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Starting from readily available tertiary alcohols, four different reactions (a 1,3-migration of a hydroxy group, kinetic resolution, racemization, and an intramolecular Diels–Alder reaction) took place under co-catalysis by lipase and oxovanadium compounds in a one-pot process to produce polysubstituted tricyclic carbon frameworks in high yields and with high enantioselectivities. The key to the success of this process was the discovery that a silyl group attached to the terminal carbon of the vinyl moiety completely controls the direction of hydroxy group migration

The semiclassical tunneling method is applied to evaluate the tunneling splitting of tropolone due to the intramolecular proton transfer in the electronic excited state, first time, in a framework of the trajectory on-the-fly molecular dynamics (TOF-MD) approach. To prevent unphysical zero-point vibrational energy transfer among the normal modes of vibration, quantum zero-point vibrational energies are assigned only to the vibrational modes related to intramolecular proton transfer, whereas the remaining modes are treated as bath modes. Practical ways to determine the tunnel-initiating points and tunneling path are introduced. It is shown that the tunneling splitting decreases as the bath-mode energy increases. The experimental tunneling splitting value is well reproduced by the present TOF-MD approach based on the Wentzel-Kramers-Brillouin (WKB) approximation.

SUMMARYThe ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has threatened human health and the global economy. Development of additional vaccines and therapeutics is urgently required, but such development with live virus must be conducted with biosafety level 3 confinement. Pseudotyped viruses have been widely adopted for studies of virus entry and pharmaceutical development to overcome this restriction. Here we describe a modified protocol to generate vesicular stomatitis virus (VSV) pseudotyped with SARS-CoV or SARS-CoV-2 Spike protein in high yield. We found that pseudovirions produced with the conventional transient expression system lacked coronavirus Spike protein at their surface as a result of inhibition of parental VSV infection by overexpression of this protein. Establishment of stable cell lines with an optimal expression level of coronavirus Spike protein allowed the efficient production of progeny pseudoviruses decorated with Spike protein. This improved VSV pseudovirus production method should facilitate studies of coronavirus entry and development of antiviral agents.

Endocytosis mediates the internalization and ingestion of a variety of endogenous or exogenous substances, including virus particles, under the control of intracellular signaling pathways. We have previously reported that the complex formed between the small GTPase Ras and phosphoinositide 3-kinase (PI3K) translocates from the plasma membrane to endosomes, signaling from which thereby regulates clathrin-independent endocytosis, endosome maturation, influenza virus internalization, and infection. However, the molecular mechanism by which the Ras-PI3K complex is recruited to endosomes remains unclear. Here, we have identified the amino acid sequence responsible for endosomal localization of the Ras-PI3K complex. PI3K lacking this sequence failed to translocate to endosomes, and expression of the peptide comprising this PI3K-derived sequence inhibited clathrin-independent endocytosis, influenza virus internalization, and infection. Moreover, treatment of cells with this peptide in an arginine-rich, cell-penetrating form successfully suppressed influenza virus infection in vitro and ex vivo, making this peptide a potential therapeutic agent against influenza virus infection.Key words: signal transduction, endocytosis, endosome, imaging, influenza virus.

SUMMARYIntracellular organelles of mammalian cells communicate with each other during various cellular processes. The functions and molecular mechanisms of such interorganelle association remain largely unclear, however. We here identified voltage-dependent anion channel 2 (VDAC2), a mitochondrial outer membrane protein, as a binding partner of phosphoinositide 3-kinase (PI3K), a regulator of clathrin-independent endocytosis downstream of the small GTPase Ras. VDAC2 was found to tether endosomes positive for the Ras-PI3K complex to mitochondria in response to cell stimulation with epidermal growth factor and to promote clathrin-independent endocytosis as well as endosome maturation at membrane contact sites. With a newly developed optogenetics system to induce mitochondrion-endosome association, we found that, in addition to its structural role in such association, the pore function of VDAC2 is also required for the promotion of endosome maturation. Our findings thus uncover a previously unappreciated role of mitochondrion-endosome association in the regulation of endocytosis and endosome maturation.HighlightsThe mitochondrial protein VDAC2 binds PI3K and tethers endosomes to mitochondriaVDAC2 promotes clathrin-independent endocytosisVDAC2-PI3K interaction induces acidification of endosomes associated with mitochondriaThe pore function of VDAC2 also contributes to endosome maturation at contact sites

To the EditorWe formulated mass spectral fragmentations of lipids across 117 lipid subclasses and included ion mobility tandem mass spectrometry (MS/MS) to provide a comprehensive lipidome atlas with retention time, collision cross section, and MS/MS information. The all-in-one solution from import of raw MS data to export of a common output format (mztab-M) was packaged in MS-DIAL 4 (http://prime.psc.riken.jp/) providing an enhanced standardized untargeted lipidomics procedure following lipidomics standards initiative (LSI) semi-quantitative definitions and shorthand notation system of lipid structures with a 1–2% estimated false discovery rate, which will contribute to harmonizing lipidomics data across laboratories to accelerate lipids research.

We present Mass Spectrometry-Data Independent Analysis software version 4 (MS-DIAL 4), a comprehensive lipidome atlas with retention time, collision cross-section and tandem mass spectrometry information. We formulated mass spectral fragmentations of lipids across 117 lipid subclasses and included ion mobility tandem mass spectrometry. Using human, murine, algal and plant biological samples, we annotated and semiquantified 8,051 lipids using MS-DIAL 4 with a 1–2% estimated false discovery rate. MS-DIAL 4 helps standardize lipidomics data and discover lipid pathways. Mass spectral fragmentations of lipids across 117 lipid subclasses are presented in a lipidome atlas.

The discovery of fluorescent proteins (FPs) has revolutionized cell biology. The fusion of targeting sequences to FPs enables the investigation of cellular organelles and their dynamics; however, occasionally, such fluorescent fusion proteins (FFPs) exhibit behavior different from that of the native proteins. Here, we constructed a color pallet comprising different organelle markers and found that FFPs targeted to the mitochondria were mislocalized when fused to certain types of FPs. Such FPs included several variants of Aequorea victoria green FP (aqGFP) and a monomeric variant of the red FP. Because the FFPs that are mislocalized include FPs with faster maturing or folding mutations, the increase in the maturation rate is likely to prevent their expected localization. Indeed, when we reintroduced amino acid substitutions so that the FP sequences were equivalent to that of wild-type aqGFP, FFP localization to the mitochondria was significantly enhanced. Moreover, similar amino acid substitutions improved the localization of mitochondria-targeted pHluorin, which is a pH-sensitive variant of GFP, and its capability to monitor pH changes in the mitochondrial matrix. Our findings demonstrate the importance of selecting FPs that maximize FFP function.

Mass spectrometry raw data repositories, including Metabolomics Workbench and MetaboLights, have contributed to increased transparency in metabolomics studies and the discovery of novel insights in biology by reanalysis with updated computational metabolomics tools. Herein, we reanalyzed the previously published lipidomics data from nine algal species, resulting in the annotation of 1437 lipids achieving a 40% increase in annotation compared to the previous results. Specifically, diacylglyceryl-carboxyhydroxy-methylcholine (DGCC) in Pavlova lutheri and Pleurochrysis carterae, glucuronosyldiacylglycerol (GlcADG) in Euglena gracilis, and P. carterae, phosphatidylmethanol (PMeOH) in E. gracilis, and several oxidized phospholipids (oxidized phosphatidylcholine, OxPC, phosphatidylethanolamine, OxPE, phosphatidylglycerol, OxPG, phosphatidylinositol, OxPI) in Chlorella variabilis were newly characterized with the enriched lipid spectral databases. Moreover, we integrated the data from untargeted and targeted analyses from data independent tandem mass spectrometry (DIA-MS/MS) acquisition, specifically the sequential window acquisition of all theoretical fragment-ion MS/MS (SWATH-MS/MS) spectra, to increase the lipidomic annotation coverage. After the creation of a global library of precursor and diagnostic ions of lipids by the MS-DIAL untargeted analysis, the co-eluted DIA-MS/MS spectra were resolved in MRMPROBS targeted analysis by tracing the specific product ions involved in acyl chain compositions. Our results indicated that the metabolite quantifications based on DIA-MS/MS chromatograms were somewhat inferior to the MS1-centric quantifications, while the annotation coverage outperformed those of the untargeted analysis of the data dependent and DIA-MS/MS data. Consequently, integrated analyses of untargeted and targeted approaches are necessary to extract the maximum amount of metabolome information, and our results showcase the value of data repositories for the discovery of novel insights in lipid biology.

Brown algae produce alginate that has various ratios and diverse sequences of two uronic acids, beta-D-mannuronic acid and alpha-L-guluronic acid, compared with those of alginate produced by bacteria. This diversity of alginate in brown algae is caused by mannuronan C5-epimerases (MC5Es), which catalyze the conversion of beta-D-mannuronic acid to alpha-L-guluronic acid. Although several bacterial MC5E enzymes have been well characterized, to date, there exists no information on the biochemical properties of eukaryotic MC5E. In this study, MC5E expression was detected in a brown alga Saccharina japonica sporophyte by immunoblot analysis. We also searched for MC5E mRNA from S. japonica by RT-PCR and revealed eight partial amino acid sequences, SjC5-I to -VIII. We focused on the highest frequency clone, SjC5-VI, and elucidated its full-length cDNA and putative gene structure. The translated SjC5-VI protein consists of 499 amino acids, with the N-terminal 21 amino acids predicted as a secretion signal sequence. Functional recombinant SjC5-VI (rSjC5-VI) was successfully expressed as a secreted protein using an insect-cell expression system, and we determined the optimal temperature, pH, and NaCl concentrations to be 35 degrees C, 7.0-8.2, and 300 mM, respectively, using the Ca2+-induced gel-formation assay. In addition, Ca2+ enhanced gelation by 1.7-fold following rSjC5-VI activity. Furthermore, H-1-NMR spectroscopy of rSjC5-VI-treated polyM revealed alternate epimerization of beta-D-mannuronic acid to alpha-L-guluronic acid. To the best of our knowledge, this is the first report on the characterization of MC5E activity in eukaryotes. (C) 2016 Elsevier B.V. All rights reserved.

Influenza A virus (IAV) infection is initiated by the attachment of the viral glycoprotein hemagglutinin (HA) to sialic acid on the host cell surface. However, the sialic acid-containing receptor crucial for IAV infection has remained unidentified. Here, we show that HA binds to the voltage-dependent Ca

Although the co-development of companion diagnostics with molecular targeted drugs is desirable, truly efficient diagnostics are limited to diseases in which chromosomal translocations or overt mutations are clearly correlated with drug efficacy. Moreover, even for such diseases, few methods are available to predict whether drug administration is effective for each individual patient whose disease is expected to respond to the drug(s). We have previously developed a biosensor based on the principle of Forster resonance energy transfer to measure the activity of the tyrosine kinase BCR-ABL and its response to drug treatment in patient-derived chronic myeloid leukemia cells. The biosensor harbors CrkL, one of the major substrates of BCR-ABL, and is therefore named Pickles after phosphorylation indicator of CrkL en substrate. The efficacy of this technique as a clinical test has been demonstrated, but the number of cells available for analysis is limited in a case-dependent manner, owing to the cleavage of the biosensor in patient-derived leukemia cells. Here, we describe an improved biosensor with an amino acid substitution and a nuclear export signal being introduced. Of the two predicted cleavage positions in CrkL, the mutations inhibited one cleavage completely and the other cleavage partially, thus collectively increasing the number of cells available for drug evaluation. This improved version of the biosensor holds promise in the future development of companion diagnostics to predict responses to tyrosine kinase inhibitors in patients with chronic myeloid leukemia.

Cellular interactions with the extracellular matrix play critical roles in tumor progression. We previously reported that receptor activator of NF-κB ligand (RANKL) specifically facilitates head and neck squamous cell carcinoma (HNSCC) progression in vivo. Here, we report a novel role for RANKL in the regulation of cell adhesion. Among the major type I collagen receptors, integrin α2 was significantly upregulated in RANKL-expressing cells and its knockdown suppressed cell adhesion. The mRNA abundance of integrin α2 positively correlated with that of RANKL in human HNSCC tissues. We also revealed that RANK-NF-κB signaling mediated integrin α2 expression in an autocrine/paracrine manner. Interestingly, the amount of active integrin β1 on the cell surface was increased in RANKL-expressing cells through the upregulation of integrin α2 and endocytosis. Moreover, the RANK-integrin α2 pathway contributed to RANKL-dependent enhanced survival in a collagen gel and inhibited apoptosis in a xenograft model, demonstrating an important role for RANKL-mediated cell adhesion in three-dimensional environments.

Influenza A virus (IAV) infection is initiated by the attachment of the viral glycoprotein hemagglutinin (HA) to sialic acid on the host cell surface. However, the sialic acid-containing receptor crucial for IAV infection has remained unidentified. Here, we show that HA binds to the voltage-dependent Ca2+ channel Cav1.2 to trigger intracellular Ca2+ oscillations and subsequent IAV entry and replication. IAV entry was inhibited by Ca2+ channel blockers (CCBs) or by knockdown of Cav1.2. The CCB diltiazem also inhibited virus replication in vivo. Reintroduction of wild-type but not the glycosylation-deficient mutants of Cav1.2 restored Ca2+ oscillations and virus infection in Cav1.2-depleted cells, demonstrating the significance of Cav1.2 sialylation. Taken together, we identify Cav1.2 as a sialylated host cell surface receptor that binds HA and is critical for IAV entry.

There have been several reports that photodesorption due to synchroton radiation from OFC copper chambers with machined surfaces is far lower than in stainless steel or aluminum alloy chambers. However, it is difficult to process machined surfaces for the inner surfaces of beam chambers. A surface treatment using broaching has been developed to cut the inner surface of the chamber. Tests of the photodesorption caused by synchrotron radiation in a small, broached, cylindrical cross-section chamber with a length of 30 cm have been reported. To adapt the broaching method to long chambers, prototype copper vacuum chambers with lengths of 1.8 m and 2 m have been fabricated. A chamber consists of a beam channel, a cooling channel, and a NEG pump channel. The beam channel, which has a hexagonal cross section, is broached. An ultimate pressure of 1 × 10 −8 Pa was obtained after bakeout at 200 °C for 24 h and NEG activation.

Angiotensin II (AII) type 2 receptor (AT2R) negatively regulates type 1 receptor (AT1R) signaling. However, the precise molecular mechanism of AT2R-mediated AT1R inhibition remains poorly understood. Here, we characterized the local and functional interaction of AT2R with AT1R. AT2R colocalized and formed a complex with AT1R at the plasma membrane, even in the absence of AII. Upon AII stimulation, the spatial arrangement of the complex was modulated, as confirmed by Förster resonance energy transfer (FRET) analysis, followed by AT2R internalization along with AT1R. AT2R internalization was specifically observed only in the presence of AT1R; AT2R alone could not be internalized. The AT1R-specific inhibitor losartan completely inhibited both the conformational change and the internalization of AT2R with AT1R, whereas the AT2R-specific inhibitor PD123319 partially hindered these phenomena, demonstrating that the activation of both receptors was indispensable for these effects. In addition, treatment with the protein kinase C (PKC) inhibitors inhibited the ligand-dependent accumulation of AT2R but not that of AT1R in the endosomes. A mutation in the putative phosphorylation sites of AT2R also abrogated the co-internalization of ATR2 with AT1R and the inhibitory effect of ATR2 on AT1R. These data suggest that AT2R inhibits ligand-induced AT1R signaling through the PKC-dependent pathway.

To study whether the suppression of the thyrotropin (thyroid-stimulating hormone, TSH) response to thyrotropin-releasing hormone (TRH) correlates with severity of illness and death in patients with nonthyroidal critical illness.Prospective study.Intensive care unit (ICU) of a university hospital.Forty-one critically ill patients without thyroid disease with multiple organ failure who were admitted to the ICU.The TSH response to TRH was tested within 24 hrs of ICU admission. Blood samples were obtained just before, and at 15, 30, 60, 90, and 120 mins after 500-micrograms injection of synthetic TRH. Triiodothyronine, free-triiodothyronine, thyroxine, free-thyroxine and TSH concentrations were measured in the samples obtained just before TRH injection. Acute Physiology and Chronic Health Evaluation (APACHE II) scores and Sepsis scores were calculated based on the data obtained within 24 hrs of ICU admission. Individual variables were compared between survivors and nonsurvivors. The APACHE II scores and Sepsis scores of nonsurvivors were significantly higher than those scores of survivors. The overall occurrence of suppressed TSH response to TRH was 88%. Peak TSH concentration of the TSH response was significantly lower in nonsurvivors than in survivors. Serial measurement of the TSH response showed that nonsurvivors experienced a decrease in peak TSH concentration from 1.55 +/- 0.78 to 0.55 +/- 0.30 microIU/mL; in survivors, it increased from 2.10 +/- 0.26 to 7.38 +/- 1.83 microIU/mL. Conversely, the basal TSH concentration did not change in either survivors or nonsurvivors. The "severity" of illness of nonsurvivors remained high; their mean APACHE II score varied from 20.0 +/- 1.9 to 22.1 +/- 1.3 and the mean Sepsis score varied from 20.0 +/- 4.3 to 25.4 +/- 4.0, while the same scores for survivors decreased significantly (p.05): their APACHE II score decreased from 16.2 +/- 0.7 to 7.6 +/- 2.0 and the Sepsis score went from 14.0 +/- 1.9 to 6.0 +/- 1.6.In critically ill patients with multiple organ failure, suppression of the TSH response to TRH frequently occurs and correlates with severity of illness and outcome. Our data indicate that measurement of the TSH response is helpful in evaluating the severity of illness and prognosis for critically ill patients.

Two controversial states considered to have a large alpha cluster strength, the 0{sup +} state at 8.54 MeV and the 1{sup {minus}} state at 11.7 MeV of {sup 44}Ti, were examined in the ({sup 6}Li,{ital d}) reaction at {ital E}({sup 6}Li)=50 MeV. The 11.7-MeV state was scarcely observed, if at all, for {theta}{sub lab}=5--35{degree}, and the angular distribution for the 8.54-MeV state was unlike that of an {ital L}=0 transition. These results indicate that both states are unsuitable candidates for the lowest states of the parity doublet band of {sup 44}Ti in the alpha cluster model.

A 57-year-old female patient underwent combined off-pump coronary artery bypass grafting and abdominal aortic aneurysm replacement. Anesthesia was maintained with propofol, fentanyl, and thoracic epidural anesthesia. Propofol doses were adjusted to maintain bispectral index (BIS) between 40-60. Despite the remarkable hemodynamic changes, BIS remained stable at about 50 during the surgery. The average dose of propofol was 3.3 mg.kg-1.hr-1. The patient awoke an hour after the surgery and was extubated 1.5 hours thereafter. This case report suggests that BIS is a useful index to determine the depth of anesthesia during surgeries which induce marked hemodynamic changes.

We investigated the effect of premixing lidocaine with propofol on a bispectral index (BIS) during propofol infusion. We studied 40 adult patients given mixture of 1% propofol 20 ml with 2 ml of normal saline (control group) or 2% lidocaine (lidocaine group) infused at 2 ml.kg-1.hr-1 for 10 minutes. Heart rate, mean arterial pressure, and BIS were measured every minute. The addition of lidocaine to propofol reduced the incidence of injection pain from 85% to 10% but did not change the induction time. Propofol significantly decreased mean arterial pressure and BIS but there was no difference between the groups. In conclusion, premixing lidocaine with propofol reduces injection pain without affecting the hypnotic effect.

We describe a case presenting herpetic eruptions and pain in the left cranial and cervical regions, followed by right cranial polyneuropathy. The patient, a 66-yr-old male, developed a diphasic syndrome which was first characterized by herpetic eruptions on the whole of his left cranial and cervical regions with severe pain and left facial nerve palsy. Two weeks later, right multiple cranial nerve palsies of III, V, and VII and an increase in CSF protein appeared in a second phase. Neuroimaging technique (MRI) did not provide any evidence of brain involvement. The immediate use of steroid therapy led to improvement of the symptoms of cranial polyneuropathy. The clinical course suggests that reactivation of a latent herpes simplex virus may have been the etiologic cause of the cranial polyneuropathy. Steroid therapy should be applied to the treatment of cranial polyneuropathy in patients showing a similar pattern to this case.

We investigated changes in plasma concentration of lidocaine during continuous epidural anesthesia in 21 patients undergoing hepatectomy. According to the extent of hepatectomy, patients were assigned into one of the three groups: partial resection group, lobectomy group, and extended lobectomy group. Lidocaine 2.0 mg.kg-1 with 1: 200,000 epinephrine was epidurally administered in a bolus followed by continuous infusion of lidocaine at the rate of 1.5mg.kg-1.h-1. Plasma lidocaine concentration in the extended lobectomy group showed a significantly higher value than those of other two groups (P0.05). Plasma lidocaine was detected in the extended lobectomy group at 12 hs after the surgery, but not in the other two groups. Furthermore we investigated the effects of prostaglandin E1 infusion in 7 patients undergoing extended lobectomy. Plasma lidocaine concentration was not increased in this group. In conclusion, we recommend caution regarding the dose of lidocaine administered for epidural anesthesia during hepatectomy; PGE1 infusion appears to be safe in patients undergoing extended hepatectomy.

This study was conducted to evaluate the effects of oral clonidine and tizanidine, alpha 2 adrenergic agonists, as premedication for tetracaine spinal anesthesia in 63 gynecological patients. The patients were randomly allocated to one of six groups. Group 1 (n = 7), group 2 (n = 8) and group 3 (n = 7) received 13 mg of tetracaine intrathecally in 10 % glucose solution 2.6 ml. Group 4 (n = 13), group 5 (n = 14) and group 6 (n = 13) received 13 mg of tetracaine intrathecally in a volume of 2.6 ml of 10 % glucose solution which contained 0.65 mg of phenylephrine. As premedication, group 1 and 4 received 0.25 mg of oral triazolam; group 2 and 5 received 3 mg of oral tizanidine; group 3 and 6 received 0.15 mg of oral clonidine. Group 2 and 3, or group 5 and 6 (clonidine or tizanidine group, respectively) needed significant longer time for regression of Th10 sensory blockade than group 1 or 4 (triazolam). The time for appearance of postoperative pain and the time to require postoperative analgesics were longer in the groups which had received either clonidine or tizanidine than in the groups which had received triazolam. Heart rate and systolic blood pressure in group 6 (clonidine-tetracaine-phenylephrine group) showed significant decreases (P0.05) after the spinal anesthesia. We concluded that oral premedication of clonidine and tizanidine prolonged tetracaine spinal anesthesia. From the view point of the prolongation of spinal anesthesia and the hemodynamic stability, oral premedication with tizanidine seems to be useful.

Buprenorphine suppository (BSP) has been available as an analgesic agent in Japan since 1990. It has gained considerable popularity in various clinical situations; however, its usefulness for postoperative pain relief has been controversial. The present study is addressed to postoperative pain in fifty-seven women who underwent transvaginal hysterectomy under spinal anesthesia with tetracaine and phenylephrine. We examined the analgesic effects of buprenorphine (0.4 mg) suppository. Patients were divided into four groups according to the timing of buprenorphine suppository administration: group A1, preoperative administration of BSP, n = 12; group A2, postoperative administration of BSP, n = 17; group A3, intravenous injection of 0.1 mg of buprenorphine during surgery and postoperative administration of BSP, n = 14; group C, control group, n = 14. All the patients in group C complained of pain within 24 hr. The onset time of pain after surgery was significantly longer in patients in group A1 (372 +/- 220 min) and A3 (481 +/- 161 min) than in control group C (282 +/- 97 min). The percentage of patients who did not complain of pain within 24 hr was the largest in group A3 (36%). However there were no significant differences in age, body weight, effective analgesic time, analgesic level or operation time among the four groups. Nausea and vomiting were observed in all groups: C (36%), A1 (41%), A2 (18%), and A3 (29%). From these results, we conclude that preoperative or intraoperative administration of buprenorphine is useful for control of postoperative pain; the method of administration of buprenorphine suppository needs further study.

A 57-yr-old male with atrial septal defect (ASD) was scheduled for the patch closure operation. The patient had a history of hyperthyroidism due to giant adenoma of the thyroid gland. The patient was controlled under euthyroid state by thiamazole for four years. With this treatment, his thyroid function became normal and he was doing well for over the last seven years. On the morning of the day of operation, thiamazole was given orally to this patient. When the ASD patch closure was performed, the examination of his thyroid gland revealed hypothyroidism, but the operation could be performed without any thyroidal trouble. After the operation thiamazole was given intramuscularly to this patient and from the next day it was continued orally. Thyroid storm did not occur after the operation until discharge. We conclude that in a case of heart disease with hyperthyroidism, it is important to keep the patient's thyroid function under normal for a long time before surgery.

We performed transcatheter hepatic segmental chemo-Lipiodol-embolization against hepatocellular carcinoma accompanied by tumor thrombosis in the portal vein. CT scan after embolization demonstrated good Lipiodol accumulation in the tumor thrombosis in some cases. In one resected case, remarkable necrosis of tumor thrombosis was proved histologically. In five non-resected cases, 6-month survival rate was 100%. We hope this therapeutic method can be useful for hepatocellular carcinoma with tumor thrombosis in the portal vein.

A 71-year-old woman was transferred to Kushiro City General Hospital because of fever, sore throat, diffuse neck swelling and dyspnea. She had received right mastectomy for breast cancer under general anesthesia 6 days before the admission. The lateral X-ray film of the neck revealed abscess in the retropharyngeal space and the retroesophageal space. CT scan revealed mediastinitis. Next day she received neck dissection for drainage of the abscess under general anesthesia. Although the posterior pharyngeal wall was swollen, endo-tracheal intubation was not difficult. Brown tinged and purplish pus was aspirated from the interspace of carotid sheath and trachea, the retropharyngeal space, and the superior mediastinal space. The infected site was irrigated with a lot of peroxide and saline and draining tubes were placed in each interspace. Tracheostomy was not done but the patient was admitted to the ICU with her trachea intubated. The day after operation, she was extubated. Three days after the operation chest X-ray revealed pyothorax and chest tube was inserted for drainage. Seven days after the operation she was transferred to the ENT ward. Thereafter her recovery course was uneventful. It seems that the deep neck infection was probably caused by the injury on endotracheal intubation at the first operation in this case. Although this patient was cured of mediastinitis following deep neck infection, which is still lethal, early diagnosis and surgical drainage of the abscess are necessary.

We studied the effect of a low-dose intrathecal morphine (0.1 or 0.2 mg) in postoperative pain relief and the incidence of side effects. Two hundred and fifteen patients scheduled for transvaginal hysterectomy were divided into 3 groups according to intrathecal morphine doses: M1 (morphine 0.1 mg N = 75), M2 (morphine 0.2 mg N = 69) and C (control N = 71). A standard mid-line lumbar puncture was performed using a 25-gauze needle in the L3/4 interspace. Preservative-free morphine hydrochloride mixed in hyperbaric tetracaine solution was administered intrathecally. Pain relief was significantly greater for the first 24 hrs in groups M1 and M2 compared with group C. Respiratory depression was not seen in any groups. The incidence of vomiting was about 40% in all groups. We conclude that intrathecal morphine 0.1-0.2 mg is useful for pain relief after transvaginal hysterectomy and accompanies no major side effects.

We acquired dynamic images over the whole liver by inversion recovery snap shot FLASH method after a bolus intravenous injection of Gadolinium-DTPA. Each nodule of hepatoma in the liver showed early enhancement and gradually turned to show low intensity. In two cases of hepatoma, small intrahepatic metastases, which were not detected by US, CT and spin-echo image of MRI, were suspected as high intensity nodules on early phase. Also recurrent areas after TAE were enhanced on early phase. This method is practical for improving the detection of lesions and is useful for evaluating the recurrence after TAE.

The practical limitations of catheters used for infusion were studied by their types in 47 patients with hepatocellular carcinoma who were treated by Segmental Lp-TAE. For catheterization into the arterial branches of S5 and S8, a coaxial infusion catheter was helpful. However, a coaxial infusion catheter was seldom used for catheterization into the arterial branches of S6 and S7. For infusion into vessels having diameters of less than 1.9 mm at the tip end of the catheter, positive use of a coaxial infusion catheter is strongly recommendable. A Tracker-18 catheter was especially helpful for catheterization into the vessels of S5.

The evaluation of TAE was studied for 73 patients with metastatic liver cancers but without metastasis in other organs, who were treated with initial TAE more than 2 years ago. Seventeen patients survived for more than 2 years. The primary lesions consisted of colon cancer in 9 patients, the Langerhans islet cell tumor of the pancreas in 4, gastric cancers in 2, breast cancer in 1 and leiomyosarcoma of the colon in 1. The overall 2-year survival rate calculated by the direct method was 23.3% and mean survival period was 15.8 months. The prognosis was good in H1 and V3. Lp-TAE produced better effects but no statistically significant difference was found between the 2-year survivals by Lp-TAE and by GS-TAE.

The (d, d s =0 ) reaction has been studied for the first time at E d = 56 MeV and 70 MeV. It is demonstrated that the reaction is a useful tool for the study of the inelastic excitation of the spin- and isospin-flip mode. Several states in 12 C with T = 1 and S = 1 are strongly excited. The differential cross sections and vector analyzing powers are reasonably reproduced by the distorted-wave Born-approximation calculation.

A precision gas feeding system aiming at stable and efficient operation of a small, cold-cathode PIG heavy ion source for the K = 50 MeV AVF cyclotron is described in detail. The system can regulate the supply gas pressure with a precision of 0.6 Torr and keep a constant gas flow rate with a stability of 0.010 ± 0.003 cm3/min. The ion source operating parameters such as the arc current, the arc voltage and the gas flow rate remain fairly constant through the lifetime of the source. By adopting this system in the routine accelerations of carbon or oxygen ions, the average beam current was increased by a factor of 2–3.

The ($\ensuremath{\alpha},{d}_{S=0}$) reaction was studied for the first time with a $^{12}\mathrm{C}$ target at ${E}_{\ensuremath{\alpha}}=50$ MeV. Three $T=1$ states in $^{14}\mathrm{N}$, ${0}^{+}$ at 2.31 MeV, ${3}^{\ensuremath{-}}$ at 8.91 MeV, and a level at 12.6 MeV, were selectively excited. The experimental results were well described by exact finite-range distorted-wave calculations.

An on-line multi-data acquisition controller system has been designed to be connected to a DRU-11 interface as a peripheral to a PDP11/44 computer (a host). The system was specifically designed for practical use of multiple detector systems which consist of two ΔE detectors and a position sensitive detector.

Although cAMP is well established as a second messenger for olfactory transduction in vertebrates, the role of inositol 1,4,5-trisphosphate (IP3) in this process remains controversial. We addressed this issue by comparing currents evoked by cAMP and IP3 in native and reconstituted membranes from olfactory cilia. We detected only a cyclic nucleotide-gated conductance in the native membrane but both cyclic nucleotide-gated and IP3-gated conductances in the reconstituted membrane. The magnitudes of the cyclic nucleotide- and IP3-gated conductances were not correlated with each other in reconstituted membranes, suggesting that cyclic nucleotide- and IP3-gated channels originate in different cellular compartments.

The patients with gallbladder cancer often accompany with chronic cholecystitis. This fact leads angiographic diagnosis difficult. So the angiographic findings of the chronic cholecystitis are first analyzed precisely and subtracting the angiographic findings of the gallbladder cancer from these gives us the pure (true) angiographic findings of the gallbladder cancer. Characteristic angiographic findings of the gallbladder cancer only are stretched cystic artery encasement (27/37 cases) and short straight tumor vessels "bristly vessel" (36/37 cases).

Evaluation of "Gianturco-Wallace chemotherapy pulser," which was developed to produce a more homogeneous drug distribution of the tumors in intra-arterial infusion chemotherapy, was assessed by comparative study of pulsed and nonpulsed arterial radionuclide infusion using Tc-99m pertechnetate for 18 cases of hepatic carcinomas (11 cases of hepatocellular carcinomas and 7 cases of metastatic hepatic carcinomas). Tc-99m pertechnetate, 740 MBq (20 mCi) diluted with saline (30 mL) was infused with or without pulse through the catheter into the hepatic artery at a rate of 1mL per minute. The intrahepatic dynamic radionuclide distribution was analyzed by the time activity curves of ROIs in the tumor and nontumor areas. Pulsed infusion interrupted laminar flow and produced more homogeneous radionuclide distribution in the liver, and combination of pulsed and nonpulsed infusion also produced better radionuclide distribution in the areas of the tumors. This method using Tc-99m pertechnetate was very useful as a simulation to determine the dynamic drug distribution of the tumor and non-tumor region in intraarterial infusion methods.