Histological analysis of nucleus pulposus tissue from patients with lumbar disc herniation after condoliase administration.

Background: Condoliase is an enzyme used as a treatment for lumbar disc herniation (LDH). This enzyme degrades chondroitin sulfate (CS) in the nucleus pulposus of the intervertebral disc (IVD). However, there are cases in which symptoms do not improve, despite condoliase administration. This study reports histological analysis of lumbar disc tissue of LDH patients who underwent surgery because condoliase had no therapeutic effect. Methods: Between March 2019 and August 2019, 12 LDH patients who underwent full endoscopic spine surgery (FESS) discectomy at the Dezawa Akira PED Clinic were the subjects of the study. There are two study groups: six cases underwent FESS after condoliase administration, while six underwent FESS without condoliase administration. The average duration from drug administration to surgery was 152 days. Herniated disc removed at surgery was evaluated by histological staining including immunohistochemistry by anti-CS antibodies. Results: Multiple large clusters (40-120 µm in diameter) were observed in the nucleus pulposus of those who received condoliase, but no clusters were observed in those who did not. The lumbar disc tissues, including the nucleus pulposus of recipients, were stained with anti-CS antibodies that recognize the CS unsaturated disaccharide, but non-administration tissue was not stained. These findings suggest that the enzyme acted on the nucleus pulposus, even in cases where symptoms were not improved by condoliase administration. Furthermore, there was no histological difference between stained images of the extracellular matrix in those who did or did not receive condoliase, suggesting that condoliase acted specifically on CS in the nucleus pulposus. Conclusions: We demonstrated that CS in the nucleus pulposus was degraded in patients in whom condoliase did not have a therapeutic effect. Moreover, condoliase acts in human IVD without causing necrosis of chondrocytes and surrounding tissues.

Development of in situ crosslinked hyaluronan as an adjunct to vitrectomy surgery.

Ophthalmologists have used hyaluronan (HA) products as adjuncts to ocular surgery since the 1970s. However, HA products are not always functional in surgeries of the posterior eye segment due to their lack of biomechanical strength. In this study, we developed an in situ crosslinked HA (XL-HA) and evaluated its potential as an adjunct to vitrectomy surgery in an in vitro model with a triamcinolone acetonide (TA) layer used as a pseudo residual vitreous cortex (RVC). Within a few minutes at concentrations over 0.9%, XL-HA, generated by the click chemistry of HA-dibenzocyclooctyne and HA-azidoethylamine, formed a hydrogel with the appropriate hardness for tweezers peeling. XL-HA (concentration, 0.76-1.73%) without dispersion successfully entered the TA layer and removed more than 45% of the total TA. Dynamic viscoelasticity helps to explain the rheological behavior of hydrogels, and the assessment results for XL-HA indicated that suitable concentrations were between 0.97% and 1.30%. For example, 1.30% XL-HA hydrogel reached sufficient hardness at 3 min for tweezers peeling, and the TA removal ability exceeded 70%. These results demonstrated that XL-HA was a potential adjunct to successful vitrectomy.

[Clinical Functionality of Dispersive OVDs: Improvement of One of the Properties of 3% Hyaluronic Acid and 4% Chondroitin Sulfate Combination].

Ophthalmic viscosurgical devices (OVDs) are mainly divided into two general categories: cohesive and dispersive. Dispersive OVDs such as the 3% hyaluronic acid and 4% chondroitin sulfate (HA/CS) combination have excellent adhesion to ocular tissues and protect the corneal endothelium to a greater extent than cohesive OVDs. Herein, we summarize our recent findings regarding one of the properties of the HA/CS combination related to clinical performance. (i) The room temperature stability of OVDs and needle clogging by OVDs remain clinical issues. We demonstrated that adding d-sorbitol to the HA/CS combination preserved its viscosity, which was equivalent after 2 year-storage at room temperature to the viscosity of HA/CS combination stored under refrigeration for 2 years without d-sorbitol. Besides, the HA/CS combination with d-sorbitol could be used repeatedly without cleaning or replacing the needle, suggesting that the addition of d-sorbitol prevents drying and solidification of the OVD on the needle. (ii) Although it can be inferred from numerous studies that the tissue adhesion of OVDs influences their retention by the eye, little is known about the physical properties of OVDs that contribute to intraocular retention. To address this issue, we compared two types of adhesive forces, detachment force and repulsive force, for each OVD. Compared with other dispersive OVDs, the HA/CS combination showed higher values for both adhesive forces. These results suggest that adhesive forces may be used as an index of dispersive OVD retention in the eye.

Cardiac macrophages regulate isoproterenol-induced Takotsubo-like cardiomyopathy.

Takotsubo syndrome (TTS) is an acute, stress-induced cardiomyopathy that occurs predominantly in women after extreme physical and/or emotional stress. To date, our understanding of the molecular basis for TTS remains unknown and, consequently, specific therapies are lacking. Myocardial infiltration of monocytes and macrophages in TTS has been documented in clinical studies. However, the functional importance of these findings remains poorly understood. Here, we show that a single high dose of isoproterenol (ISO) in mice induced a TTS-like cardiomyopathy phenotype characterized by female predominance, severe cardiac dysfunction, and robust myocardial infiltration of macrophages. Single-cell RNA-Seq studies of myocardial immune cells revealed that TTS-like cardiomyopathy is associated with complex activation of innate and adaptive immune cells in the heart, and macrophages were identified as the dominant immune cells. Global macrophage depletion (via clodronate liposome administration) or blockade of macrophage infiltration (via a CCR2 antagonist or in CCR2-KO mice) resulted in recovery of cardiac dysfunction in ISO-challenged mice. In addition, damping myeloid cell activation by HIF1α deficiency or exposure to the immunomodulatory agent bortezomib ameliorated ISO-induced cardiac dysfunction. Collectively, our findings identify macrophages as a critical regulator of TTS pathogenesis that can be targeted for therapeutic gain.

Advances in Understanding the Mechanism of Ophthalmic Viscosurgical Device Retention in the Anterior Chamber or on the Corneal Surface during Ocular Surgery.

Retention durability, especially in the eye, is one of the most important properties of ophthalmic viscosurgical devices (OVDs) during ocular surgery. However, the information on the physical properties of OVDs is insufficient to explain their retention durability. The purpose of this study is to clarify the mechanism of OVD retention to improve understanding of the behavior of OVDs during ocular surgery. To elucidate the mechanism of OVD retention, we have developed a new test method for measuring repulsive force. As a result, the maximum repulsive force of OVDs was positively and well correlated with the retention durability of investigated OVDs. Consequently, we demonstrated that the repulsive force could be used as an index of retention durability on the ocular surface and in the eye. We directly compared the intraocular retention durability of three OVDs (Shellgan, Viscoat, and Opegan-Hi) in ex vivo porcine eyes. Opegan-Hi was immediately removed from the anterior chamber, but Shellgan and Viscoat remained largely in the anterior chamber as determined by fluorescence imaging. These results showed that the intraocular retention behavior of OVDs was similar to their ocular surface behavior in our previous report, suggesting that retention durability is dependent on the OVD itself. The retention durability of Shellgan seemed to be higher than that of Viscoat, and the maximum repulsive force of Shellgan was 1.35-fold higher than that of Viscoat. Therefore, the repulsive force might be a useful index for assessing the difference in the retention durability between OVDs such as Shellgan and Viscoat.

Quantitative Assessment of Ophthalmic Viscosurgical Devices on Visibility, Spreadability, and Durability as Corneal Wetting Agents for the Wet Shell Technique.

PURPOSE: To evaluate ophthalmic viscosurgical devices (OVDs) as corneal wetting agents for the wet shell technique, a common procedure in Japan to maintain the wettability of corneal surfaces. METHODS: We surveyed Japanese ophthalmologists to determine the current state of the wet shell technique. After developing three ex vivo testing methods, we evaluated the corneal wetting properties of OVDs including 3% hyaluronic acid (HA) solution and OVD products, Opegan, Opelead, Viscoat, Shellgan, Discovisc, and Opegan-Hi. RESULTS: Overall, 214 ophthalmologists (70%) had performed the wet shell technique, and 91% of ophthalmologists who performed vitreous surgery had performed this technique. Using a questionnaire, we evaluated the performance of OVD as corneal wetting agents as follows: (i) visibility, smoothness of OVD surface; (ii) spreadability, coverage of the cornea; and (iii) retention durability, residual ratio of OVD on the corneal surface. The smoothness and spreadability of Opegan, Opelead, and 3% HA were superior to other OVDs. Adding an appropriate amount of balanced salt solution to the other OVDs improved smoothness and spreadability similar to that of Opelead or 3% HA. Shellgan and Viscoat, combination OVDs consisting of 3% HA and 4% chondroitin sulfate, showed high retention durability, resulting in remaining longer on the cornea compared with other OVDs. CONCLUSIONS: Physical properties of OVDs tested in this study may provide useful information for ophthalmologists to select a suitable OVD when performing the wet shell technique.

D-sorbitol can keep the viscosity of dispersive ophthalmic viscosurgical device at room temperature for long term.

The combination of 3% sodium hyaluronate (HA) and 4% sodium chondroitin sulfate (CS) is used as a dispersive ophthalmic viscosurgical device (OVD) during cataract surgery. For most OVDs containing HA, storage at 2-8 °C is recommended to preserve product characteristics. In order to develop a dispersive OVD that can be stored at room temperature, in this study, we searched additives which can stably maintain the viscosity, a key parameter of OVD, under preservation stability testing at 60 °C. The addition of D-sorbitol to a combination OVD, 3% HA and 4% CS, suppressed the reduction in viscosity compared with other OVDs with or without additives. The addition of D-sorbitol was also effective in improving the residual viscosity of the combination OVD after thermal treatment and light irradiation. Moreover, the OVD containing D-sorbitol can be stored at 25 °C with stably maintaining the initial viscosity for at least 24 months. In conclusion, the new dispersive OVD, 3% HA, 4% CS, and 0.5% D-sorbitol, can be stored at room temperature instead of under cold conditions and may represent an attractive option for clinical use because it is not necessary to bring the product to room temperature prior to use.

Addition Of D-Sorbitol Improves The Usability Of Ophthalmic Viscosurgical Devices.

PURPOSE: To evaluate the effects of D-sorbitol addition on changes in the extrusion force of ophthalmic viscosurgical devices (OVDs). METHODS: OVD formulations; the mixtures of 3% hyaluronic acid (HA) and 4% chondroitin sulfate (CS) containing 0%, 0.5%, or 1.0% D-sorbitol were prepared. Each prefilled syringe of OVD was stored at room temperature for 0, 15, 30, 60, or 120 mins after a small amount of viscoelastic agent was discharged from the needle. The extrusion force values (kgf) of these OVDs when reused after storage were measured with a texture analyzer. Moreover, 10 healthy adults (5 men and 5 women) used a pinch sensor to measure the extrusion force values for the HA/CS combination without D-sorbitol which was stored in the above manner, and used a 4-step scale to score the usability of OVD. RESULTS: For the HA/CS combination without D-sorbitol, the extrusion force value was increased from its initial value (storage duration, 0 min) as storage duration increased. However, for the HA/CS combination containing 0.5% or 1.0% D-sorbitol, this value remained almost unchanged over time. Likewise, the pinch sensor-determined extrusion force values of HA/CS combination without D-sorbitol increased, depending on storage duration. CONCLUSION: The addition of D-sorbitol to viscoelastic agent may suppress the needle clogging that occurs with OVD storage, and may improve the usability of OVDs during surgery.

[Evaluation of Rheological Properties of Cohesive Ophthalmic Viscosurgical Devices Composed of Sodium Hyaluronate with High Molecular Weight-2019].

Ophthalmic viscosurgical devices (OVDs), mainly containing sodium hyaluronate (HA), are used in cataract surgeries to protect the cornea endothelium. In this study, the rheological properties of 9 launched products (containing 1% HA) were evaluated. The molecular weights (MWs) of HA estimated based on intrinsic viscosity varied widely, between 1100-2500 kDa, and showed a particular value for each product. Of the 9 products, 6 are classified as cohesive OVDs and their product specifications show the same value for intrinsic viscosity (25-45 dL/g), with high MW HA (>2000 kDa); however, the MW of each HA showed a particular value (2200-2500 kDa) within the range of the product specification. As with the MW of HA, apparent viscosity and dynamic rheological parameters showed particular values for each OVD. The product Opegan-Hi exhibited the highest value of apparent viscosity at low shear rate, and a solid-like behavior among the OVDs. In a questionnaire survey among 198 cataract surgeons, 42% of surgeons had experienced a difference in ability to maintain the depth of anterior chamber during surgery among the different cohesive OVDs used. This suggested that surgeons select OVD properties based on surgical procedure and patient cases. In conclusion, we demonstrated that each OVD has particular rheological properties within the range of the product specification defined by the intrinsic viscosity. The results might provide useful information for surgeons in their selection of OVDs based on their experience.

Rheological and Adhesive Properties to Identify Cohesive and Dispersive Ophthalmic Viscosurgical Devices.

The purpose of this study is to evaluate the usefulness of the rheological properties and adhesive force of ophthalmic viscosurgical devices (OVDs) as parameters for understanding and identifying the surgical behavior of cohesive and dispersive OVDs. The apparent viscosity, and the storage and loss moduli (dynamic rheological parameters) of 50% chondroitin sulfate (CS), 3% sodium hyaluronate (HA), Shellgan (the combination of 3% HA and 4% CS), Opegan (1% HA with a low molecular mass) and Opegan-Hi (1% HA with a high molecular mass) were obtained with a rheometer. The adhesive force of each sample was measured by using a texture analyzer. Opegan-Hi showed a solid-like behavior, while 50% CS showed a fluid-like behavior from their apparent viscosity and dynamic rheological parameters. Shellgan, 3% HA, and Opegan exhibited similar rheological properties and intermediate characteristics between Opegan-Hi and 50% CS, although their respective values were slightly different. Among these OVD samples, the adhesive force was higher in the order of 50% CS > Shellgan, 3% HA > Opegan > Opegan-Hi. The adhesive force of dispersive OVDs tended to be higher than that of cohesive OVDs, which correlated well with the removal times of OVDs from the eye that have previously been reported. In conclusion, we demonstrated that cohesive OVDs and dispersive OVDs have particular rheological and adhesive properties that can be applied to identify both types. These parameters obtained in this study provide useful information for a greater understanding and prediction of the behavior of OVDs in the eye during surgery.

Pericoronary adipose tissue ratio is a stronger associated factor of plaque vulnerability than epicardial adipose tissue on coronary computed tomography angiography.

This study was designed to clarify the influence of pericoronary adipose tissue (PAT) on plaque vulnerability using coronary computed tomography angiography (CCTA). A total of 103 consecutive patients who underwent CCTA and subsequent percutaneous coronary intervention (PCI) using intravascular ultrasound (IVUS) for coronary artery disease were enrolled. The PAT ratio was calculated as the sum of the perpendicular thickness of the visceral layer between the coronary artery and the pericardium, or the coronary artery and the surface of the heart at the PCI site, divided by the PAT thickness without a plaque in the same vessel. PAT ratios were divided into low, mid and high tertile groups. Epicardial adipose tissue (EAT) thickness was measured at the eight points surrounding the heart. Multivariate logistic analysis was performed to determine whether the PAT ratio is predictive of vulnerable plaques (positive remodeling, low attenuation and/or spotty calcification) on CCTA or echo-attenuated plaque on IVUS. The Hounsfield unit of obstructive plaques >50% was lower in the high PAT group than in the mid and low PAT groups (47.5 ± 28.8 vs. 53.1 ± 29.7 vs. 64.7 ± 27.0, p = 0.04). In multivariate logistic analysis, a high PAT ratio was an independent, associated factor of vulnerable plaques on CCTA (OR: 3.55, 95% CI: 1.20-10.49), whereas mean EAT thickness was not (OR: 1.22, 95% CI: 0.82-1.83). We observed a similar result in predicting echo-attenuated plaque on IVUS. PAT ratio on CCTA was an associated factor of vulnerable plaques, while EAT was not. These results support the important concept of local effects of cardiac adipose tissue on plaque vulnerability.

Isolation and characterization of monoclonal antibodies specific for chondroitin sulfate E.

Chondroitin sulfate E (CSE) is a polysaccharide containing mainly disaccharide units of D-glucuronic acid (GlcA) and 4,6-O-disulfated N-acetyl-D-galactosamine (GalNAc) residues (E-unit) in the amount of ∼ 60%. CSE is involved in many biological and pathological processes. In this study, we established new monoclonal antibodies, termed E-12C and E-18H, by using CSE that contained more than 70% of E-units as an immunogen. These antibodies recognized CSE but not other CSs isomers or dermatan sulfate (DS). We evaluated the reactivities of the antibodies to 6-O-sulfated CSA (6S-CSA) and DS (6S-DS) that possessed ∼ 60% of GalNAc (4S, 6S) moieties in their structures. Neither of the antibodies reacted with 6S-DS. The antibodies strictly distinguished the structural difference of GlcA and L-iduronic acid in the polysaccharide. Binding affinities of the antibodies were determined by a surface plasmon resonance assay using CSE and 6S-CSA. The binding affinities were strongly associated with the molecular weight of CSE and the E-unit content of 6S-CSA. Moreover, we demonstrated that the antibodies are applicable to histochemical analysis. In conclusion, the new anti-CSE monoclonal antibodies specifically recognize the E-unit of CSE. The antibodies will become useful tools for the investigation of the biological and pathological significance of CSE.

[Efficacy of repeated subcostal transversus abdominis plane blocks with 0.2% lidocaine via 18-gauge intravenous catheters for patients undergoing abdominal aortic aneurism surgery].

BACKGROUND: There is an increasing number of patients scheduled for abdominal aortic aneurysm resection in whom epidural anesthesia cannot be performed because of concomitant antiplatelet/anticoagulant therapy. Instead of epidural anesthesia for postoperative analgesia in such patients it is possible to use repeated bilateral subcostal transversus abdominis plane (TAP) blocks. METHODS: Four patients receiving antiplatelet/anticoagulant therapy for abdominal aortic aneurysm resection under general anesthesia were studied. After the completion of surgery and before emergence from anesthesia 18-gauge intravenous catheters were inserted bilaterally into subcostal TAP and 100 ml (50 ml on each side) of 0.2% lidocaine with 1/500,000 epinephrine were injected via the catheters twice daily until the second postoperative day. Pain intensity was assessed using a 0-10 numerical rating scale at rest and during movement, before and after each block. RESULTS: Numerical pain ratings at rest and during movement decreased after each block, and good analgesia was obtained. No complications such as nausea, vomiting or infection were observed in the postoperative period. CONCLUSIONS: These findings suggest that repeated bilateral subcostal TAP blocks with 0.2% lidocaine performed via 18-gauge intravenous catheters provide good postoperative analgesia after abdominal aortic aneurysm resection.

[Safety and beneficial effects of spinal morphine on the postoperative course of elderly patients undergoing surgical fixation of the femoral neck fracture].

BACKGROUND: We performed a retrospective study of the efficacy and safety of spinal anesthesia with 0.1 mg morphine in the postoperative course of elderly patients with femoral neck fracture. METHODS: Sixty patients with ages averaging 84 years participated in this study. Surgery was performed under spinal anesthesia. Patients were assigned to either a group receiving of 0.1 mg morphine added to isobaric bupivacaine (Group M) or a group receiving of isobaric bupivacaine alone (Group B). The frequency of analgesic use and the occurrence of adverse side effects during the first 48 hours after surgery were compared between the two groups. RESULTS: In the first 24 hours, the patients in Group M needed significantly less analgesics compared to Group B. The incidence of adverse side effects did not differ significantly between the groups, although nausea had a tendency to increase in Group M. One patient in Group M showed a mild decrease in oxygen saturation. CONCLUSIONS: The spinal administration of 0.1 mg morphine had beneficial effects and was safe in the postoperative period of elderly patients with femoral neck fracture provided that sufficient observation was given.

3-O-sulfated glucuronide derivative as a potential anti-dengue virus agent.

A series of 12 carbohydrate compounds were synthesized by introduction of a sulfated group at specific positions and evaluated for their activities against dengue virus (DENV) infection as well as binding to BHK-21 cells. 3-O-sulfated GlcA was active against DENV infection, whereas 2-O-sulfated GlcA and 3,6-di-O-sulfated Glc showed negligible activity. Persulfated compounds did not inhibit DENV infection. These results provided a rationale for designing sulfated carbohydrate compounds with low molecular mass as anti-DENV agents targeting E protein functions. 3-O-Sulfated GlcA showed no significant cytotoxicity at 1mM. The EC(50) value (120 µM) was lower than that of sucrose octasulfate (SOS), a small molecular weight inhibitor of DENV infection. Two negatively charged groups, 3-O-sulfate and 6-C-carboxylic acid, appear to be essential for anti-DENV activity. We performed docking study to investigate the binding potential of 3-O-sulfated GlcA with respect to DENV E protein. The docking study showed that distance and conformation of these negative charges on the carbohydrate may be suitable for association with three amino acid residues of E protein critically involved in virus adsorption (Lys295, Ser145, and Gly159). This interaction may competitively prevent functional DENV binding to receptor(s) on host cells. In conclusion, 3-O-sulfated GlcA is a chemical probe that may facilitate exploration of the molecular mechanisms underlying manifestations of dengue diseases.

Characterization of bisphenol A metabolites produced by Portulaca oleracea cv. by liquid chromatography coupled with tandem mass spectrometry.

The garden plant portulaca (Portulaca oleracea cv.) efficiently removes bisphenol A (BPA), an endocrine-disrupting chemical, from a hydroponic solution, but the molecular mechanisms underlying BPA metabolism by portulaca remain unclear. In this study, BPA metabolites converted by portulaca were analyzed by liquid chromatography coupled with tandem mass spectrometry. We observed the hydroxylation of BPA and the oxidization of it to quinone. Polyphenol oxidases are likely to contribute to BPA degradation by portulaca.

Alternative site for median nerve blockade allowing early functional rehabilitation after hand surgery.

PURPOSE: In this report we describe an alternative approach to catheter placement for continuous selective median nerve blockade. It spared the finger movements and therefore allowed early postoperative rehabilitation in a patient who underwent surgical repair of the index finger flexor tendon. CLINICAL FEATURES: A patient with a complicated history of traumatic index finger flexor tendon rupture, surgical repair, failed rehabilitation due to poor postoperative pain control, adhesion formation, and subsequent rerupture due to tenolysis was admitted for reconstructive surgery. This time, a continuous regional block was used. Although the insertion of a catheter at the wrist level would have spared the anterior interosseous branch of the median nerve and preserved finger movements, a more distant site had to be chosen to avoid proximity to the surgical wound. Therefore, under combined ultrasonography and neurostimulation guidance, the catheter was inserted in the proximal one-third of the patient's forearm distal to the branching-off point of the anterior interosseous nerve. Continuous ropivacaine infusion was initiated and maintained until being stopped on the afternoon of the third postoperative day, providing good analgesia without interfering with postoperative physiotherapy, which was successfully completed during this hospitalization. CONCLUSION: Placement of a catheter for continuous median nerve blockade in the proximal one-third of the forearm for effective postoperative pain-free rehabilitation after hand surgery should be considered in cases in which the surgical incision extends toward the patient's wrist. The block site can be readily identified by a combined use of ultrasonography and neurostimulation guidance.

[Case of sublingual hematoma following difficult laryngoscopy in a patient on anticoagulant therapy].

A 61-year-old man receiving anticoagulant therapy after the aortic valve replacement underwent emergency surgery for acute appendicitis. Although the patient was intubated with difficulty, no immediate complications were evident after extubation. On the evening of the first postoperative day he complained of a sore throat, difficulty in swallowing and dysphonia. The swelling of the tongue with right-side predominance and the swelling of the right neck were observed. The right-sided sublingual hematoma was confirmed by CT. The symptoms improved with steroid administration. The possibility of airway obstruction due to hematoma of the tongue should be considered in patients on anticoagulant therapy.

Antiviral activity of chondroitin sulphate E targeting dengue virus envelope protein.

Sulphated glycosaminoglycans such as heparin inhibit the early step of dengue virus infection through interaction with envelope (E) protein. Here, we found that chondroitin sulphate E (CSE), but not CSD, which contains the same degree of sulphation, inhibited dengue virus (DENV) infection of cells with adsorption. CSE significantly reduced infectivity of all dengue virus serotypes to BHK-21 and Vero cells. DENV preferentially bound to CSE immobilised on plastic plates. Also, virus binding to CSE or heparin was cross-inhibited by soluble CSE or heparin. These findings suggested that common carbohydrate determinants on CSE and heparin could be essential epitopes for interaction of DENV, and may be responsible for inhibition of the early steps of DENV infection. A recombinant E protein directly bound heparin and CSE, but not CSD, meaning that interaction of CSE with E protein contributes to the inhibitory action of this glycosaminoglycan. These observations indicate that a specific carbohydrate structure rather than polysulphation or addition of negative charges of the glycosaminoglycan molecule would be necessary for direct binding to DENV E protein. In conclusion, CSE showed antiviral activity as an entry inhibitor targeting E protein of dengue virus.

The common aspects of pathophysiolgy of alcoholism and depression.

Recent biological studies suggest the existence of the common pathophysiological aspects in alcoholism and depression. Postmortem studies have revealed the impairment of cAMP signaling in the patients with alcoholism. The similar alteration of cAMP signaling was also reported in postmortem brains of depressed patients. In this study, we supported the notion that neurogenesis would be essential in pathophysiology of both alcoholism and depression. Alcohol affected the function of neural stem cells (NSCs) and decreased neurogenesis at doses which did not affects cell survival, and treatment of antidepressant or moodstabilizer rescued the alcohol-induced suppression of neurogenesis. As the key mechanism of NSC differentiation change by ethanol and psychotropics, we focused on the transcriptional repressor, NRSF/REST activity change. Our in vitro studies demonstrated the NRSF/REST activation by ethanol and suppressive effect of antidepressants and lithium against its activation by ethanol. We further described the ERK reduction and ER stress in the cellular mechanism of NRSF/REST activation. All these findings suggested that cAMP-CREB cascade reduction and NRSF/REST activation may be common underlying mechanisms in the pathophysiology of alcoholism and depression.