study was to investigate the difference in the responses of endolymphatic hydrostatic pressure to isoproterenol, β-adrenergic receptor agonist, between pars superior and pars inferior. Methods The hydrostatic pressure of endolymph and perilymph and endolymphatic potential in the ampulla and the cochlea during the intravenous administration of isoproterenol were recorded using a servo-null system in guinea pigs. Results The hydrostatic pressure of endolymph and perilymph in the ampulla and cochlea was similar in magnitude. Isoproterenol significantly increased hydrostatic pressure of ampullar and cochlear endolymph and perilymph with no change in the ampullar endolymphatic potential and endocochlear potential, respectively. The isoproterenol-induced maximum change of endolymphatic hydrostatic pressure in ampulla was significantly (p

Objective The aim of the study is to evaluate the usefulness of multislice computed tomography (MSCT) using multiplanar reconstruction (MPR) in obtaining preoperative information on the ossicular lesions of middle ear diseases by comparing the ossicular findings of MPR images with the operative findings. Methods Sixty-two ears and 10 ears with preoperative middle ear diseases underwent 4- and 64-detector row CT of the temporal bone in Kagawa University Hospital, respectively. MPR images of three ossicles were created at the planes parallel to the long axis of ossicles. Results The findings of the three ossicles in MPR images were compatible with their operative findings in approximately 91% of 72 ears with various middle ear diseases. There was no significant difference in the coincidence rate of both findings between 4- and 64-detector row CT scanners. The ears with no soft tissue shadows around the ossicles had the coincidence rate of 96–100% in each ossicular part, whereas the coincidence rate was lower in the ears with soft tissue shadows around the ossicles. Conclusion MPR imagings of the ossicles provide accurate preoperative information on the ossicular lesions in middle ear diseases. The 4-detector CT is still a useful device for imaging of the ossicles.

Intracellular Ca2+ levels are changed by influx from extracellular medium and release from intracellular stores. In the central nervous systems, Ca2+ release is involved in various physiological events, such as neuronal excitability and transmitter release. Although stable Ca2+ release in response to stimulus is critical for proper functions of the nervous systems, regulatory mechanisms relating to Ca2+ release are not fully understood in central neurons. Here, we demonstrate that ShcB, an adaptor protein expressed in central neurons, has an essential role in functional maintenance of Ca2+ store in cerebellar Purkinje cells (PCs). ShcB-knockout (KO)mice showed defects in cerebellar-dependent motor function and long-term depression (LTD) at cerebellar synapse. The reduced LTD was accompanied with an impairment of intracellular Ca2+ release. Although the expression of Ca2+ release channels and morphology of Ca2+ store looked intact, content of intracellular Ca2+ store and activity of sarco/endoplasmic reticular Ca2+-ATPase (SERCA) were largely decreased in the ShcB-deficient cerebellum. Furthermore, when ShcB was ectopically expressed in the ShcB-KO PCs, the Ca2+ release and its SERCA-dependent component were restored. These data indicate that ShcB plays a key role in the functional maintenance of ER Ca2+ store in central neurons through regulation of SERCA activity., Scientific Reports, 10(1), art.no.14475; 2020

OBJECTIVES: Tricellulin is a tight junction (TJ)-forming protein that participates in the sealing function of tricellular TJs. Tricellulin-knockout (Tric−/−) mice show progressive hearing loss with degeneration of hair cells in the cochlea without physiological or physical disorders. In the present study, we investigated the tricellulin expression and its deletion effects in the endolymphatic sac (ES) using Tric−/− mice. MATERIALS AND METHODS: The ES epithelia from wild-type (WT) mice were laser-microdissected, and RT-PCR was performed. The ES sections from Tric−/− and WT mice were immunostained with an anti-tricellulin antibody. Hematoxylin and eosin staining was performed for morphological examination. The inner ear of Tric−/− mice was perfused with biotinylation reagents, and the ES sections were observed for tracer permeability assay after applying streptavidin–Alexa Fluor 488 conjugate. RESULTS: The tricellulin expression was confirmed by RT-PCR and by immunohistochemistry in the WT ES. The ES in Tric−/− mice showed normal morphology and revealed no biotin leakage from the lumen. CONCLUSION: The ES in Tric−/− mice showed no changes in morphology or disruption in macromolecular barrier function. The effects of solute leakages in the ES of Tric−/− mice may be very limited and compensatable, or that the ES epithelia may have other sealing system covering the lack of tricellulin.

This study examined whether the forkhead transcription factors of O group 1 (FoxO1) might be involved in telomere biology during calorie restriction (CR). We used FoxO1-knockout heterozygous mice (FoxO1(+/-)) and wild-type mice (WT) as a control. Both WT and FoxO1(+/-) were subjected to ad libitum (AL) feeding or 30% CR compared to AL for 20 weeks from 15 weeks of age. The heart-to-body weight ratio, blood glucose, and serum lipid profiles were not different among all groups of mice at the end of the study. Telomere size was significantly lower in the FoxO1(+/-)-AL than the WT-AL, and telomere attrition was not observed in either WT-CR or FoxO1(+/-)-CR. Telomerase activity was elevated in the heart and liver of WT-CR, but not in those of FoxO1(+/-)-CR. The phosphorylation of Akt was inhibited and Sirt 1 was activated in heart tissues of WT-CR and FoxO1(+/-)-CR. However, the ratio of conjugated to cytosolic light chain 3 increased and the level of p62 decreased in WT-CR, but not in FoxO1(+/-)-CR. A marker of oxidative DNA damage, 8-OhdG, was significantly lower in WT-CR only. The level of MnSOD and eNOS increased, and the level of cleaved caspase-3 decreased in WT-CR, but not FoxO1(+/-)-CR. Echocardiography showed that the left ventricular end-diastolic and systolic dimensions were significantly lower in WT-CR or FoxO1(+/-)-CR than WT-AL or FoxO1(+/-)-AL, respectively. The present studies suggest that FoxO1 plays beneficial roles by inducing genes involved in telomerase activity, as well as anti-oxidant, autophagic, and anti-apoptotic genes under conditions of CR, and suggest that FoxO1 signaling may be an important mediator of metabolic equilibrium during CR.

Introduction Rupture of the extraocular muscle is extremely rare, and a traumatic event is especially uncommon. Although retrieval of an injured MRM is usually performed by an anterior approach by an ophthalmologist, missing muscles in the orbital retrobulbar space are sometimes difficult to identify. This is the first report to demonstrate direct muscle to muscle anastomosis by an endoscopic endonasal orbital approach by a single otolaryngologist. Case report A 67-year-old man presented with left medial rectus muscle (MRM) rupture due to a traffic accident 4 months after injury. The MRM was completely ruptured, and the muscle was repaired by an endoscopic endonasal orbital approach. After surgery, his eye abduction was improved in the primary position, and adduction ability was markedly restored on right gaze without diplopia. Conclusions Our endonasal endoscopic approach provides excellent access to the MRM in the orbital retrobulbar space and avoids a facial scar. We could suture both ends of the muscle together by an endonasal endoscopic approach and could obtain a good result without any complications.

This study describes a technical breakthrough in endolymphatic sac research, made possible by the use of the recently generated Prox1-GFP transgenic mouse model. Whole-mount imaging techniques through the decalcified temporal bone and three-dimensional observations of Prox1-GFP mouse tissue revealed the positive labeling of the endolymphatic sac in adult stage, and allowed, for the first time, the GFP-based identification of endolymphatic sac epithelial cells. Prox1 expression was observed in all parts of the endolymphatic sac epithelia. In intermediate portion of the endolymphatic sac, mitochondria-rich cells did not express Prox1, although ribosome-rich cells showed strong GFP labeling. The anatomical relationship between the endolymphatic sac and the surrounding vasculature was directly observed. In the endolymphatic sac, expression of Prox1 may suggest progenitor cell-like pluripotency or developmental similarity to systemic lymphatic vessels in other organs. This whole-mount imaging technique of the endolymphatic sac can be combined with other conventional histological, sectioning, and labeling techniques and will be very useful for future endolymphatic sac research.

Objective Silver-containing carboxymethylcellulose fiber dressing (Aquacel ® -Ag) has been used to treat burns and ulcers with a large amount of exudate. The aim of this investigation was to confirm whether Aquacel ® -Ag has beneficial effects when it is used as nasal packing. Methods We included 44 patients who underwent bilateral endoscopic sinus surgery due to chronic rhino-sinusitis. Beschitin-F ® or Aquacel ® -Ag was packed postoperatively into the bilateral middle meatus. Patient's comfort was recorded using a VAS, as well as wound healing, postoperative bleeding and local infection. Postoperative-specific organisms were also evaluated from the removed packing materials located in the middle meatus when they were removed on the 4th day after surgery. Results The scores for nasal obstruction and pain were not statistically different in each group. Postoperative bacteriologic studies indicated marked differences. Coagulase-negative staphylococci were predominant and potential pathogens were recovered in a few cases in the Aquacel ® -Ag group. In contrast, potential pathogens, including Streptococcus pneumonia , Haemophilus influenza , and Gram-negative rods, were predominant in the Beschitin-F ® group. Conclusion The results indicate that Aquacel ® -Ag might contribute to hemostasis, wound healing, and patient comfort after endonasal surgery, similar to Beschitin-F ® . Additionally, it may have advantages concerning the prevention of postoperative infection.

Nasal resistance and the OSA-18 score were useful for evaluating surgical treatments. The sleep disturbance score may also be useful for predicting the severity of obstructive sleep apnea (OSA) in children.We evaluated the effect of surgery on children with OSA using polysomnography (PSG) parameters, nasal resistance, and the OSA-18 questionnaire, and also investigated the cut-off OSA-18 score to screen for pediatric OSA.This was a retrospective study in which PSG parameters and nasal resistance were measured using a rhinomanometer and the OSA-18 score was obtained from the OSA-18 questionnaire before and after surgery in 45 children with OSA.The mean age of the 45 patients was 5.7 ± 2.0 years. The mean value of the obstructive apnea hypopnea index (O-AHI) improved from 16.2 ± 14.3/h before surgery to 1.1 ± 1.7/h after surgery, the mean nasal resistance improved from 0.44 ± 0.19 to 0.32 ± 0.10 Pa/cm(3)/s, and the mean OSA-18 score improved from 61.1 ± 13.7 to 30.4 ± 5.8, and all these improvements were significant. The O-AHI value was lower than 1/h after surgery in 64.4% of patients (29/45). The O-AHI value was significantly correlated with the sleep disturbance score (r = 0.352, p = 0.018). When the cut-off OSA-18 score for screening was set at 40, sensitivity was 100%.

Intranasal corticosteroids are effective for allergic rhinitis and broadly used in daily clinical practice. Systemic oral corticosteroids are also known to be effective for treatment of allergic rhinitis. These topical and systemic corticosteroids are both effective formulations for allergic rhinitis, and both drugs have some side effects. When treatment formulations for allergic rhinitis are selected based on side effects, topical corticosteroids are more commonly selected than systemic steroids. Systemic corticosteroids, on the other hand, have traditionally been believed to have higher and more instantaneous therapeutic effects than those of topical corticosteroids. However, there have been few reports of direct comparisons between topical corticosteroid and systemic corticosteroid efficacy. The purpose of this study was to evaluate the subjective outcomes of nasal symptom management using topical intranasal corticosteroid therapy or systemic oral corticosteroid therapy in patients with seasonal allergic rhinitis. We compared the efficacy of mometasone furoate nasal spray (MFNS) to betamethasone oral tablets (BOT) for the treatment of patients with seasonal allergic rhinitis.In an open label study, patients with seasonal allergic rhinitis who had intermediate-to-severe symptoms and who visited the hospital without prior treatment were allocated to 1 of 3 treatment groups (noncorticosteroid group, topical corticosteroid group, and oral corticosteroid group). Evaluation was conducted using allergy diaries that consisted of patient questionnaires. The Japanese Rhinoconjunctivitis Quality of Life Questionnaire (JRQLQ) was used in this study.Compared to only loratazine nonsteroid therapy, both MFNS 200μg once daily and BOT 0.25mg twice daily significantly reduced the total and individual symptom scores for sneezing, nasal obstruction, watery nasal discharge, and nasal itching (P0.05). Scores for itching of the eyes were reduced slightly more in the MFNS group than in the noncorticosteriod treatment group, but the difference was not significant.MFNS and BOT have virtually equivalent effects on nasal symptoms in patients with seasonal allergies. Our study was the first direct comparison between an intranasal corticosteroid spray and a systemic oral corticosteroid for seasonal allergic rhinitis. No significant differences were found in the therapeutic effects of the topical and systemic corticosteroids tested, suggesting that topical corticosteroids are expected to sufficiently improve nasal symptoms without administration of oral corticosteroids. Treatment with intranasal corticosteroid spray is more strongly recommended than treatment with systemic corticoid steroids, due to the side effects associated with each treatment.

Objective The aim of this study was to assess the value of multislice computed tomography (MSCT) using multiplanar reconstruction (MPR) to detect the small fenestral lesions in patients with otosclerosis. Methods MSCT with MPR imaging was used to evaluate 27 ears of 17 patients with otosclerosis (3 male and 14 females) ranging in age from 33 to 69 years with a mean of 49.8 year. MSCT imaging was performed using Aquilion ® . Axial spiral scans with a 0.5-mm slice thickness were obtained. The acquired high-resolution data were transferred to a workstation (ALATO VIEW). MPR images were created in the planes parallel to the stapedial crus and then analyzed on the monitor screen by two radiologists (Y.T. and N.K.). Results MPR images showed abnormal findings in 26 of 27 ears with otosclerosis (96%), whereas axial images showed abnormal findings in only 15 of 27 ears (56%). The similar classification between both images was shown only in 9 of 27 ears (33%). In 16 ears (67%) axial images under-evaluated the lesions compared with MPR images. MPR images detected smaller foci than axial images. Air-bone gap at 0.5–4 kHz tended to increase dependently on the extension of fenestral lesions evaluated by MPR images. Conclusion MPR images detected fenestral lesions in otosclerosis more frequently and more precisely than axial images. The extent of fenestral lesions observed on MPR images tends to be related to the degree of conductive hearing loss.

BDNF-TrkB signaling is implicated in experimental seizures and epilepsy. However, the downstream signaling involved in the epileptiform activity caused by TrkB receptor activation is still unknown. The aim of the present study was to determine whether TrkB-mediated N-Shc signal transduction was involved in kainic acid (KA)-induced epileptiform activity. We investigated KA-induced behavioral seizures, epileptiform activities and neuronal cell loss in hippocampus between N-Shc deficient and control mice. There was a significant reduction in seizure severity and the frequency of epileptiform discharges in N-Shc deficient mice, as compared with wild-type and C57BL/6 mice. KA-induced neuronal cell loss in the CA3 of hippocampus was also inhibited in N-Shc deficient mice. This study demonstrates that the activation of N-Shc signaling pathway contributes to an acute KA-induced epileptiform activity and neuronal cell loss in the hippocampus. We propose that the N-Shc-mediated signaling pathway could provide a potential target for the novel therapeutic approaches of epilepsy.

This study compared the utility of 3'-deoxy-3'-(18)F-fluorothymidine PET ((18)F-FLT PET) with that of (18)F-FDG PET for assessment of the early locoregional clinical outcomes of chemoradiotherapy for head and neck squamous cell carcinomas.From May 2006 to September 2010, 28 patients with head and neck squamous cell carcinomas underwent (18)F-FLT and (18)F-FDG PET before radiation therapy (RT), 4 wk after the initiation of RT, and 5 wk after completion of RT. PET images were evaluated qualitatively for regions of focally increased metabolism and were analyzed in relation to residual accumulation and local disease control.During RT, (18)F-FLT uptake decreased more significantly than (18)F-FDG uptake. (18)F-FLT accumulations disappeared in 34 of 54 lesions (63%), and negative predictive value was 97%. (18)F-FDG PET during RT also had a high negative predictive value (100%), but only 9 lesions (16%) showed complete absence of accumulation. The specificity and overall accuracy of (18)F-FLT PET were significantly higher than those of (18)F-FDG PET both during and after RT. In particular, high significance was attributable to the results of the evaluations of primary lesions. There were significant differences in 3-y local control between the residual-accumulation and no-accumulation groups on both posttreatment (18)F-FLT PET (P0.0001) and posttreatment (18)F-FDG PET (P = 0.0081).(18)F-FLT PET during RT and early follow-up facilitates the selection of optimal further therapy and the prediction of outcomes.

Objectives/Hypothesis: Pott's puffy tumor (PPT) is defined as one or more subperiosteal abscesses of the frontal bone based on osteomyelitis. PPT is observed predominantly in the adolescent age group and rarely in adults. Some parameters affecting prognosis and an appropriate surgical approach for antecedent frontal sinusitis have not been elucidated due to the rarity of patients with adult PPT. Study Design: Retrospective patient record and literature study. Methods: Five patients from our cohort and 27 patients identified in a literature search formed the study group. Results: The incidence rate of intracranial complications was lower than in previous reports at 29.0%. There was no correlation between the incidence rate of intracranial complications and each patient's parameters. It was indicated that the department first consulted by the patients was possibly related to the initial diagnosis and the incidence rate of intracranial complications. Conclusions: Although the incidence rate of major complications is lower than in children and later than in earlier published adult cases, patients are still at high risk of serious intracranial complications. Early diagnosis and adequate treatment may contribute to reducing the incidence rate. Laryngoscope, 2012

Objective Claudins are a family of membrane proteins which localize to tight junctions (TJs). Recent studies have shown that claudins can form pores for ions in the TJs and regulate the permeability of epithelial paracellular ion transport. The endolymphatic sac (ES) is a part of the inner ear, absorbing the endolymphatic fluid. ES dysfunction may result in endolymphatic hydrops. In this study, we focused on the paracellular transport and examined claudin mRNA expression in the ES epithelia. Materials and methods Total RNA was isolated from whole ES epithelia of rats by laser capture microdissection. RT-PCR was used to evaluate the expression of claudins. The expression of each claudin mRNA in the epithelial cells of rat ES was confirmed by in situ hybridization. Results RT-PCR indicated the expression of cldn2, cldn4, cldn6, cldn7, cldn9, cldn11, cldn12, and cldn14 . The expression of these claudin mRNAs in the epithelial cells of rat ES was confirmed by in situ hybridization. Conclusion We demonstrated mRNA expression of multiple claudins in the rat ES epithelia. These results in the ES epithelia were consistent with a role of claudins in paracellular ion transport.

Basal cell adenocarcinoma (BCAC) is a rare malignant neoplasm in the salivary glands and BCAC of the minor salivary glands is exceedingly rare. Only nine cases of palatal BCACs of the minor salivary gland have been reported. BCAC is a low-grade malignant tumour which shares many histologic characteristics with basal cell adenoma. Histological differentiation between the two is difficult and they are often discriminated only by invasion of local structures or by perineural or vascular invasive figures. The authors describe the case of a 69-year-old man with a massive BCAC of a palatal minor salivary gland that progressed into the nasal cavity and pterygopalatine fossa and was treated by a subtotal maxillectomy. This is a highly locally advanced case which required a wider surgical excision range than other previously reported BCAC cases of the palatal minor salivary glands. In this case, the proper diagnosis could not be made by local biopsy alone. It should be kept in mind that it may be difficult to distinguish BCAC from basal cell adenoma by microscopic examination of biopsy specimens alone.

Oxidative stress is a primary factor inducing brain dysfunction in aged animals. However, how oxidation affects brain function is not fully understood. Here we show that oxidation inhibits signaling pathways essential for synaptic plasticities in the cerebellum. We first revealed that nitric oxide (NO)-dependent plasticities at the parallel fiber-Purkinje cell synapse (PF synapse) were impaired in the cerebellar slices from aged mice, suggesting a possible inhibitory action of protein oxidation by endogenous reactive oxygen species. PF-synaptic plasticities were also blocked in the cerebellar slices from young mice preincubated with oxidizing agents or thiol blocker. Because the treatment of the slices with the oxidizing agent did not affect basic electrophysiological properties of excitatory postsynaptic current of PF (PF-EPSC) and did not occlude the synaptic plasticities, oxidation was revealed to specifically inhibit signaling pathways essential for PF-synaptic plasticities. Finally, biochemical analysis confirmed the idea that inhibitory action of protein oxidation on the PF-synaptic plasticities was mediated by impairment of nitric oxide-induced protein S-nitrosylation. Therefore, oxidation was revealed to inhibit the S-nitrosylation-dependent signaling pathway essential for synaptic plasticity in a "competitive" manner., Neurobiology of Aging, 33(3), pp.535-545; 2012

The phosphotyrosine signaling followed by various receptor activations conforms a unique signaling platform during metazoan evolution, and is crucial for animal development, maturation, and aging. Shc is the most versatile bipartite phosphotyrosine signal adaptor harboring phosphotrosine-biding (PTB) and Src-homology2 (SH2) domains. Among the Shc adaptor family members, p66-Shc is of potential interest in aging studies, since its deletion in mice resulted in a longer lifespan and/or higher quality of life in later stages of life. However, a few studies have examined the gene expression profiles of p66-Shc in aging tissues. Here, we quantified the expression levels of transcripts of Shc-related isoforms in the liver and brain of young adult, middle-aged, and aged rats, and found that p66-Shc gene expression is specifically up-regulated in the aged liver and brain. In the aged liver tissue, p66-Shc expression was also evident at the protein level, and accumulated in the soluble fraction of the aged tissue. These results indicate that p66-Shc is not only related to animal longevity but also affected during aging, and thus the repression of p66-Shc could become a potential target for an anti-aging strategy., Archives of Gerontology and Geriatrics, 55(3), pp. 744-748; 2012

Mobilization of intracellular Ca2+ stores regulates a multitude of cellular functions, but the role of intracellular Ca2+ release via the ryanodine receptor (RyR) in the brain remains incompletely understood. We found that nitric oxide (NO) directly activates RyRs, which induce Ca2+ release from intracellular stores of central neurons, and thereby promote prolonged Ca2+ signalling in the brain. Reversible S-nitrosylation of type 1 RyR (RyR1) triggers this Ca2+ release. NO-induced Ca2+ release (NICR) is evoked by type 1 NO synthase-dependent NO production during neural firing, and is essential for cerebellar synaptic plasticity. NO production has also been implicated in pathological conditions including ischaemic brain injury, and our results suggest that NICR is involved in NO-induced neuronal cell death. These findings suggest that NICR via RyR1 plays a regulatory role in the physiological and pathophysiological functions of the brain.

We demonstrated previously that the blood pressure of patients with IgA nephropathy becomes salt sensitive as renal damage progresses. We also showed that increased urinary angiotensinogen levels in such patients closely correlate with augmented renal tissue angiotensinogen gene expression and angiotensin II levels. Here, we investigated the relationship between urinary angiotensinogen and salt sensitivity of blood pressure in patients with IgA nephropathy. Forty-one patients with IgA nephropathy consumed an ordinary salt diet (12 g/d of NaCl) for 1 week and a low-salt diet (5 g/d of NaCl) for 1 week in random order. The salt-sensitivity index was calculated as the reciprocal of the slope of the pressure-natriuresis curve drawn by linking 2 data points obtained during consumption of each diet. The urinary angiotensinogen:creatinine ratio was significantly higher in patients who consumed the ordinary salt diet compared with the low-salt diet (17.5 μg/g [range: 7.3 to 35.6 μg/g] versus 7.9 μg/g [range: 3.1 to 14.2 μg/g] of creatinine, respectively; P r =0.43; P =0.008) and changes in logarithmic urinary angiotensinogen:creatinine ratio ( r =0.37; P =0.017) but not with changes in urinary protein excretion ( r =0.18; P =0.49). In contrast, changes in sodium intake did not alter the urinary angiotensinogen:creatinine ratio in patients with Ménière disease and normal renal function (n=9). These data suggest that the inappropriate augmentation of intrarenal angiotensinogen induced by salt and associated renal damage contribute to the development of salt-sensitive hypertension in patients with IgA nephropathy.

OBJECTIVE: To determine the utility of 2-deoxy-2-[F]fluoro-D-glucose positron emission tomography (FDG-PET) for assessment of early locoregional clinical outcome of chemoradiotherapy for head and neck squamous cell carcinomas (HNSCCs). METHODS: From July 2002 to July 2008, a total of 35 patients with HNSCCs underwent FDG-PET before and between 4 and 7 weeks (median: 5 weeks) after the end of concurrent chemoradiotherapy. FDG uptake of primary sites and metastatic lymph nodes was measured as standardized uptake values (SUVs) and was analyzed in relation to local control. RESULTS: Visual inspection showed a sensitivity of 88.2% and a negative predictive value of 93.1%, but the positive predictive value was only 37.5%. Setting a posttreatment SUV of 3.0 and percentage change of 60 as cut-off values, positive predictive value increased to 81.3% and overall accuracy increased to 89.9% using the combined analysis of these two parameters. The local control rate was not associated with T, the N category, or the tumor node metastasis stage. In contrast, combined analysis of the posttreatment SUV and the percentage change in SUV was a useful indicator of locoregional control. In particular, combined evaluation of SUVs was very effective for assessment of therapeutic effects with reference to metastatic lymph nodes. CONCLUSION: Early follow-up with FDG-PET is thus considered helpful further in choosing optimal therapy and for making an accurate prognosis.

Neural-restrictive silencer (NRS) has been well characterized in SCG10 and many other neuron-specific genes; it is, however, unknown whether the promoters of the SCLIP and RB3 genes (two other SCG10 family members) share basal transcriptional mechanisms with SCG10 or not. To explore how NRS-mediated neural-specific gene transcription has evolved, we determined the genomic and promoter structures of the SCLIP gene, and found that the gene retained an NRS-like element that functioned as a negative regulator in non-neuronal cells. However, unlike the NRS in the SCG10 gene, this NRS(SCLIP) was located downstream of the transcription start site, and showed a position-dependent repressing activity. Further gel-shift and NRS factor (NRSF) co-transfection experiments revealed that NRS(SCLIP) was bound and regulated by NRSF. Such an element was not found in the gene promoter of RB3, suggesting that the NRS-NRSF regulatory system evolved once SCLIP had diverged from RB3 or stathmin.

The flavoprotein rotenone-insensitive internal NADH-ubiquinone (UQ) oxidoreductase (Ndi1) is a member of the respiratory chain in Saccharomyces cerevisiae. We reported previously that bound UQ in Ndi1 plays a key role in preventing the generation of reactive oxygen species. Here, to elucidate this mechanism, we investigated biochemical properties of Ndi1 and its mutants in which highly conserved amino acid residues (presumably involved in NADH and/or UQ binding sites) were replaced. We found that wild-type Ndi1 formed a stable charge transfer (CT) complex (around 740 nm) with NADH, but not with NADPH, under anaerobic conditions. The intensity of the CT absorption band was significantly increased by the presence of bound UQ or externally added n-decylbenzoquinone. Interestingly, however, when Ndi1 was exposed to air, the CT band transiently reached the same maximum level regardless of the presence of UQ. This suggests that Ndi1 forms a ternary complex with NADH and UQ, but the role of UQ in withdrawing an electron can be substitutable with oxygen. Proteinase K digestion analysis showed that NADH (but not NADPH) binding induces conformational changes in Ndi1. The kinetic study of wild-type and mutant Ndi1 indicated that there is no overlap between NADH and UQ binding sites. Moreover, we found that the bound UQ can reversibly dissociate from Ndi1 and is thus replaceable with other quinones in the membrane. Taken together, unlike other NAD(P)H-UQ oxidoreductases, the Ndi1 reaction proceeds through a ternary complex (not a ping-pong) mechanism. The bound UQ keeps oxygen away from the reduced flavin.

Acute respiratory distress syndrome (ARDS) は高サイトカイン血症に伴う活性化好中球による血管内皮障害と，それに引き続く肺胞上皮細胞障害による肺水腫が病態の中心であり，その死亡率は30％以上とされている。われわれは化学療法後の好中球減少期に肺炎に罹患し，G-CSF製剤投与後，白血球数の上昇とともに呼吸状態が悪化しARDSとなった症例を経験したので報告する。症例は70歳男性。中咽頭癌cT4aN2cM1 (stage IV C) に対し，ドセタキセル83 mg (60 mg/m2) による化学療法を施行した。Day8に好中球数が61/μlとなりDay8からDay10までレノグラスチム100 μgを投与し，Day11に好中球数は8275/μlまで回復した。Day10よりCRP値の上昇を伴う呼吸困難が出現し，肺炎として治療を開始したが，Day13にはさらに呼吸機能が悪化し，肺炎による呼吸機能低下，播種性血管内凝固症候群として加療を行った。しかし，Day14にはさらに酸素化が不十分となり人工呼吸管理となった。ARDSの診断にてDay14よりDay21までシベレスタットナトリウム200 mgを投与し，またコハク酸ヒドロコルチゾンナトリウムを200 mgから漸減しながら投与した。その後徐々に呼吸機能は改善した。G-CSF投与後の白血球回復期と肺炎罹患が重複した場合，ARDSなどの肺障害に対する注意が必要であると思われた。

Accumulation of ubiquitinated protein aggregates is a common pathology associated with a number of neurodegenerative diseases and selective autophagy plays a critical role in their elimination. Although aging-related decreases in protein degradation properties may enhance protein aggregation, it remains unclear whether proteasome dysfunction is indispensable for ubiquitinated-protein aggregation in neurodegenerative diseases. Here, we show that N-oleoyl-dopamine and N-arachidonyl-dopamine, which are endogenous brain substances and belong to the N-acyldopamine (AcylDA) family, generate cellular inclusions through aggresome formation without proteasome inhibition. Although AcylDA itself does not inhibit proteasome activity in vitro, it activates the rearrangement of vimentin distribution to form a vimentin cage surrounding aggresomes and sequesters ubiquitinated proteins in aggresomes. The gene transcription of p62/SQSTM1 was significantly increased by AcylDAs, whereas the transcription of other ubiquitin-dependent autophagy receptors was unaffected. Genetic depletion of p62 resulted in the loss of ubiquitinated-protein sequestration in aggresomes, indicating that p62 is a critical component of aggresomes. Furthermore, AcylDAs accelerate the aggregation of mutant huntingtin exon 1 proteins. These results suggest that aggresome formation does not require proteasome dysfunction and AcylDA-induced aggresome formation may participate in forming cytoplasmic protein inclusions., Scientific Reports, 8, 9585; 2018

The FoxO transcription factors may be involved in the antiaging effect of calorie restriction (CR) in mammals. To test the hypothesis, we used FoxO1 knockout heterozygotic (HT) mice, in which the FoxO1 mRNA level was reduced by 50%, or less, of that in wild-type (WT) mouse tissues. The WT and HT mice were fed ad libitum (AL) or 30% CR diets from 12 weeks of age. Aging- and CR-related changes in body weight, food intake, blood glucose, and insulin concentrations were similar between the WT and HT mice in the lifespan study. The response to oxidative stress, induced by intraperitoneal injection of 3-nitropropionic acid (3-NPA), was evaluated in the liver and hippocampus at 6 months of age. Several of the selected FoxO1-target genes for cell cycle arrest, DNA repair, apoptosis, and stress resistance were up-regulated in the WT-CR tissues after 3-NPA injection, while the effect was mostly diminished in the HT-CR tissues. Of these gene products, we focused on the nuclear p21 protein level in the liver and confirmed its up-regulation only in the WT-CR mice in response to oxidative stress. The lifespan did not differ significantly between the WT and HT mice in AL or CR conditions. However, the antineoplastic effect of CR, as indicated by reduced incidence of tumors at death in the WT-CR mice, was mostly abrogated in the HT-CR mice. The present results suggest a role for FoxO1 in the antineoplastic effect of CR through the induction of genes responsible for protection against oxidative and genotoxic stress., Aging cell, 9(3), pp.372-382; 2010

Neurofibrillary tangles composed of hyperphosphorylated tau protein are primarily neuropathological features of a number of neurodegenerative diseases collectively termed tauopathy. To understand the mechanisms underlying the cause of tauopathy, precise cellular and animal models are required. Recent data suggest that the transient introduction of exogenous tau can accelerate the development of tauopathy in the brains of non-transgenic and transgenic mice expressing wild-type human tau. However, the transmission mechanism leading to tauopathy is not fully understood. In this study, we developed cultured-cell models of tauopathy representing a human tauopathy. Neuro2a (N2a) cells containing propagative tau filaments were generated by introducing purified tau fibrils. These cell lines expressed full-length (2N4R) human tau and the green fluorescent protein (GFP)-fused repeat domain of tau with P301L mutation. Immunocytochemistry and super-resolution microscopic imaging revealed that tau inclusions exhibited filamentous morphology and were composed of both full-length and repeat domain fragment tau. Live-cell imaging analysis revealed that filamentous tau inclusions are transmitted to daughter cells, resulting in yeast-prion-like propagation. By a standard method of tau preparation, both full-length tau and repeat domain fragments were recovered in sarkosyl insoluble fraction. Hyperphosphorylation of full-length tau was confirmed by the immunoreactivity of phospho-Tau antibodies and mobility shifts by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). These properties were similar to the biochemical features of P301L mutated human tau in a mouse model of tauopathy. In addition, filamentous tau aggregates in cells barely co-localized with ubiquitins, suggesting that most tau aggregates were excluded from protein degradation systems, and thus propagated to daughter cells. The present cellular model of tauopathy will provide an advantage for dissecting the mechanisms of tau aggregation and degradation and be a powerful tool for drug screening to prevent tauopathy., International Journal of Molecular Sciences, 19(5), 1497; 2018

We compared 3'[F-18]fluoro-3'-deoxythymidine (FLT) positron emission tomography (PET) and 2-deoxy-2-[F-18]fluoro-D-glucose (FDG) for PET visualization of head and neck squamous cell cancers (HNSCCs) and evaluated which might better reflect proliferative activity as indicated by the Ki-67 index. A total of 43 patients with HNSCCs were examined with FLT-PET and FDG-PET. The PET images were evaluated qualitatively for regions of focally increased metabolism and for semiquantitative analysis the maximum standardized uptake value (SUV) was calculated. For depiction of primary tumours, the sensitivity of both approaches was 100%. The mean (±SD) SUV for FLT (5.65 ± 2.96) was significantly lower than that for FDG (10.9 ± 4.91; p

The rat pheochromocytoma cell line PC12 has been widely used as a model to study neuronal differentiation. PC12 cells give rise to neurites in response to basic fibroblast growth factor (bFGF). However, it is unclear whether bFGF promotes neurite outgrowth by inducing RhoA inactivation, and a mechanism for RhoA inactivation in PC12 cells in response to bFGF has not been reported. Lysophosphatidic acid (LPA) treatment and the expression of constitutively active (CA)-RhoA (RhoA V14) impaired neurite formation in response to bFGF, while Tat-C3 exoenzyme and the expression of dominant negative (DN)-RhoA (RhoA N19) stimulated neurite outgrowth. GTP-bound RhoA levels were reduced in response to bFGF, which suggests that the inactivation of RhoA is essential to neurite outgrowth in response to bFGF. To investigate the mechanism of RhoA inactivation, this study examined the roles of p190RhoGAP and Rap-dependent RhoGAP (ARAP3). DN-p190RhoGAP prevented neurite outgrowth, while WT-p190RhoGAP and Src synergistically stimulated neurite outgrowth; these findings suggest that bFGF promotes the inactivation of RhoA and subsequent neurite outgrowth through p190RhoGAP and Src. Furthermore, DN-Rap1 and DN-ARAP3 reduced neurite formation in PC12 cells. These results suggest that RhoA is likely to be inactivated by p190RhoGAP and ARAP3 during neurite outgrowth in response to bFGF.

The FoxO transcription factors may be involved in the antiaging effect of calorie restriction (CR) in mammals. To test the hypothesis, we used FoxO1 knockout heterozygotic (HT) mice, in which the FoxO1 mRNA level was reduced by 50%, or less, of that in wild-type (WT) mouse tissues. The WT and HT mice were fed ad libitum (AL) or 30% CR diets from 12 weeks of age. Aging- and CR-related changes in body weight, food intake, blood glucose, and insulin concentrations were similar between the WT and HT mice in the lifespan study. The response to oxidative stress, induced by intraperitoneal injection of 3-nitropropionic acid (3-NPA), was evaluated in the liver and hippocampus at 6 months of age. Several of the selected FoxO1-target genes for cell cycle arrest, DNA repair, apoptosis, and stress resistance were up-regulated in the WT-CR tissues after 3-NPA injection, while the effect was mostly diminished in the HT-CR tissues. Of these gene products, we focused on the nuclear p21 protein level in the liver and confirmed its up-regulation only in the WT-CR mice in response to oxidative stress. The lifespan did not differ significantly between the WT and HT mice in AL or CR conditions. However, the antineoplastic effect of CR, as indicated by reduced incidence of tumors at death in the WT-CR mice, was mostly abrogated in the HT-CR mice. The present results suggest a role for FoxO1 in the antineoplastic effect of CR through the induction of genes responsible for protection against oxidative and genotoxic stress.

Aspergillosis, a fungal infection generally acquired by inhaling airborne spores (conidia) of the ubiquitous mold Aspergillus, may cause symptomatic infection such as allergic aspergillosis in young atopic subjects, aspergillomas also called fungus balls, and invasive aspergillosis typically seen in debilitated or immunocompromised subjects. A 71-year-old man who had undergone chemoradiotherapy for malignant lymphoma 12 years earlier was seen for a visual disorder and pain. It was found in computed tomography (CT) and magnetic resonance imaging (MRI) to have a right orbital apex lesion but no paranasal sinus abnormalities. Based on a diagnosis of optic neuritis without a histopathological diagnosis, we started systemic steroid therapy, which only worsened his symptoms. Endoscopic endonasal orbital apex biopsy conducted afterwards for a definitive diagnosis found that the visual disorder was caused by invasive fungal infection, underscoring the importance of pathological examination.