Depolymerized holothurian glycosaminoglycan (DHG), a novel alternative anticoagulant for hemodialysis,is safe and effective in a dog renal failure model

BACKGROUND: Depolymerized holothurian glycosaminoglycan (DHG) is a new agent with anticoagulant properties quite different from those of unfractionated heparin (UFH) and low-molecular-weight heparin (LMWH) in terms of antithrombin III-dependency, and exerts an antithrombotic effect with less bleeding than UFH and LMWH in vivo. In this study, the anticoagulant and hemorrhagic effects of DHG were investigated on hemodialysis in a dog model of renal failure and compared with those of UFH, LMWH, and nafamostat mesilate (FUT). METHODS: The dog renal failure model was prepared by 7/8 renal artery ligation. Effectiveness was based on completion of 3-hour hemodialysis, no marked clot deposition in the extracorporeal circuit, and permeability of blood urea nitrogen (BUN) and creatinine. Template bleeding was measured by determining the hemoglobin content of the blood from the wound. RESULTS: DHG induced no major bleeding or clot formation during 3-hour hemodialysis, in contrast to UFH and LMWH, each of which induced marked bleeding. These glycosaminoglycans (GAGs) were equally effective in decreasing plasma levels of BUN and creatinine. On the other hand, dogs treated with FUT failed to complete 3-hour hemodialysis. These anticoagulants prolonged activated partial thromboplastin time (APTT) to different extents and GAGs prolonged thrombin clotting time markedly but FUT did not. CONCLUSION: Our findings suggest that thrombin clotting time prolongation can contribute to prevention of clot formation in extracorporeal circuits, and the non-antithrombin III-dependent activities of DHG may be related to its low risk of hemorrhage for hemodialysis. DHG appears to be promising as an alternative anticoagulant with low risk of hemorrhage for hemodialysis.     

Successful excision of a right ventricular fibroma associated with ventricular tachycardia. Report of a six year survival

A right ventricular (RV) tumor manifested by ventricular tachycardia (VT) accompanied by syncopal attacks was found in a 14 year old boy by two-dimensional echocardiography. Surgery was performed on February 10, 1981, with the aid of a cardiopulmonary bypass. The tumor was completely removed as a mass from the anterolateral portion of the RV wall. The wall was closed directly without any patch. The mass was 60 gm in weight and 7 by 4 by 4 cm in size. Fibroma was diagnosed by pathological study. The patient is doing well 6 years postoperatively. We conclude that two-dimensional echocardiography should be used to exclude cardiac tumors such as fibroma in young patients who have VT.     

贵州老鹰茶提取物体外抗单纯疱疹病毒1型活性的实验研究

目的研究贵州老鹰茶甲醇提取物体外抗单纯疱疹病毒1型(HSV-1)活性。方法采用细胞病变效应(Cytopatho-genic effect,CPE)法与空斑减数实验(Plaque reduction assay,PRA)测定甲醇提取物抗HSV-1活性,计算其空斑抑制率和半数抑制浓度(50%inhibiting concentration,IC50),并从药物对病毒的直接灭活作用、对病毒吸附的影响及对病毒穿膜的影响3个方面初步探讨其抗HSV-1的活性机理。结果贵州老鹰茶甲醇提取物能明显抑制HSV-1的致病变作用,其IC50为12.02μg/ml。药物主要是影响病毒对细胞的吸附,对病毒也有一定的直接灭活作用。结论贵州老鹰茶甲醇提取物有显著的抗HSV-1活性,初步推测其抗HSV-1活性的机理是作用在HSV-1和受体结合,侵入Vero细胞阶段。     

Potential for the replication of the betanodavirus redspotted grouper nervous necrosis virus in human cell lines

Summary The determination of the host ranges of viruses is important because of the possible emergence of infectious agents, which may result from the zoonotic transmission of animal viruses to humans. The family Nodaviridae, whose members are non-enveloped, positive-stranded bipartite RNA viruses, is comprised of the genera Alphanodavirus and Betanodavirus, whose members predominantly infect insects and fish, respectively. The alphanodaviruses can also infect suckling mice and suckling hamsters, resulting in paralysis and death. Pigs near the site of isolation of the Nodamura virus (NoV), an alphanodavirus, have been reported to have high levels of NoV neutralizing antibody, suggesting that they may be part of the natural host range of this virus. Betanodaviruses are the causative agents of viral nervous necrosis, which occurs in several species of fish. However, little is known regarding the mechanism of infection of these viruses. Whether betanodaviruses can infect hosts other than fish remains unclear. In this study, we examined the possibility that a betanodavirus, redspotted grouper nervous necrosis virus (RGNNV), can infect human cell lines and showed that this virus can attach to the cells but cannot penetrate them, although human cells can support the replication of the betanodavirus when viral RNAs are transfected. The betanodavirus in its present form cannot infect human cells. Correspondence: Nobuyuki Kobayashi, Laboratory of Molecular Biology of Infectious Agents, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan     

Depolymerized holothurian glycosaminoglycan with novel anticoagulant actions: antithrombin III- and heparin cofactor II-independent inhibition of factor X activation by factor IXa-factor VIIIa complex and heparin cofactor II-dependent inhibition of thrombin

The inhibition mechanism of a polysaccharide anticoagulant, depolymerized holothurian glycosaminoglycan (DHG), was examined by analyzing its effects on the clotting time of human plasma depleted of antithrombin III (ATIII), of heparin cofactor II (HCII), or of both heparin cofactors. The effect exerted by this agent on the activation of prothrombin and factor X in purified human components were also examined and all effects were compared with those of other glycosaminoglycans (GAGs). The capacity of DHG to prolong activated partial thromboplastin time was not reduced in ATIII-depleted, HCII- depleted, HCII-depleted, or ATIII- and HCII-depleted plasma, whereas its capacity to prolong prothrombin time and thrombin clotting time was reduced in HCII-depleted plasma. DHG inhibited the amidolytic activity of thrombin in the presence of HCII with a second order rate constant of 1.2 x 10(8) (mol/L)-1 min-1. These results indicated that DHG has two different inhibitory activities, one being an HCII-dependent thrombin inhibition and the other an ATIII- and HCII-independent inhibition of the coagulation cascade. The heparin cofactors- independent inhibitory activity of DHG was investigated in the activation of prothrombin by factor Xa and in the activation of factor X by tissue factor-factor VIIa complex or by factor IXa. DHG significantly inhibited the activation of factor X by factor IXa in the presence of factor VIIIa, but not in the absence of factor VIIIa. The interaction between DHG and factors IXa, VIIIa, and X was investigated with a DHG-cellulofine column, on which DHG had strong affinity for factors IXa and VIIIa. These findings show that the heparin cofactors- independent inhibition exhibited by DHG was caused by inhibition of the interaction of factor X with the intrinsic factor Xase complex, probably by binding to the factor IXa-factor VIIIa complex.     

In vitro activities of piperacillin against beta-lactamase-negative ampicillin-resistant Haemophilus influenzae

The in vitro activities of piperacillin (PIP) against beta-lactamase-negative ampicillin (AMP)-resistant (BLNAR) Haemophilus influenzae were compared with those of cefotaxime (CTX) and ceftriaxone (CRO), and the potency of PIP as therapy for meningitis caused by BLNAR is also discussed. PIP showed good activity (MIC at which 90% of strains are inhibited, 0.25 micro g/ml) against 69 BLNAR strains, and its activity was comparable to that of CRO and superior to that of CTX. No significant correlation was observed between the MICs of PIP and CTX or CRO or AMP, whereas a high correlation was observed between the MICs of CTX and CRO. In the killing study, PIP showed potent bactericidal activity compared with those of CTX and CRO. By microscopic examination, PIP caused the formation of a spindle and short filamentous cells with bulges and induced cell lysis in BLNAR strains, while treatment with CTX and CRO resulted in the formation of large, spherical cells without any obvious lysis. The affinity of Bocillin FL, a fluorescent penicillin used for determination of the 50% inhibitory concentration (IC(50)s) for penicillin-binding proteins (PBPs), to PBPs 3a and 3b of BLNAR strains was drastically decreased compared with that to an AMP-susceptible strain (ATCC 33391). In the case of the BLNAR strains, the IC(50)s for PBPs 1a, 1b, and 2 were similar to those for the PBPs of ATCC 33391. Since the affinity of binding to PBPs 3a and 3b of the BLNAR strains decreased drastically, the second targets among the PBPs were PBP 2 for PIP, PBP1 (1a and 1b) for CTX and CRO. In conclusion, PIP showed excellent activities against BLNAR strains in a manner different from those of cephem antibiotics, suggesting that it could be a candidate therapeutic agent for the treatment of meningitis caused by BLNAR strains.