Melting of a single ice microparticle on exposure to focused near-IR laser beam to yield a supercooled water droplet.

We observed for the first time that a single ice microparticle supported on a substrate melted photothermally to form a supercooled water droplet on exposure to tightly focused illumination with a 1064-nm laser beam that generated a point heat source. In situ Raman micro-spectroscopy clearly showed the formation of liquid water at the expense of ice. The observation of this melting is only possible when the experiment is performed with micrometer-sized ice particles. A previous attempt to melt millimeter-sized ice through photothermal heating of gold nanoaggregates fell short of expectations because only vapor formation, rather than liquid water formation, has been postulated. Our observation is significant because thermal confinement in a microscale compartment using a water-air interface as a heat-insulated wall can achieve particle temperatures above the melting point of water, whereas, in an unlimited space of ice, heat transfer from the heating center to the surroundings causes steep temperature decays, resulting in limited temperature increase.

The 10 Hz dynamic response of a fluid-filled pressure monitoring system is a novel alternative to the fast flush test and indicative of unacceptable systolic pressure overshoot.

The standard method for qualitatively evaluating the dynamic response is to see if the gain of the amplitude spectrum curve approaches 1 (input signal = output signal) over the frequency band of the blood pressure waveform. In a previous report, Watanabe reported that Gardner's natural frequency and damping coefficient, which are widely used as evaluation methods, do not reflect the dynamic response of the circuit. Therefore, new parameters for evaluating the dynamic response of pressure monitoring circuits were desired. In this study, arterial pressure catheters with length of 30, 60, 150, and 210 cm were prepared, and a blood pressure wave calibrator, two pressure monitors with analog output and a personal computer were used to analyze blood pressure monitoring circuits. All data collection and analytical processes were performed using step response analysis program. The gain at 10 Hz was close to 1 and the systolic blood pressure difference was small in the short circuits (30 cm, 60 cm), and the gain at 10 Hz was 1.3-1.5 in the 150 cm circuit and over 1.7 in the 210 cm circuit. The difference in systolic blood pressure increased in proportion to the length of the circuit. It could also be inferred that the gain at 10 Hz should be less than 1.2 to meet a clinically acceptable blood pressure difference. In conclusion, the gain at 10 Hz is sufficiently useful as an indicator to determine the correct systolic blood pressure.

IL-6 Plays a Critical Role in Stromal Fibroblast RANKL Induction and Consequent Osteoclastogenesis in Ameloblastoma Progression.

Ameloblastoma (AB) is the most common benign, epithelial odontogenic tumor that occurs in the jawbone. AB is a slow-growing, benign epithelial tumor but shows locally invasive growth, with bone resorption or recurrence if not adequately resected. From these points of view, understanding the mechanism of AB-induced bone resorption is necessary for better clinical therapy and improving patients' quality of life. In bone resorption, osteoclasts play critical roles, and RANKL is a pivotal regulator of osteoclastogenesis. However, the source of RANKL-expressing cells in the AB tumor microenvironment is controversial, and the mechanism of osteoclastogenesis in AB progression is not fully understood. In this study, we investigated the distribution of the RNA expression of RANKL in AB specimens. We found that PDGFRα- and S100A4-positive stromal fibroblasts expressed RANKL in the AB tumor microenvironment. Moreover, we analyzed the mechanisms of osteoclastogenesis in the AB tumor microenvironment using the human AB cell line AM-1 and a human primary periodontal ligament fibroblast cells. The results of histopathologic and in vitro studies clarified that the interaction between AB cells and stromal fibroblasts upregulated IL-6 expression and that AB cells induced RANKL expression in stromal fibroblasts and consequent osteoclastogenesis in AB progression.

Inhibition of NPR1 Leads to Shoot Growth Improvement under Low-Calcium Conditions in Arabidopsis.

Under low-Ca conditions, plants accumulate salicylic acid (SA) and induce SA-responsive genes. However, the relationship between SA and low-Ca tolerance remains unclear. Here, we demonstrated that the inhibition or suppression of nonexpressor of pathogenesis-related 1 (NPR1) activity, a major regulator of the SA signaling pathway in the defense response, improves shoot growth under low-Ca conditions. Furthermore, mutations in phytoalexin-deficient 4 (PAD4) or enhanced disease susceptibility 1 (EDS1), which are upstream regulators of NPR1, improved shoot growth under low-Ca conditions, suggesting that NPR1 suppressed growth under low-Ca conditions. In contrast, growth of SA induction-deficient 2-2 (sid2-2), which is an SA-deficient mutant, was sensitive to low Ca levels, suggesting that SA accumulation by SID2 was not related to growth inhibition under low-Ca conditions. Additionally, npr1-1 showed low-Ca tolerance, and the application of tenoxicam-an inhibitor of the NPR1-mediated activation of gene expression-also improved shoot growth under low Ca conditions. The low-Ca tolerance of double mutants pad4-1, npr1-1 and eds1-22 npr1-1 was similar to that of the single mutants, suggesting that PAD4 and EDS1 are involved in the same genetic pathway in suppressing growth under low-Ca conditions as NPR1. Cell death and low-Ca tolerance did not correlate among the mutants, suggesting that growth improvement in the mutants was not due to cell death inhibition. In conclusion, we revealed that NPR1 suppresses plant growth under low-Ca conditions and that the other SA-related genes influence plant growth and cell death.

Utility of a haemoglobin test of gingival crevicular fluid: A multicentre, observational study.

OBJECTIVE: The purpose of this study was to verify the accuracy and utility of clinical parameters (plaque index, gingival crevicular fluid volume, probing depth, clinical attachment level, bleeding on probing and gingival index) and biochemical parameters (aspartate aminotransferase, protein and haemoglobin) in a longitudinal analysis during the supportive periodontal therapy period. SUBJECTS AND METHODS: A total of 279 test sites of 128 patients were investigated clinically and biochemically. After the first examination of clinical and biochemical parameters, periodontal support treatments were administered immediately and performed once every three months up to the second examination. RESULTS: All of the clinical and biochemical parameters were significantly lower at the second examination than at the first, except for the plaque index and bleeding on probing. Of these parameters, in particular, aspartate aminotransferase and haemoglobin in the gingival crevicular fluid were significantly reduced compared to those of the first examination in both the ≤4 and ≥5 mm probing depth groups, and they clearly suggested that periodontitis tended to recover. CONCLUSION: Adding the haemoglobin test to the bleeding on probing test strongly improves the accuracy of measurement of clinical parameters after periodontal treatment.

Effect of remimazolam on intraoperative neurophysiology monitoring of visual-evoked potential: a case series.

There are very few reports on the effects of benzodiazepines such as midazolam and diazepam on intraoperative visual-evoked potential (VEP), and there is no report on the effect of remimazolam at all. Five patients underwent neurosurgery using VEP monitoring for avoiding surgical injury to the optic nerve. In all cases, drug administration was based on actual body weight. General anesthesia was induced with propofol and remifentanil, and then maintained with propofol at target concentrations of 2.7-3.5 µg/ml for maintaining bispectral index (BIS) between 40 and 60. After resection of the tumor under stable VEP, we discontinued propofol immediately followed by infusion of remimazolam at 12 mg/kg/h for a few seconds, then reduced to 1 mg/kg/h. After a time, when blood levels of remimazolam appeared to be stable, VEP was monitored again and compared to controls. In all cases, we were able to confirm that there was reproducibility. Remimazolam may provide a comparable quality of anesthesia to that of existing drugs for VEP in neurosurgery.

The intraoperative motor-evoked potential when propofol was changed to remimazolam during general anesthesia: a case series.

Remimazolam is a short-acting benzodiazepine that was approved for clinical use in 2020. We report three patients who underwent surgery for cerebral and spinal cord tumors, in whom transcranial electrical stimulation-motor-evoked potential (TES-MEP) was successfully monitored under general anesthesia with remimazolam. During total intravenous anesthesia with propofol at a target concentration of 2.7 - 3.5 µg/mL and 0.1 - 0.35 µg/kg/min of remifentanil, delayed awakening, bradycardia, and hypotension during propofol anesthesia were expected in all three cases. With patient safety as the top priority, we considered changing the anesthetic agent. Propofol was replaced with remimazolam at a loading dose of 12 mg/kg/h for a few seconds (case 3), followed by 1 mg/kg/h for maintenance (cases 1-3). TES-MEP was recorded during propofol and remimazolam administration in all three patients. Amplitudes of TES-MEP during anesthesia with propofol and remimazolam were 461.5 ± 150 µV and 590.5 ± 100.9 µV, 1542 ± 127 µV and 1698 ± 211 µV, and 581.5 ± 91.3 µV and 634 ± 82.7 µV sequentially from Case 1. Our findings suggest that intraoperative TES-MEP could be measured when anesthesia was managed with remimazolam at 1 mg/kg/h.

αTAT1-induced tubulin acetylation promotes ameloblastoma migration and invasion.

Ameloblastoma (AB) is the most common benign epithelial odontogenic tumor occurring in the jawbone. AB is a slowly growing tumor but sometimes shows a locally invasive and an aggressive growth pattern with a marked bone resorption. In addition, the local recurrence and distant metastasis of AB also sometimes occurs, which resembles one of the typical malignant potentials. From these points of view, to understand better the mechanisms of AB cell migration or invasion is necessary for the better clinical therapy and improvements of the patients' quality of life. Microtubules in eukaryotic cells reveal the shape of hollow cylinders made up of polymerized alpha (α)- and beta (ß)-tubulin dimers and form the cytoskeleton together with microfilaments and intermediate filaments. Microtubules play important roles in cell migration by undergoing assembly and disassembly with post-translational modifications. Stability of microtubules caused by their acetylation is involved in cell migration. In this study, we investigated the expression and distribution of acetylated α-tubulin and alpha-tubulin N-acetyltransferase 1 (αTAT1), an enzyme which acetylates Lys-40 in α-tubulin, in AB specimens, and analyzed how tubulin was acetylated by αTAT1 activation in a human AB cell line, AM-1. Finally, we clarified that TGF-ß-activated kinase1 (TAK1) was phosphorylated by TGF-ß stimulation, then, induced tubulin acetylation via αTAT1 activation, which subsequently activated the migration and invasion of AB cells.

Novel Bi-Specific Immuno-Modulatory Tribodies Potentiate T Cell Activation and Increase Anti-Tumor Efficacy.

Cancer immunotherapy has already shown significant improvements by combining different antibodies specific for distinct immune checkpoints, such as Ipilimumab and Nivolumab. Here, we tested combinatorial treatments of immunomodulatory antibodies, previously generated in our laboratory, for their effects on hPBMC activation, either upon stimulation with SEB or in co-cultures with tumor cells by cytokine secretion assays. We found that some of them showed additive or synergistic effects, and on the basis of these observations, we constructed, for the first time, four novel bispecific tribodies (TR), made up of a Fab derived from one anti-IC mAb and two scFvs derived from another mAb targeting a different IC. All four TRs cotargeting either programmed cell death protein 1 (PD-1) and Lymphocyte Activating 3 (LAG-3) or programmed death-ligand 1 (PD-L1) and LAG-3 retained binding affinity for their targets and the antagonistic effects of their parental mAbs, but some of them also showed an increased ability to induce lymphocyte activation and increased in vitro cytotoxicity against tumor cells compared to parental antibodies used either alone or in combinatorial treatments. Furthermore, none of the tribodies showed significant increased cytotoxicity on human cardiomyocytes. Considering that the tribody format reduces production costs (as only one construct provides the inhibitory effects of two antibodies), has an intermediate molecular size (100 kDa) which is well suited for both tumor penetration and an acceptable half-life, we think that these novel immunomodulatory TRBs have the potential to become precious tools for therapeutic applications, particularly in monotherapy-resistant cancer patients.

NFAT5 promotes oral squamous cell carcinoma progression in a hyperosmotic environment.

Epidermal growth factor receptor (EGFR) is highly expressed in several types of cancer cells including oral squamous cell carcinoma (OSCC). EGF/EGFR signaling is recognized as an important molecular target in cancer therapy. However, cancer cells often become tolerant to EGF/EGFR signaling-targeted therapies. In the tumor microenvironment, the tumor incites inflammation and the inflammation-derived cytokines make a considerable impact on cancer development. In addition, hyperosmolarity is also induced, but the role of osmotic stress in cancer development has not been fully understood. This study demonstrates molecular insights into hyperosmolarity effect on OSCC development and shows that NFAT5 transcription factor plays an important functional role in enhancing the oral cancer cell proliferation by inducing the EGFR translocation from the endoplasmic reticulum to the plasma membrane through increase the expression of DPAGT1, an essential enzyme for catalyzing the first committed step of N-linked protein glycosylation. These results suggest that hyperosmolarity-induced intra-nuclear translocation of NFAT5 essential for DPAGT1 activation and EGFR subcellular translocation responsible for OSCC tumor progression.

Periodontal inflammation triggers a site-specific and wide radius of calcium metabolic effects on alveolar bone.

BACKGROUND AND OBJECTIVE: There is a close relationship between inflammation and bone remodeling in the periodontium. However, previous studies have not delineated the alterations in calcium (Ca) metabolism during periodontitis progression. The aim of this current investigation was to examine Ca dynamics in alveolar bone of rats during progression of ligature-induced periodontal inflammation by using 45 Ca, which is an index of hard tissue neogenesis. MATERIAL AND METHODS: To induce periodontitis, the maxillary right first molar (M1) of 8-week-old male rats was ligated with a silk suture for 1, 3, 7, and 28 days. The left M1 was not ligated as a control. To evaluate resultant changes in bone neogenesis, 45 CaCl2 was injected intraperitoneally 24 hours before euthanasia. The left-and-right palatal mucosa, molar teeth (M1 and M2), and alveolar bone were harvested for evaluation of 45 Ca radioactivity using a liquid scintillation counter. The distribution of 45 Ca in maxillary tissues was evaluated using autoradiography (ARG). In addition, we analyzed the bone volume fraction (BV/TV) and bone mineral density (BMD) of the alveolar bone by micro-computed tomography. To investigate the number of osteoclasts and osteoblasts, tartrate-resistant acid phosphatase (TRAP) and bone-specific alkaline phosphatase (BAP) were measured by an enzymatic assay and immunohistochemistry, respectively. RESULTS: 45 Ca radioactivity in the alveolar bone of the ligature side decreased by 8% compared to the unligated control-side on day 1, whereas on day 7, it markedly increased by 33%. The 45 Ca levels in the gingival tissue and molar teeth were slightly but significantly lower than the control-side on day 1 and higher from day 3 to 28. The variation in 45 Ca levels for the alveolar bone was greater and specific compared with other tissues. Furthermore, on day 7, ARG data revealed that 45 Ca on the control side was primarily localized to the periodontal ligament (PDL) space and alveolar bone crest and barely detected in the gingival tissues and deeper parts of the alveolar bone. On the ligature side, 45 Ca disappeared from the PDL and alveolar crest, but instead was broadly and significantly increased within the deeper zones of the alveolar bone and furcation areas and distant from the site of ligature placement and periodontal inflammation. In the shallow zone of the alveolar bone, these changes in 45 Ca levels on day 7 were consistent with decreases in the bone structural parameters (BV/TV and BMD), enhanced osteoclast presence, and suppressed levels of BAP expression in osteoblasts. In contrast, the deep zone and furcation area showed that TRAP-positive cells increased, but BAP expression was maintained in the resorption lacunae of the alveolar bone. CONCLUSION: During periodontitis progression in rats, 45 Ca levels in the alveolar bone exhibited biphasic alterations, namely decreases and increases. These data indicate that periodontitis induces a wide range of site-specific Ca metabolism alterations within the alveolar bone.

Usefulness of hemoglobin examination in gingival crevicular fluid during supportive periodontal therapy to diagnose the pre-symptomatic state in periodontal disease.

OBJECTIVES: The absence of bleeding on probing (BOP) is a good predictor of disease stability. This study investigated whether detection of hemoglobin (Hb) in gingival crevicular fluid (GCF) indicates minute signs of periodontal disease, even in BOP (-) cases. MATERIALS AND METHODS: GCF was collected from gingival sulci of 152 sound maxillary and mandibular teeth from 76 patients who had entered supportive periodontal therapy (SPT) using the split-mouth design. As clinical parameters, plaque index, GCF amount, gingival index, probing depth (PD), clinical attachment level, BOP, and alveolar bone resorption ratio were then recorded. As biochemical parameters, Hb amount, alkaline phosphatase (ALP) activity, and protein amount in GCF were measured. Periodontal conditions of diseased sites (PD ≥ 4 mm, BOP (+)) and healthy sites (PD ≤ 4 mm, BOP (-)) were further classified into two groups using the Hb cutoff value determined by PD and BOP and analyzed. RESULTS: Despite being healthy, ALP activity and protein amount in sulci of the group with Hb level greater than the cutoff value were significantly higher than those in the group with Hb level less than the cutoff value (P < 0.01). CONCLUSIONS: This study indicates that Hb examination is a promising candidate marker of pre-symptomatic periodontal disease because Hb presence in GCF suggests slight tissue damage, even in healthy sites defined as BOP (-). CLINICAL RELEVANCE: Hb examination of GCF is a powerful diagnostic tool for pre-symptomatic diagnosis of periodontal disease.

Temporal and dynamic changes in gingival blood flow during progression of ligature-induced periodontitis.

OBJECTIVES: To evaluate temporal changes in gingival blood flow (GBF) during progression of periodontitis in rats using a laser Doppler flowmeter (LDF) approach and to characterize morphological and biochemical features in the periodontium associated with GBF. MATERIALS AND METHODS: Forty-two Wistar rats were divided into a ligature-induced periodontitis group and a control group. To induce periodontitis, ligatures were tied around maxillary first molars bilaterally. GBF was measured in palatal gingiva at pretreatment and following ligature placement after 30 min, 1, 3, 7, 14, 21, and 28 days using LDF with a non-contact probe. Bone loss and gene expression in gingival tissues were assessed using micro-computed tomography (µCT) and quantitative polymerase chain reaction (PCR), respectively. Immunostaining for vascular endothelial growth factor (VEGF) in the maxilla was also histologically evaluated. RESULTS: GBF in the ligature group increased significantly compared with the control group 30 min after ligation. However, on days 3 and 7, GBF decreased in the ligature group. Also, after day 10, there was no difference in GBF between groups. The levels of alveolar bone loss, gene expression (interleukin-6, cluster of differentiation-31, VEGF-A, and lymphatic vessel endothelial hyaluronan receptor-1), and immunostained VEGF-positive vessels correlated well with changes in GBF. CONCLUSION PROGRESSION OF PERIODONTITIS: In rats was associated with a triphasic pattern of GBF, consisting of a short initial increase, followed by a rapid decrease, and then a gradual plateau phase.

Effect of adding vasopressin on the distribution of lidocaine in tissues, anesthetic action, and circulatory dynamics.

We examined whether vasopressin affects the distribution, anesthesia duration, and circulatory dynamics of lidocaine. Blood flow was measured after injecting 0.003, 0.03, or 0.3 U/mL vasopressin and 2% lidocaine (L) to the upper lip of rats. Radioactivity and distribution of 14C-labeled L (CL) in the palate, palatal mucosa, maxilla bone, and blood was measured by autoradiography after injecting CL and CL + 0.03 U/mL vasopressin. To evaluate anesthesia duration, somatosensory-evoked potentials, blood pressure, and pulse rate were measured after L, 0.03 U/mL vasopressin, and L + 0.03 U/mL vasopressin injection to the palatal mucosa. Blood flow from 10 to 60 min was significantly lower with 0.03 U/mL vasopressin and L + 0.03 U/mL vasopressin than with L. Radioactivity in the palatal mucosa and maxilla bone was significantly higher at 5-60 min and 2-60 min with CL + 0.03 U/mL vasopressin than with CL. Blood radioactivity reached the maximum at 0.5 and 50 min with CL and CL + 0.03 U/mL vasopressin, respectively. Autoradiogram showed higher distribution with CL + 0.03 U/mL vasopressin than CL. Peak-to-peak amplitude 30-60 min was significantly lower with L + 0.03 U/mL vasopressin than with L. Lidocaine did not affect blood pressure and pulse rate with 0.03 U/mL vasopressin-only or combined with 2%-lidocaine. Topical 0.03 U/mL vasopressin injection reduced the tissue blood flow, promoted the localization and retention, and extended the anesthesia duration of lidocaine, leaving circulatory dynamics unaffected.

Effect of vasopressin as a local anesthetic in mice.

We sought to elucidate how the local administration of mepivacaine hydrochloride and vasopressin via the tail affects the peripheral blood flow volume, tissue dynamics, and mepivacaine's anesthetic effect in mice. Two-hundred and twenty-six male ICR mice were used in this study. Blood flow was measured after administering mepivacaine alone or mepivacaine with either 0.03, 0.3, or 3.0 U/mL vasopressin or 10 µg/mL epinephrine via the tail tissue. The tail tissue and blood dynamics were measured using 3H-labeled mepivacaine hydrochloride with vasopressin or epinephrine. The compound nerve action potential (CNAP) was measured to clarify the anesthetic effect after administering mepivacaine with 0.3 U/mL vasopressin. The statistical methods employed were Steel-Dwass test, Mann-Whitney U test, Dunnett's test, and Tukey test. P < 0.05 indicated statistical significance. The results revealed that the local administration of ≥ 0.03 U/mL vasopressin reduced local blood flow and prolonged 3H-M localization in the tail tissue in a concentration-dependent manner. Addition of 0.3 U/mL vasopressin enhanced and prolonged the anesthetic effect of mepivacaine. The findings suggest that adding vasopressin to a local anesthetic regimen may be effective, and thus it could be applied as a vasoconstrictor.

Impact of dexmedetomidine on the tissue distribution, anesthetic action, and hemodynamic effects of mepivacaine.

The present study investigated the regional blood flow, tissue distribution, local anesthetic action, and hemodynamic effects of mepivacaine containing dexmedetomidine hydrochloride (DEX) in rats. Blood flow was measured after injection of 0.5% mepivacaine (M group), 12.5 µg/ml DEX (D group), or 0.5% mepivacaine containing 12.5 µg/ml DEX (DM group) into the upper lip. Mepivacaine distribution was autoradiographically observed in maxillary bone resected after injection of 0.5% 3H-mepivacaine (HM group) or 0.5% 3H-mepivacaine containing 12.5 µg/ml DEX (DHM group) into the palatal mucosa adjacent to the right maxillary first molar. Radioactivity was also measured using a liquid scintillation counter. SEP were measured to analyze anesthetic action. Blood pressure and heart rate were measured to compare hemodynamic effect. The addition of DEX significantly decreased blood flow compared to M group from 10 to 60 min after injection. The addition of DEX significantly increased the amount of radioactivity compared to HM group in the palatal mucosa from 5 to 60 min after injection and in the body of the maxilla from 2 to 60 min after injection. Maximum blood radioactivity was measured at 5 min after injection in HM group and 50 min after injection in DHM group. The addition of DEX significantly decreased peak-to-peak amplitudes compared to M group until 60 min after injection. No significant hemodynamic differences were observed. DEX enhances the action of mepivacaine in reducing regional blood flow prolongs its tissue retention, and increases the local anesthetic action without affecting hemodynamics on local administration.

Effects of experimental periodontitis on the metabolic system in rats with diet-induced obesity (DIO): an analysis of serum biochemical parameters.

Recent studies have shown that periodontitis accelerates the progression of obesity-associated metabolic diseases. Thus, we examined the influence of periodontitis on serum biochemical parameters of metabolic disease in a diet-induced obesity (DIO) rat. First, we established the DIO model using ten male rats fed with either basal diet (lean group) or high-fat diet (DIO group) for 12 weeks. Second, to examine the interaction between periodontitis and obesity, we divided 24 DIO rats into the following four groups. (1) Porphyromonas gingivalis (Pg) group was applied with Pg around the maxillary first molar (M1). (2) Ligature group was applied with ligature placement around M1. (3) Ligature/Pg group was treated with both ligature placement and Pg. (4) Control was non-treatment group. Serum biochemical parameters and maxillary histopathology were evaluated at 12 weeks. The DIO model demonstrated significant increases in body weight, serum insulin, alanine aminotransaminase (ALT) levels, and insulin resistance (HOMA-IR) compared to the lean group. In the DIO ligature and ligature/Pg groups, alveolar bone resorption and inflammatory cell infiltration were significantly increased compared to the control. Serum levels of fasting glucose, lactate dehydrogenase, and uric acid were also significantly higher, while the liver damage markers ALT and aspartate aminotransferase were only higher in ligature/Pg group. However, we observed no significant differences between the Pg group and Control. The present study suggested a possibility that periodontitis induced by ligature placement changed serum metabolic parameter regarding organs such as the liver in DIO rat.

Efficacy of intraoperative direct electrical stimulation of the spinal root and measurement of distal motor latency in lumbar spinal stenosis.

PURPOSE: The measurement of distal motor latency (DML) is an established method for diagnosing entrapment peripheral neuropathy. DML can also serve as an index for disease severity and prognosis. We considered that measuring DML could be useful in estimating the severity of spinal root impairment and predicting prognosis in patients with lumbar spinal stenosis (LSS). The purpose of this study was to investigate the efficacy of intraoperative direct electrical stimulation of the spinal root and the measurement of DML in LSS. METHODS: In 39 patients with LSS, a total of 93 spinal roots were stimulated, and evoked electromyography was recorded at the leg muscles after decompression. DML was measured and its correlation with clinical severity, as evaluated by Zurich claudication questionnaire (ZCQ) and Short Form 36 (SF-36), was investigated. RESULTS: For the stimulation of the L3, L4, and L5 spinal root, the mean DML (ms) were 6.8 (±1.4), 7.4 (±1.3), and 6.0 (±1.3) in gluteus medius, 9.3 (±1.5), 9.2 (±1.5), and 9.0 (±1.6) in biceps femoris, 9.7 (±1.0), 9.8 (±1.8), and 9.4 (±1.2) in vastus medialis, 16.1 (±1.0), 14.7 (±1.3), and 14.1 (±1.5) in tibialis anterior, and 16.4 (±1.4), 14.3 (±1.8), and 13.9 (±1.9) in gastrocnemius muscles. Statistically significant positive correlations were observed between DML and height. Preoperative symptom and function scores of ZCQ and postoperative bodily pain scores of SF-36 were significantly worse in the patients with prolonged DML. CONCLUSIONS: DML is thought to be useful for estimating the severity of spinal root impairment and for predicting the prognosis.

Anti-Semaphorin 3A neutralization monoclonal antibody prevents sepsis development in lipopolysaccharide-treated mice.

Semaphorin 3A (Sema3A), originally identified as a potent growth cone collapsing factor in developing sensory neurons, is now recognized as a key player in immune, cardiovascular, bone metabolism and neurological systems. Here we established an anti-Sema3A monoclonal antibody that neutralizes the effects of Sema3A both in vitro and in vivo. The anti-Sema3A neutralization chick IgM antibodies were screened by combining an autonomously diversifying library selection system and an in vitro growth cone collapse assay. We further developed function-blocking chick-mouse chimeric and humanized anti-Sema3A antibodies. We found that our anti-Sema3A antibodies were effective for improving the survival rate in lipopolysaccharide-induced sepsis in mice. Our antibody is a potential therapeutic agent that may prevent the onset of or alleviate symptoms of human diseases associated with Sema3A.