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The authors conducted droplet evaporation experiments of light cycle oil (LCO) at various ambient temperatures and pressures. Five kinds of LCO and three kinds of arranged fuels were used. We investigated the evaporation characteristics of LCO and the relationships between the evaporation characteristics and the cetane index. In addition to that, a surrogate fuel composed of four chemical species, which can simulate the droplet evaporation characteristics of LCO, was suggested. Experimental results show that the differences in droplet lifetime between fuel species become larger with decreasing ambient temperature. This is because the low volatile component made the evaporation rate outstandingly slow at a low ambient temperature. It was found that the relationship between droplet lifetime and the late-stage distillation temperature becomes stronger at low ambient temperature and high ambient pressure. By an analysis employing the properties of chemical species in LCO surrogate fuel, it is clarified that the mass evaporation rate becomes smaller than the internal diffusion, which is the condition similar to that in the distillation test. Finally, the relationship between the droplet lifetime and the cetane index was investigated. It can be concluded that the droplet lifetime is independent of the cetane index under all conditions tested in this study. The experimental data obtained by this research can be utilized for the validation of multicomponent fuel droplet evaporation models in the future.

Daikenchuto (DKT) is one of the most widely used “Kampo” in Japan as a representative of herbal medicine. Because DKT is made from a natural product like food, it requires the management of pesticides; therefore, an analysis of residual pesticides in Kampo is required. The World Health Organization (WHO) indicates that pesticide residue analysis by the U.S. Pharmacopeia (USP) is required. USP defines 107 compounds containing organochlorine pesticides and organophosphorus pesticides and their metabolites, which have a high residual risk. Accordingly, to guarantee the safety of herbal medicines according to global standards is a very important issue. In this study, we developed an analytical method for 91 compounds, which are listed in USP, using DKT as the subject. The method could extract pesticides from DKT with acetone, elute pesticides with acetonitrile using a SepPak C18 column (5 g) and with ethyl acetate using a DSC-NH2 column (2 g), and perform simultaneous analyses by gas chromatography–tandem mass spectrometry (GC–MS/MS). This method, which could quantify 88 compounds, was validated according to USP. A pesticide residue analysis method that meets USP requirements enables the analysis of pesticide residues with a high residue risk and contributes to improving the safety of “Kampo” and other herbal medicines.

Background Glyphosate and its salt formulations are nonselective herbicides that have been extensively used worldwide, both for residential and agricultural purposes. The possible carcinogenicity and teratogenicity of glyphosate remain to be elucidated. We developed a sensitive and high-throughput analytical method for urinary glyphosate using liquid chromatography-tandem mass spectrometry with the aim of contributing to glyphosate exposure assessment in epidemiological studies. Methods After urine dilution (creatinine matching dilution to 0.05 g creatinine/L), glyphosate was extracted using two types of solid phase extraction columns (SCX and NH2) with automated sample preparation instruments. The eluate was dried and dissolved in the mobile phase, followed by liquid chromatography-tandem mass spectrometry analysis. The optimized method was applied to urine samples obtained from 54 Japanese adults and children. Results The results from the validation study demonstrated good recoveries (91.0-99.6%), within- and between-run precisions (< 15%), low detection limits (0.1 μg/L), and lower limit of quantification (0.3 μg/L). The detection frequency and median concentration of the urinary glyphosate in Japanese subjects were 59% and 0.25 μg/L (0.34 μg/g creatinine). Conclusions Our reliable determination method was successful in measuring urinary glyphosate concentration. Moreover, this is the first biomonitoring report of urinary glyphosate levels in the Japanese general population.

The comorbidity of chronic pain and mental dysfunctions such as depression and anxiety disorders has long been recognized, but the underlying mechanisms remain poorly understood. Here, using a mouse model of neuropathic pain, we demonstrated neuronal plasticity in the bed nucleus of the stria terminalis (BNST), which plays a critical role in chronic pain–induced maladaptive anxiety. Electrophysiology demonstrated that chronic pain increased inhibitory inputs to lateral hypothalamus (LH)–projecting BNST neurons. Chemogenetic manipulation revealed that sustained suppression of LH-projecting BNST neurons played a crucial role in chronic pain–induced anxiety. Furthermore, using a molecular genetic approach, we demonstrated that chronic pain elevated the excitability of a specific subpopulation of BNST neurons, which express cocaine- and amphetamine-regulated transcript (CART). The elevated excitability of CART-positive neurons caused the increased inhibitory inputs to LH-projecting BNST neurons, thereby inducing anxiety-like behavior. These findings shed light on how chronic pain induces psychiatric disorders, characterized by maladaptive anxiety.

Although dysfunction of the mesolimbic dopaminergic system has been implicated in chronic pain, the underlying mechanisms remain to be elucidated. We hypothesized that increased inhibitory inputs to the neuronal pathway from the dorsolateral bed nucleus of the stria terminalis (dlBNST) to the ventral tegmental area (VTA) during chronic pain may induce tonic suppression of the mesolimbic dopaminergic system. To test this hypothesis, male Sprague Dawley rats were subjected to spinal nerve ligation to induce neuropathic pain and then spontaneous IPSCs (sIPSCs) were measured in this neuronal pathway. Whole-cell patch-clamp electrophysiology of brain slices containing the dlBNST revealed that the frequency of sIPSCs significantly increased in VTA-projecting dlBNST neurons 4 weeks after surgery. Next, the role of corticotropin-releasing factor (CRF) signaling within the dlBNST in the increased sIPSCs was examined. CRF increased the frequency of sIPSCs in VTA-projecting dlBNST neurons in sham-operated controls, but not in chronic pain rats. By contrast, NBI27914, a CRF type 1 receptor antagonist, decreased the frequency of sIPSCs in VTA-projecting dlBNST neurons in the chronic pain rats, but not in the control animals. In addition, histological analyses revealed the increased expression of CRF mRNA in the dlBNST. Finally, bilateral injections of NBI27914 into the dlBNST of chronic pain rats activated mesolimbic dopaminergic neurons and induced conditioned place preference. Together, these results suggest that the mesolimbic dopaminergic system is tonically suppressed during chronic pain by enhanced CRF signaling within the dlBNST via increased inhibitory inputs to VTA-projecting dlBNST neurons.SIGNIFICANCE STATEMENTThe comorbidity of chronic pain and depression has long been recognized. Although dysfunction of the mesolimbic dopaminergic system has been implicated in both chronic pain and depression, the underlying mechanisms remain to be elucidated. Here, we show that the inhibitory inputs to the neuronal pathway from the dorsolateral bed nucleus of the stria terminalis (dlBNST) to the ventral tegmental area increase during chronic pain. This neuroplastic change is mediated by enhanced corticotropin-releasing factor signaling within the dlBNST that leads to tonic suppression of the mesolimbic dopaminergic system, which may be involved in the depressive mood and anhedonia under the chronic pain condition.

Background A method that promotes the retrieval of lost long-term memories has not been well established. Histamine in the central nervous system is implicated in learning and memory, and treatment with antihistamines impairs learning and memory. Because histamine H3 receptor inverse agonists upregulate histamine release, the inverse agonists may enhance learning and memory. However, whether the inverse agonists promote the retrieval of forgotten long-term memory has not yet been determined. Methods Here, we employed multidisciplinary methods, including mouse behavior, calcium imaging, and chemogenetic manipulation, to examine whether and how the histamine H3 receptor inverse agonists, thioperamide and betahistine, promote the retrieval of a forgotten long-term object memory in mice. In addition, we conducted a randomized double-blind, placebo-controlled crossover trial in healthy adult participants to investigate whether betahistine treatment promotes memory retrieval in humans. Results The treatment of H3 receptor inverse agonists induced the recall of forgotten memories even 1 week and 1 month after training in mice. The memory recovery was mediated by the disinhibition of histamine release in the perirhinal cortex, which activated the histamine H2 receptor. Histamine depolarized perirhinal cortex neurons, enhanced their spontaneous activity, and facilitated the reactivation of behaviorally activated neuronal ensembles. A human clinical trial revealed that treatment of H3 receptor inverse agonists is specifically more effective for items that are more difficult to remember and subjects with poorer performance. Conclusions These results highlight a novel interaction between the central histamine signaling and memory engrams.

BACKGROUND: The authors sought to quantify the results of clinical and radiological analyses of extreme lateral interbody fusion (XLIF) plus percutaneous pedicle screw (PPS) fixation for patients with lumbar spinal stenosis (LSS) by focusing on the distinct mechanism of indirect decompression. METHODS: Data obtained from a total of 37 patients with 47 surgical sites were retrospectively analyzed. Clinical outcomes for all patients were evaluated using the Japanese Orthopaedic Association (JOA) score and the improvement rate of the JOA score. Preoperative and postoperative magnetic resonance images were used to measure the transverse areas of both the dural sac (DS area) and ligamentous flavum (LF area) in the axial sections and the length of the intervertebral disc bulge (DB length) in sagittal sections. Then, the rate of change (RC) of the DS area (RC-DS), the RC of the LF area (RC-LF), and the RC of the DB length (RC-DB) from the preoperative period to the postoperative period were calculated. Furthermore, we divided all surgical sites into the small expansion group (SE group; RC-DS

Information from direct experience and observation of others is integrated in the brain to enable appropriate responses to environmental stimuli. Fear memory can be acquired by observing a conspecific’s distress. However, it remains unclear how prior fear observation affects self-experienced fear learning. In this study, we tested whether prior observation of a conspecific receiving contextual fear conditioning affects subsequent self-experienced fear conditioning and how neuronal ensembles represent the integration of the observation and self-experience. Test mice observed demonstrator mice experiencing fear conditioning on day 1 and directly experienced fear conditioning on day 2. Contextual fear memory was tested on day 3. The prior observation of fear conditioning promoted subsequent self-experienced fear conditioning in a hippocampus-dependent manner. We visualized hippocampal neurons that were activated during the observation and self-experience of fear conditioning and found that self-experienced fear conditioning preferentially activated dorsal CA1 neurons that were activated during the observation. When mice observed and directly experienced fear conditioning in different contexts, preferential reactivation was not observed in the CA1, and fear memory was not enhanced. These findings indicate that dorsal CA1 neuronal ensembles that were activated during both the observation and self-experience of fear learning are implicated in the integration of observation and self-experience for strengthening fear memory.

Motivational states are complex and consist of cognitive, emotional, and physiological components controlled by a network across multiple brain regions. An integral component of this neural circuitry is the bed nucleus of the stria terminalis (BNST). Here, we identified a subpopulation of neurons within BNST expressing the gene prepronociceptin (PnocBNST), that can modulate the rapid changes in physiological arousal that occur upon exposure to stimuli with motivational salience. Using in vivo two-photon calcium imaging we found that excitatory responses from individual PnocBNST neurons directly corresponded with rapid increases in pupillary size and occurred upon exposure to both aversive and rewarding odors. Furthermore, optogenetic activation of these neurons increased pupillary size, but did not alter approach/avoidance or locomotor behaviors. These findings suggest that excitatory responses in PnocBNST neurons encode rapid arousal responses irrespective of tested behaviors. Further histological, electrophysiological, and single-cell RNA sequencing data revealed that PnocBNST neurons are composed of genetically and anatomically identifiable subpopulations that can be further investigated. Taken together, our findings demonstrate a key role for a PnocBNST neuronal ensemble in encoding the rapid arousal responses that are triggered by motivational stimuli.

Aims Thermoregulatory responses in homeothermic animals, including humans, are classified into involuntary autonomous and voluntary behavioral thermoregulatory responses. Although behavioral thermoregulatory responses are probably driven by positive (pleasant) and/or negative (unpleasant) emotions, the neuronal mechanisms underlying the induction of negative emotions by hot and cold environments remain poorly understood. The bed nucleus of the stria terminalis is a brain region implicated in stress responses and negative emotions, such as fear, anxiety, and aversion. Various stimuli that cause negative emotions, such as immobilization stress, fox odor, gastric distension, and inflammatory pain, increase noradrenaline release in the rat bed nucleus of the stria terminalis, especially in the ventral bed nucleus of the stria terminalis. It has been reported that the negative emotional component of pain is mediated by noradrenergic neurotransmission in the ventral bed nucleus of the stria terminalis. However, the role of intra‐ventral bed nucleus of the stria terminalis noradrenergic neurotransmission in the induction of negative emotion by exposure to hot and cold environments remains to be elucidated. For the first step to address this issue, the effects of hot and cold environments on noradrenaline release in the ventral bed nucleus of the stria terminalis were examined. Methods In vivo microdialysis analyses in unanesthetized, freely moving male Sprague‐Dawley rats were performed to examine hot and cold environments‐induced noradrenaline release in the ventral bed nucleus of the stria terminalis. Results Exposure to hot (38°C) and cold (8°C) environments significantly increased noradrenaline release in the ventral bed nucleus of the stria terminalis. Conclusions The results suggest that exposure to hot and cold environments enhances noradrenergic neurotransmission in the ventral bed nucleus of the stria terminalis, which may induce negative emotion, and thereby drive avoidance behaviors, that is, escape from hot and cold environments., Intra‐ventral bed nucleus of the stria terminalis noradrenergic transmission is reported to mediate negative emotions. In the present study using an in vivo microdialysis technique, we demonstrated that hot (38°C) and cold (8°C) environments increased noradrenaline release in the ventral bed nucleus of the stria terminalis. Noradrenergic transmission may drive escape behaviors from hot and cold environments.

In daily medical practice, scapular pain associated with cervical radiculopathy is commonly seen prior to the onset of pain of the upper extremity; however, the cause of the scapular pain is uncertain. We report a case of herpes zoster with simultaneous skin rash in both the upper extremity and interscapular region, which corresponds to the painful scapular region in case of C8 cervical radiculopathy. A 71-year-old healthy woman complained of shoulder and scapular pain followed by a blistering skin rash on both the ulnar side of her upper extremity and intrascapular region on the right side. She was diagnosed with herpes zoster and was prescribed amenamevir as oral treatment with vidarabine ointment. After 1 year, she still had mild causalgia on her III–V fingers and needed oral treatment with pregabalin. To our knowledge, this is the first case report of a herpes zoster rash in the upper extremity and intrascapular region simultaneously. We speculate that the rash in the two regions is caused by the varicella zoster virus (VZV) traveling from the same ganglion, probably the C8 ganglion, considering the dermatome of the rash area in the upper extremity and the intrascapular region correspond to the innervation zone of the medial branches of the dorsal ramus of the cervical nerve root, which resembles the scapular region in case of cervical radiculopathy. This phenomenon implies the mechanism of scapular pain is related to cervical radiculopathy. Further case reports are needed to confirm this.

Lumbar spinal stenosis (LSS) is a pathological condition in which degenerative changes in the lumbar spine lead to spinal canal narrowing and compression of the dural tube and spinal nerve roots. Most patients with LSS have accompanying lumbar spondylosis with or without spondylolisthesis based on the aging process, and some experience regional low back pain due to spinal instability. Surgical treatment is indicated for patients with LSS who do not respond to conservative therapy. Conventional treatments are decompression alone and decompression plus spinal fusion using a pedicle screw and rod system with bone graft materials or a combination of anterior and posterior fusion. In terms of decompressive procedure, insufficient decompression can cause recurrence of stenosis after surgery; however, too much removal of the posterior elements and excessive facetectomy can cause postoperative spinal instability of the operated level. Iatrogenic fracture of the pars interarticularis due to massive laminectomy during surgery may result in isthmic spondylolisthesis. To avoid postoperative instability or to reduce regional low back pain due to degenerative spinal instability, additional spinal fusion is a promising strategy. Between 2002 and 2007 in the United States, 4,699 (79.4%) of 5,915 patients with stenosis plus spondylolisthesis were treated with decompression plus spinal fusion, whereas 19,699 (78.6%) of 25,060 patients with stenosis alone were treated with decompression alone (1). Whether additional spinal fusion is essential for LSS with low-grade spondylolisthesis remains controversial, but there is no doubt that spinal fusion is the gold standard operation for LSS.

Sacral fatigue fractures in the young population usually occur due to repetitious physical strain from vigorous athletic activity; they are extremely rare in those younger than 13 years without intense physical activity. We report a case of sacral fatigue fracture in a healthy elementary school girl without any history of trauma or excessive athletic activity. A healthy 11-year-old girl experienced sudden low back pain upon standing after a short break in a normal running exercise for physical education at school. At her first orthopaedic visit, she was unable to walk independently and was limping on her left leg. Neurological examination revealed that the left straight leg raising test was positive at 80 degrees with associated left buttock pain but without motor and sensory deterioration. Radiological examination showed no obvious fractures in the lumbar vertebrae or pelvis. Magnetic resonance imaging demonstrated high intensity signal changes on the short tau inversion recovery image of the left ala, and sacral fatigue fracture in the left ala was diagnosed. She was instructed to rest at home and allowed minimal walking with a crutch under endurable pain for 4 weeks. Within 3 weeks, her low back pain gradually reduced, and after 4 weeks, she could walk independently without gait pain. Sacral fatigue fractures should be considered in the diagnosis of young patients who present with unexplained low back pain.

Center for Information and Neural Networks, Suita City,Osaka 565-0871, JapanSummaryThe frontal association cortex (FrA) is implicated in higherbrain function [1]. Aberrant FrA activity is likely to beinvolved in dementia pathology [2–4]. However, the func-tional circuits both within the FrA and with other regionsare unclear. A recent study showed that inactivation of theFrAimpairsmemoryconsolidationofanauditoryfearcondi-tioninginyoungmice[5].Inaddition,dendriticspineremod-eling of FrA neurons is sensitive to paired sensory stimulithat produce associative memory [5]. These findings sug-gest that the FrA is engaged in neural processes criticalto associative learning. Here we characterize stimulus inte-gration in the mouse FrA during associative learning. Weexperimentally separated contextual fear conditioning intocontext exposure and shock, and found that memory forma-tion requiresprotein synthesisassociatedwith bothcontextexposure and shock in the FrA. Both context exposure andshock trigger Arc, an activity-dependent immediate-earlygene, expression in the FrA, and a subset of FrA neuronswas dually activated by both stimuli. In addition, we foundthat the FrA receives projections from the perirhinal (PRh)and insular (IC) cortices and basolateral amygdala (BLA),which are implicated in context and shock encoding [6–8].PRh and IC neurons projecting to the FrA were activatedby context exposure and shock, respectively. Arc expres-sion in the FrA associated with context exposure and shockdepended on PRh activity and both IC and BLA activities,respectively.ThesefindingsindicatethattheFrAisengagedinstimulusintegrationandcontributestomemoryformationin associative learning.ResultsThe Frontal Association Cortex Is Required for MemoryFormation in Contextual Fear ConditioningAsamodelforassociativelearning,weusedacontextualfear-conditioning task, which establishes an association betweencontext and shock. To test whether the frontal associationcortex (FrA) is involved in memory formation in contextualfear conditioning, we infused (2R)-amino-5-phosphonovalericacid(APV),anN-methyl-D-aspartate(NMDA)receptorantago-nist, anisomycin, a protein synthesis inhibitor, or vehicle intothe FrA. APV and anisomycin were infused 30 min prior to, orimmediately after, contextual fear conditioning, respectively(experiment 1; Figures 1A and 1B). Contextual fear memorywas assessed by measuring the percentage of freezing timein the conditioning context 1 day after conditioning. BothAPV and anisomycin infusions disrupted freezing behavior.When anisomycin infusions were administered into the dorso-medial prefrontal cortex, which is close to the FrA (experiment2; Figure S1A available online), freezing behavior was compa-rable to that of mice administered vehicle infusions (Fig-ureS1B).TheseresultsindicatethatNMDAreceptoractivationandproteinsynthesisintheFrAarerequiredforcontextualfearconditioning.ProteinSynthesisintheFrAIsRequiredforEncodingBothContext and ShockWe aimed to determine whether the FrA encodes context,shock, or both. To this end, we separated 10 min of contextexposureandimmediateshockbya1-dayintervalandinfusedanisomycinintotheFrAaftereithercontextexposureorshock(experiment 3; Figures 1C and 1D). Anisomycin infusions intothe FrA immediately after both context exposure and immedi-ate shock disrupted freezing behavior during the test. How-ever, when anisomycin infusions were administered into theFrA 6 hr after context exposure (Figure 1E), freezing behaviorwas comparable to that of mice administered vehicle infu-sions. These results indicate that protein synthesis in the FrAis required for encoding both context and shock.FrA Neurons Receive Convergent Information RegardingContext and Shock during Fear ConditioningBecause protein synthesis in the FrA is required for encodingboth context and shock, we hypothesized that paired stimuliconverge in a subset of FrA neurons to potentially contributeto the memory trace. To visualize stimulus convergence, weanalyzed the temporal dynamics of nuclear versus cyto-plasmic Arc localization by fluorescent in situ hybridization[9]. Arc is an activity-dependent immediate-early gene thatis essential for synaptic plasticity and long-term memory[10–13]. Transcribed Arc mRNA first appears in neuronalnuclei, and processed Arc mRNA then accumulates in thecytoplasm. Thus, an analysis of the subcellular localization ofArc enabled us to identify active neuronal ensembles duringtwo behavioral tasks [9, 14, 15]. We first examined the timecourse of the nuclear and cytoplasmic Arc signal after neuralactivity in the FrA. Mice were exposed to a context for 5 minand sacrificed either immediately or 30 min later (experiment4; Figures S2A and S2B). We observed more nuclear Arc

Objectives Agricultural use of neonicotinoid (NEO) insecticides has been increasing in recent years, but their biological monitoring methods have been scarcely reported. In this study, we developed and validated a rapid and sensitive method for quantifying urinary NEO concentrations using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods After phosphate-induced acidification of a urine sample, urinary NEOs were trapped by a solid-phase extraction column and eluted with methanol for acetamiprid, imidacloprid, thiacloprid, thiamethoxam, clothianidin and dinotefuran and with an acetonitrile and methanol solution (1:1, v/v) containing 5% NH3 for nitenpyram. A separation analysis was performed by LC-MS/MS within 10 minutes for the sample. This method was applied to first morning urine obtained from 52 Japanese (40.9 ± 10.5 years old, mean ± standard deviation) without occupational NEO exposure. Results The linear dynamic ranges and their limit of quantification (LOQ, signal to noise ratio=10) levels were 0.3-20 or 50 µg/l (r=0.998-0.999) and 0.05-0.36 µg/l, respectively. The absolute recovery was 64-95%, and the intra- and inter-day precisions were less than 16.4% (relative standard deviation, %RSD). This method was successfully applied for analysis of NEOs in human urine samples obtained from 52 adults. The frequencies of individuals who showed more than LOD levels was above 90% for imidacloprid, thiamethoxam, clothianidin and dinotefuran, more than 50% for acetamiprid and thiacloprid and 29% for nitenpyram. Conclusions These results indicated that our new method could be applied to biological monitoring of NEO exposure even at environmental exposure levels in Japanese adults without occupational spraying histories.

In 2012, we established the Center for Regenerative Medicine in Dokkyo Medical University Hospital, and are now preparing the necessary equipment and preclinical evidence for cell therapy. Liposuction is a commonly used procedure in plastic surgery and the lipoaspirate is discarded as a medical waste. However, the lipoaspirate is known to contain abundant mesenchymal stem cells, and thus, it is currently one of the most feasible options of regenerative medicine. Several ongoing clinical trials of cell therapy (in Japan and overseas as well) are based on the Celution^ system, an automated cell-processing machine utilizing lipoaspirate. The merit of using such a machine is that collected cells are so abundant that it is not necessary to expand the cell number in a Cell Processing Center. Moreover, there are fewer risks of bacterial infection because the system is isolated from the ambient dust and operated automatically. Our group is now planning respective cell therapies for breast reconstruction after mastectomy, urinary incontinence and ischemic cardiovascular diseases, and all these protocols will be based on the Celution^ system. Here, we report the initial test run of Celution^ to confirm its safety and reproducibility.

Historically, Hansen’s disease patients suffered from discrimination because their physical features changed due to the bacterium Mycobacterium leprae (M. leprae) and made them “ugly” in the eyes of society. Former Japanese governments saw them as a national disgrace and forced them to reside in leprosaria. Since the law requiring isolation continued after the silver bullet was developed, survivors could not leave the leprosaria and return to society. Currently, survivors’ average age is 82 and they live in 13 national sanatoriums. When they pass away, the history of Hansen’s disease in Japan will end, so we must record their experiences. We conducted qualitative and inductive studies with survivors. In this chapter, we reconstruct them from the perspective of bioethics and propose several theories surrounding them: (1) How former leprosaria and medical administrations in Japan threatened bioethical principles; (2) the wisdom of aging survivors, who lived through extreme situations, and what real restoration of their rights might look like; and (3) the ethical dilemmas of how we will care for the survivors—who have multiple severe sequelae—until they all pass away. Finally, we will intro

The present study reports the beneficial effects of an anti-mouse interleukin-6 (IL-6) receptor antibody (MR16-1) on neuropathic pain in mice with spinal cord injury (SCI). Following laminectomy, contusion SCI models were produced using an Infinite Horizon (IH)-impactor. MR16-1 was continuously injected for 14 days using Alzet osmotic pumps. A mouse IL-6 ELISA kit was then used to analyze IL-6 levels in the spinal cord tissue between 12 and 72 h after injury. Motor and sensory functions were evaluated each week using the Basso Mouse Scale (BMS), plantar von Frey and thermal threshold tests. Histological examinations were performed 42 days after SCI. Between 24 and 72 h after SCI, the expression levels of IL-6 were significantly decreased in the MR16-1 treated group. Six weeks after surgery, the BMS score of the MR16-1-treated group indicated significant recovery of neurological functions. MR16-1-treated mice in the SCI group exhibited lower paw withdrawal thresholds in the plantar von Frey and thermal tests, which were used to evaluate allodynia. MR16-1 treatment significantly increased the area of Luxol fast blue-stained tissue, representing spared myelin sheaths. These results indicate that the continuous inhibition of IL-6 signaling by MR16-1 between the early and sub-acute phases following SCI leads to neurological recovery and the suppression of hyperalgesia and allodynia. Overall, our data suggest that the inhibition of severe inflammation may be a promising neuroprotective approach to limit secondary injury following SCI and that an anti-IL-6 receptor antibody may have clinical potential for the treatment of SCI.

The aim of this study was to develop a method for quantitative measurement of urinary metabolites of pyrethroid (PYR) insecticides, trans-chrysanthemumdicarboxylic acid (CDCA) and 3-phenoxybenzoic acid (3-PBA), extracted from disposable diapers. This study was approved by the university ethics committees, and informed consent was obtained from all the parents for their children and from adult volunteers. After extraction of PYR metabolites in the absorber of diapers with 5 ml acetone, the metabolites in the eluents were extracted with tert-butyl methyl ether, derivatized with 1,1,1,3,3,3-hexafluoroisopropanol and analyzed by gas chromatography-mass spectrometry. The limits of quantitation (LOQs) were 0.55 μg/l for CDCA and 0.09 μg/l for 3-PBA in 2 ml urine extracted from diapers. Within-series and between-day precisions were