Olivacine plus coadjuvants in the treatment of murine leukaemia

Administration of olivacine, 1,5-dimethyl-6H-pyrido[4,3-b] carbazole to mice inoculated with 10⁵, 10³ and 10² leukaemic cells (L₁₂₁₀) produced significant survival in all treated groups which was not altered by association with the naturally occurring nucleoside deoxycytidine. Treatment with olivacine plus heterologous serum caused leucocytosis due to neutrophilia, lymphocytosis and monocytosis, as opposed to treatment with olivacine alone, which produced leucopenia due to neutropenia and lymphocytopenia.     

Passive protection against Taenia saginata infection in cattle by a mouse monoclonal antibody reactive with the surface of the invasive oncosphere

The surface, excretory/secretory and intracellular compartments of Taenia saginata oncospheres were analysed by a combination of immunochemical techniques and by the use of selected hybridoma antibodies. The surface proteins of the oncospheres were directly iodinated by the lactoperoxidase technique and the intracellular and excretory/secretory components were labelled biosynthetically by culture in vitro with 35S-methionine. Analysis of radiolabelled proteins by SDS-PAGE revealed a restricted number of components in all of these three compartments. Mouse derived monoclonal antibodies directed against the oncospheral stage of this parasite demonstrated both stage specific and common determinants on the surfaces of the oncospheres and the metacestodes. One monoclonal (IgM) antibody, reactive with the oncosphere surface, which had a half life of 4.1 days when injected into calves, conferred protection against oral infection with T. saginata eggs. A monoclonal antibody reactive with a major secreted component did not confer passive protection.     

Expression of cellulose synthase-like (Csl) genes in insect cells reveals that CslA family members encode mannan synthases

Glucuronoarabinoxylan, xyloglucan, and galactomannan are noncellulosic polysaccharides found in plant cell walls. All consist of beta-linked glycan backbones substituted with sugar side chains. Although considerable progress has been made in characterizing the structure of these polysaccharides, little is known about the biosynthetic enzymes that produce them. Cellulose synthase-like (Csl) genes are hypothesized to encode Golgi-localized beta-glycan synthases that polymerize the backbones of noncellulosic polysaccharides. To investigate this hypothesis, we used heterologous expression in Drosophila Schneider 2 (S2) cells to systematically analyze the functions of the gene products of a group of Csl genes from Arabidopsis and rice (Oryza sativa L.), including members from five Csl gene families (CslA, CslC, CslD, CslE, and CslH). Our analyses indicate that several members of the CslA gene family encode beta-mannan synthases. Recombinant CslA proteins produce beta-linked mannan polymers when supplied GDP-mannose. The same proteins can produce beta-linked glucomannan heteropolymers when supplied both GDP-mannose and GDP-glucose. One CslA protein also produced beta-linked glucan polymers when supplied GDP-glucose alone. Heterologous expression studies of additional candidate glycan synthases in insect cells or other systems may help identify other noncellulosic polysaccharide biosynthetic enzymes.     

Location of <Emphasis Type="Italic">C. trachomatis</Emphasis> Inc Proteins during Expression of Their Genes in HeLa Cell Culture

Plasma constructions including genes encoding C. trachomatis inclusion membrane protein as composite proteins with reporter green fluorescent protein were obtained. After transfection of HeLa cell culture with the resultant plasmid constructions the location of inclusion membrane proteins in transfected cell was for the first time detected by confocal microscopy.__________Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 139, No. 5, pp. 562–566, May, 2005     

供体骨髓间充质干细胞干预非协调性异种肝移植免疫排斥反应的实验研究

目的研究骨髓间充质干细胞(MSCs)对非协调性异种肝移植排斥反应的免疫干预作用。方法密度梯度离心法分离培养豚鼠MSCs,流式细胞仪检测其膜表面CD34、CD45、CD44和CD29的表达,脂肪细胞诱导液培养诱导MSCs向脂肪细胞分化。豚鼠MSCs输注大鼠后检测不同时相免疫球蛋白IgG、IgM、IgA以及补体C3、C4的浓度变化。豚鼠和大鼠各30只随机分为3组,所有大鼠受体经环磷酰胺预处理,A组为MSCs输注组;B组为生理盐水输注组;C组为地塞米松输注组。各组行豚鼠大鼠原位肝脏移植手术,观察受体的存活情况及排斥反应情况。结果MSCs膜表面CD34阴性、CD45阴性、CD44阳性、CD29阳性。脂肪细胞诱导液培养后细胞内出现脂滴沉着。MSCs细胞悬液输注受体后各个时相的免疫球蛋白IgG、IgM和补体C3较输注前有显著降低(P<0.01),而IgA和补体C4与输注前相比无明显变化(P>0.05),输注后的免疫球蛋白IgG、IgM和补体C3各个时相点之间的浓度变化无显著差异(P>0.05)。肝脏移植模型A组受体存活时间为(431±27)min,较B组和C组的存活时间[(148±16)min、(141±22)min]显著延长(P<0.01)。A组受体超急性排斥反应发生较迟,程度轻。结论MSCs的鉴定可根据细胞形态,膜表面标志物和分化能力来确定,MSCs可干预异种肝脏移植的超急性排斥反应的发生。     

Xenotransplantation public perceptions: rather cells than organs

The aim of this study was to describe some of the factors that might play a role in influencing attitude to xenotransplantation: first, the consideration of receiving cells and tissue from xenotransplants in relation to whole xeno-organs; secondly, the fact that there is greater uncertainty regarding the result and risk of infection associated with xenotransplantation than with allotransplantation. We also describe the attitude to research on xenotransplantation, and the relationship between the attitude to receiving a xenotransplant and an allotransplant. Finally, we describe the attitude to xenotransplantation in relation to treatment for renal failure and waiting-time for allotransplantation. A questionnaire was sent to randomly selected members of the public aged 18 to 75 (n=1,000) and to all patients in the same age range who were waiting for kidney transplants in Sweden in the spring of 1998 (n=460). The response rate was 60% among the public and 87% among the patients. Both study groups were positive to a greater extent in their attitude to receiving cells and tissue than to receiving a whole organ such as a kidney. The response 'rather positive' to receiving organs was generally favored by the public, whereas the most generally favored response to receiving cells and tissue was 'very positive'. When there was suggested to be a greater uncertainty regarding the outcome with xenotransplantation compared with allotransplantation, the number of negative and uncertain respondents increased, both among the public and the patients. Eighty percent of the public and about 90% of the patients were in favor of continued research on xenotransplantation. Of those members of the public who responded, the attitude to receiving an organ from a human was positive in 86% of cases, with an emphasis on 'very positive'. There was a moderate relation between the attitude to receiving an organ from a human and to receiving a xenotransplant. Among the patients, there was no systematic or strong relation between the attitude to xenotransplantation and the kind of dialysis treatment they were on. Neither was there any systematic or strong relation to the waiting-time. The overall impression is that the attitude to xenotransplantation seems to be most influenced by whether the xenotransplant would involve whole organs or cells and uncertainty regarding the outcome.     

Activity and distribution of biogenic amines in structures of the thymus and spleen in response to injection of isologous and heterologous red cells

Injection of heterologous red cells was shown to lead after 15 min to the displacement of noradrenalin from efferent nerve endings, to a marked reorganization of the metabolism of the central follicular cells and the peripheral monoamine-containing cells of the marginal sinuses of the spleen, and cortical luminescent and mast cells of the thymus. Inversion of the medullary substance into the cortex was observed in the thymus. These changes increased in intensity until 2 h.Department of Histology and General Biology, Chuvash University. Laboratory for Transplantation of Organs and Tissues, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR V. V. Kovanov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 8, pp. 245–248, August, 1978.     

In vitro metabolism and drug interaction potential of a new highly potent anti-cytomegalovirus molecule, CMV423 (2-chloro 3-pyridine 3-yl 5,6,7,8-tetrahydroindolizine 1-carboxamide)

AIMS: To identify the enzymes involved in the metabolism of CMV423, a new anticytomegalovirus molecule, to evaluate its in vitro clearance and to investigate its potential involvement in drug/drug interactions that might occur in the clinic. METHODS: The enzymes involved in and the kinetics of CMV423 biotransformation were determined using pools of human liver subcellular fractions and heterologously expressed human cytochromes P450 (CYP) and FMO. The effect of CMV423 on CYP probe activities as well as on indinavir and AZT metabolism was determined, and 26 drugs were tested for their potential to inhibit or activate CMV423 metabolism. RESULTS: CMV423 was oxidized by CYP and not by FMO or cytosolic enzymes. The Km values for 8-hydroxylation to rac-RPR 127025, an active metabolite, and subsequent ketone formation by human liver microsomes were 44 +/- 13 microM and 47 +/- 11 microM, respectively, with corresponding Vmax/Km ratios of 14 and 4 microl min(-1) nmol(-1) P450. Inhibition with selective CYP inhibitors indicated that CYP1A2 was the main isoform involved, with some participation from CYP3A. Expressed human CYP1A1, 1A2, 2C9, 3A4 and 2C8 catalysed rac-RPR 127025 formation with Km values of < 10 microM, 50 +/- 21 microM, 55 +/- 19 microM, circa 282 +/- 61 microM and circa 1450 microM, respectively. CYP1B1, 2A6, 2B6, 2C19, 2D6, 2E1 or 3A5 did not catalyse the reaction to any detectable extent. CYP1A1 and 3A4 also catalysed ketone formation from rac-RPR 127025. In human liver microsomes, CMV423 at 1 and 10 microM inhibited CYP1A2 activity up to 31% and 63%, respectively, CYP3A4 activity up to 40% (10 microM) and CYP2C9 activity by 35% (1 and 10 microM). No effect was observed on CYP2A6, 2D6 and 2E1 activities. CMV423 had no effect on indinavir and AZT metabolism. Amongst 26 drugs tested, none inhibited CMV423 metabolism in vitro at therapeutic concentrations. CONCLUSIONS: CMV423 is mainly metabolized by CYP1A2 and 3A4. Its metabolism should not be saturable at the targeted therapeutic concentrations range (Cmax < 1 microM). CMV423 will probably affect CYP1A2 and 1A1 activities in vivo to some extent, but no other drug-drug interactions are expected.     

微囊化牛肾上腺嗜铬细胞脊髓蛛网膜下移植治疗慢性疼痛患者的初步观察

目的　观察海藻酸钠 -聚赖氨酸 -海藻酸钠 (APA)微囊化牛肾上腺嗜铬细胞 (BCC)异种移植对慢性顽固性疼痛患者的镇痛效应、作用持续时间及毒副作用。方法　用Sun氏微囊制作法将BCC包裹于APA微囊内 ,用常规腰穿法将 5ml微囊化BCC(5～ 7)× 10 6悬液注入患者L3～ 5蛛网膜下。结果　 2 0例中、重度慢性疼痛患者在 1或 2次注射后 ,疼痛迅速减轻 ,疼痛缓解率为 90 %。在未用任何免疫抑制剂的条件下 ,其中 3例停用镇痛药时间超过 2 0 0d。结论　APA微囊化BCC异种移植于慢性疼痛患者脊髓蛛网膜下 ,可安全、迅速、长时间、有效地发挥镇痛作用。     

One Step Enzyme Linked Immunosorbent Assay for Direct Estimation of Serum Cortisol

One step competitive enzyme linked immunosorbent assay (ELISA) for direct estimation of cortisol in human serum is described. Cortisol-3-O-carboxymethyl-oxime-bovine serum albumin (cortisol-3-O-CMO-BSA) was used as an immunogen and cortisol-21-hemisuccinate-horse radish peroxidase (cortisol-21-HS-HRP) was used as a tracer. to the cortisol antibody coated microtiter wells, standards or serum samples (25μ1) along with cortisol-HRP conjugate (100μ1) were incubated for 2 hours at 37°C. Bound enzyme activity was measured by, using TMB/H2O2 as a substrate. in this new strategy, chilled acetone stripped pooled human serum and sodium salicylate were used for preparing the standards and blocking the Cortisol binding globulin (CBG), respectively. the sensitivity of the assay was 0.28μg/100ml. the intraassay and interassay coefficient of variations (CVs) were ranged from 1.3% to 9.3% and 6.8% to 12.3 %, respectively. the analytical recoveries were 94% to 101.5%. the serum Cortisol values, obtained by this method were correlated well with those, obtained by radioimmunoassay; r=0.95 (n=52).