Computational chemistry in biotransformational and toxicological research

Conclusion In conclusion, our studies have shown that computational chemistry techniques are very useful to elucidate mechanisms for the oxidation of substrates by cytochromes P-450 and to gain insight into the active site structures of these enzymes. In a similar way, these techniques may be employed in other fields concerning biotransformational and toxicological research and can thereby reduce the use of experimental animals. When compared toin vivo andin vitro approaches, thein computro approach might be of great use in biotransformational and toxicological research.     

Monoamine oxidase in rat reticulocytes: Subcellular localization and identification of isoenzymes

Summary After chemically induced reticulocytosis in rats by treatment with acetyl-phenylhydrazide, monoamine oxidase (MAO) activities were determined in erythrocyte preparations of these animals. Studies on subcellular fractions obtained by differential centrifugation showed that the enzyme activity of rat reticulocytes is a classical mitochondrial MAO.The patterns of inhibition produced by clorgyline (A-type MAO), deprenil (B-type MAO) and pargyline or tranylcypromine (both types of MAO) in reticulocytes were determined in vitro using tryptamine as a substrate for both types of MAO and phenylethylamine as a substrate for the B-type. The results indicate that both A-type ( 75%) and B-type ( 25%) MAO are present in rat reticulocytes; while tryptamine was mainly deaminated by the A-type enzyme, both types of MAO were shown to contribute to the deamination of phenylethylamine. These findings were confirmed in investigations on the thermostabilities of the tryptamine and phenylethylamine deaminating activities of rat reticulocyte MAO.This work was supported by a grant from the Deutsche ForschungsgemeinschaftPreliminary accounts on this work were presented at the 16. Frühjahrstagung der Deutschen Pharmakologischen Gesellschaft (Quiring and Hubertus, 1975) and at the 6th International Congress of Pharmacology (Quiring et al., 1975 a)The experimental work presented in this paper includes the work done by S. H. for his doctoral thesis     

Über eine Beziehung zwischen sog. Dünndarmphosphatase und Pseudocholinesterase im Serum bei Lebercirrhose

Zusammenfassung An Hand von 30 Patienten mit histologisch gesicherter aktiver Lebercirrhose wird über eine statistisch signifikante Korrelation (=–0,395;x ²=4,692; 1 FG;p<0.05) zwischen erniedrigter Cholinesterase-Aktivität und Vermehrung der sog. Dünndarmphosphatase im Serum berichtet.

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Correlation between a decrease of pseudocholinesterase and an increase of the small intestine phosphatase in the serum of patients with active liver cirrhosis

Summary Sera of 30 Patients with histologically active liver cirrhosis have been investigated.A statistically significant correlation (=–0,395;x ²=4.692; 1 GF;p<0.05) was found between a decrease of pseudocholinesterase and an increase of the so called small intestine phosphatase in the sera of patients with active liver cirrhosis.

     

Serum adenosine deaminase and procalcitonin concentrations in neutropenic febrile children with acute lymphoblastic leukaemia

Abstract Neutropenia as a state of immunosuppression is probably the major problem in patients suffering from acute lymphoblastic leukaemia undergoing intensive chemotherapy. Fever is frequent in neutropenic patients and often related to infection. Clinically, the presence of infection in patients with neutropenia may be difficult to establish, because there are usually few signs of infection. The aim of this work was to study sensitive markers for early diagnosis of microbial infection in neutropenic children undergoing intensive chemotherapy as a treatment for acute lymphoblastic leukaemia. The study included three groups (A, B and C) of children with acute lymphoblastic leukaemia and neutropenia. Group A consisted of 29 children with febrile neutropenia and microbial infection, aged 1–14 years (5.8±2.9), 11 boys and 18 girls; Group B of 38 children with febrile neutropenia without microbial infection, aged 2–14 years (6.8±3.1), 14 boys and 24 girls; and Group C of 53 children with neutropenia without fever and without infection, aged 1–14 years (5.9±2.1), 21 boys and 32 girls. Blood samples were collected upon admission and before the start of any antimicrobial treatment. The samples were used for blood culture, serological tests, leukocyte count and analysis of levels of C–reactive protein, procalcitonin, total adenosine deaminase (ADA) activity and its isoenzymes, ADA–1 and ADA–2. According to our results the procalcitonin levels and total ADA activity discriminated best between neutropenic febrile (Groups A and B) and neutropenic afebrile episodes (Group C). In conclusion, this study suggests procalcitonin and total ADA activity as two easily measurable and cost effective markers for the assessment of immune response in febrile neutropenic patients with acute lymphoblastic leukaemia.     

血清心肌肌钙蛋白Ⅰ测定在心肌梗死中的应用

目的 为探讨血清心肌肌钙蛋白Ⅰ测定在心肌梗死中的诊断价值。方法 采用磁微粒子化学发光技术定量检测急性心肌梗死（ＡＭＩ）患者、下沉脸血清心肌肌钙蛋白１（ＣＴｎ１）,并与肌酸激酶同工酶（ＣＫ－ＭＢ）活性比较。结果 ｃＴｎ１对ＡＭＩ患者敏感性９８．４％,阳性率在发病后４小时内达４４．４％,７天阳必有达３８．４％。结论 其阳性出现时间早持续时间长,ｃＴｎ１灵敏度及特异性明显优于ＣＫ－ＭＢ活性,是高度灵敏、     

Adenosine Deaminase Activity and its Isoenzyme Pattern in Patients with Juvenile Rheumatoid Arthritis and Systemic Lupus Erythematosus

Adenosine deaminase (ADA) is involved in purine metabolism and plays a significant role in the mechanisms of the immune system. The aim of this study was to investigate the activity of total ADA (tADA) and its isoenzymes ADA1 and ADA2 in serum and peripheral blood lymphocytes (PBLs) of children with juvenile rheumatoid arthritis (JRA) and systemic lupus erythematosus (SLE) in different phases of the diseases. The study comprised 34 patients with rheumatic disease, 24 with JRA and 10 with SLE, and 64 healthy controls. The tADA activity and its isoenzymes were measured in serum and PBLs of all patients by the method of Giusti and by the presence or absence of EHNA (erythro-9-(2-hydroxy-3-nonyl)adenine) during the active phase of the disease (before treatment), as well as during remission and relapse. Our data show that increased tADA activity in the serum and PBLs of patients with JRA and SLE is correlated mainly to increased levels of ADA2 activity in serum and ADA1 activity in PBLs. It also closely correlates with clinical disease activity and relapse. The cause of this increased tADA/ADA2 activity in serum and tADA/ADA1 activity in PBLs in JRA and SLE remains to be elucidated. Nevertheless, it may be noted that the measurement of tADA activity, together with ADA2 activity in serum and tADA with ADA1 activity in PBLs, could offer a biochemical approach to the assessment of the pathophysiology of JRA and SLE. Also, tADA and its isoenzymes could be used as alternative parameters representing disease activity. Received: 31 July 2000 / Accepted: 5 June 2001     

Morphological and genetic characterization of Echinococcus granulosus in the Slovak Republic

The present study was undertaken to characterize more thoroughly aetiological agent of cystic echinococcosis that still represents a health problem in the Slovak Republic. Sequencing of the mitochondrial ND1 gene, Random Amplified Polymorphic DNA (RAPD) analysis and isoenzyme analyses were used to characterize genetically protoscolices of 37 isolates from pigs and cattle and two isolates from humans collected in different regions of the Slovak Republic. Solely the G7 genotype (pig strain) was identified by ND1 sequences in all 14 pigs isolate examined by this method. This genotype was also found in two human patients and in two cattle. The exclusive finding of the G7 variant has implications for the implementation of control strategies given its unique developmental and biological properties. Despite striking morphological variability, a low degree of the intra-strain variability was found in the Slovak Echinococcus granulosus isolates with the genetic methods used. This was particularly associated with the shape and size of hooks, number of testes and shape of ovaries and uterus. GPI and MDH enzymes were found suitable for the discrimination of G7 and G1 (=cosmopolitan sheep strain) variant.     

肿胀激活状态下蛋白激酶C同工酶亚型在胃癌耐药细胞系的亚细胞分布变化及其意义

目的 探讨肿胀激活状态下，传统型蛋白激酶C（cPKC)同工酶亚型在胃癌SGC7901及其耐药细胞系SGC7901/VCR的表达，亚细胞分布变化及其意义。方法 采用免疫荧光及免疫印迹方法，观察正常状态及持续低渗灌流不同时间cPKC同工酶亚型的亚细胞分布变化，利用免疫荧光双标记及共聚焦显微镜技术观察PKCα与-P-糖蛋白（Pgp)的共表达。结果 在正常状态下，cPKC的4种同工酶亚型在胃癌SGC7901及其耐药细胞SCG7901/VCR细胞质及细胞膜均有表达：PKCα在耐药细胞表达呈强阳恬药敏细胞表达呈强阳性，PKCγ在耐药细胞及药敏细胞表达均呈强阳性。持续低渗灌流，在耐药系10min已发现有PKCα、PKCγ的移位及细胞体积的增大；30minPKCα几乎全部移位至细胞膜上，PKCγ部分移位至细胞核内，细胞体积较前增大1-2倍；60min时PKCα及PKCγ基本恢复至正常位置。细胞体积也恢复正常；在药敏细胞系10-20minPKCα及PKCγ几乎全部一笔勾销 位至细胞膜上，PKCγ部分移位至细胞核内，细胞体积也恢复正常；在药敏细胞系10-20minPKCα几乎全部移位至细胞膜上，PKCγ部分移位至细胞核内，细胞体积较前增大1-2倍；40minPKCα及PKCγ基本恢复至正常位置，细胞体积也恢复正常，而PKCβ1及PKCβα在耐药细胞及药敏细胞的亚细胞分布无明显变化。共聚焦显微镜提示PKCα及PKCγ同工酶亚型参与了Pgp与低渗刺激下细胞共表达，以耐药细胞表达明显。结论 只有PKCα及Pgp表达量有关，而PKCβ1及PKCβⅡ可能与这一过程无关。βββ     

Isoenzyme analysis ofEntamoeba histolytica for evaluation of pathogenicity: Detection of zymodeme XIX in South America

Summary Authors report the results of the isoenzyme analysis of strains ofEntamoeba histolytica isolated from international travellers. This recently developed method allows the detection of pathogenic strains by evaluating the electrophoretic mobility of cytoplasmic enzymes and was proved to be more reliable and quickly feasible than previous ones. The experience reported refers to three strains isolated from travellers coming from Ecuador, Brazil and Indonesia, respectively; the zymodeme XIX (in accordance with the Sargeaunt’s classification) was identified in all the three cases. This zymodeme has been first detected in 1981 and should currently be considered rare; moreover, it has never been previously reported from the Americas.