The human serum paraoxonase—Polymorphism and specificity

Human serum contains EDTA‐sensitive (Ca⁺⁺‐dependent) and EDTA‐stable (albumin) paraoxonases which hydrolyse paraoxon, 0,0‐diethyl,0–4‐nitrophenyl phosphate.

In Caucasians the EDTA‐sensitive enzyme shows a genetically determined polymorphism which is governed by two alleles. In typical Mongoloid or Negro populations this polymorphism is expressed to a lesser degree, and in a few samples (e.g. Aborigines) it cannot be observed at all. The distribution of the activity of the EDTA‐stable (albumin) paraoxonase is unimodal.

Many authors supposed that paraoxonase is an arylesterase (EC 3.1.1.2) which means that it is also able to hydrolyse phenylacetate or β‐naphthylacetate. New investigations have shown that the human serum fractions splitting paraoxon can be separated from those hydrolysing phenylacetate and related substrates.

The polymorphism of the EDTA‐sensitive human serum paraoxonase can be applied to investigations concerning specificity. From this it becomes evident that this enzyme is rather specific. Already slight changes of the paraoxon molecule lead to a decrease of activity. On the other hand the enzyme seems to hydrolyse also phosphonic acid esters and 4‐nitrophenylacetate in the same way like paraoxon (polymorphism).

There is a linkage relationship of paraoxonase with cystic fibrosis and DNA markers. According to these results the (EDTA‐sensitive) paraoxonase locus is on the human chromosome 7.

A high serum paraoxonase level actually protects serum cholinesterase (EC 3.1.1.8) and has to be considered in biological monitoring of workers exposed to parathion.     

Patterns of serum calcium fractions,magnesium, phosphates and albumin in non‐insulin dependent diabetics (Nidd)

Blood level of calcium fractions (ionized and bound), phosphates, magnesium and albumin were measured in non‐insulin dependent diabetic patients (NIDD). Results were compared with healthy controls to elucidate the pathogenesis of probable changes of these elements. Total ionized calcium and magnesium showed significant decrease (p<0.001, p<0.05 and p<0.05) respectively. Phosphates showed non‐significant decrease while albumin reported nearly similar results. Positive correlation between inorganic phosphates and ionized calcium (r = 0.48, p<0.05) was observed only in healthy control group, not in diabetics. This may be attributed to alteration of ions excretions under diabetic condition through unknown mechanism. Additive increase of heparin sodium units in vitro to normal blood resulted in progressive fall in ionized calcium. Significant decrease was only observed upon addition of higher unit (5IU/ml). This must be taken into consideration biochemically for plasma determination of ionized calcium.     

全氟化合物同分异构体的环境行为及毒性效应研究进展

以全氟辛烷磺酸盐(PFOS)和全氟辛烷羧酸(PFOA)为代表的全氟化合物(PFASs)是一类新型持久性有机污染物,目前已经在全球自然环境、野生生物及人群中广泛检出,其环境与健康问题已引起人们的高度重视。传统的电氟化法生产工艺使PFASs产品存在碳链同分异构体。这些PFAS的同分异构体可能具有不同的环境行为和毒理效应。随着分析方法的逐渐成熟,目前,国外的学者已经在该研究领域进行了一些探索性的研究,并取得了一定的进展,而我国在该领域的研究相对较少。以PFOS和PFOA为例,在介绍了PFAS同分异构体的来源、命名和世界各地生产厂家的异构体组成等基本信息的基础上,还系统介绍了环境介质中PFAS同分异构体的分析方法、环境迁移转化差异及源分析研究;生物和人体的生物累积性及毒理学差异等;并对目前存在的问题进行了讨论,为今后PFAS同分异构体的环境问题研究提供相应的参考。     

增塑剂DIDP对肝脏氧化应激损伤及VitE拮抗作用研究

研究增塑剂邻苯二甲酸二异癸酯(didecyl phthalate, DIDP)致雄性小鼠肝损伤作用及其机理。以雄性BALB/c小鼠为受试动物,随机分为7组,包括溶剂对照组(生理盐水)、4个DIDP染毒组(0.15、1.5、15和150 mg·kg~(-1))、维生素E(vitamin E, VitE)(100 mg·kg~(-1))处理组和DIDP+维生素E处理组(150 mg·kg~(-1)DIDP+100 mg·kg~(-1)VitE),连续灌胃14 d。以肝组织匀浆测定活性氧(reactive oxygen species, ROS)、还原型谷胱甘肽(glutathione, GSH)、丙二醛(malondialdehyde, MDA)和细胞凋亡因子半胱氨酸天冬氨酸蛋白酶3(cysteine aspartic proteinase 3, Caspase-3)水平。采用动物自动生化分析仪检测肝功能指标血清中丙氨酸氨基转移酶(alanine aminotransferase, ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase, AST)、白蛋白(albumin, ALB)水平,并同时观察肝组织的病理变化与荧光染色结果。随着DIDP染毒剂量的增加,小鼠肝组织ROS、MDA和Caspase-3含量逐渐上升,血清ALT和AST水平也逐渐上升,GSH含量逐渐降低,血清ALB水平也逐渐降低,差异具有统计学意义(P 0.05,P 0.01); VitE处理组ROS、MDA和Caspase-3含量相应降低,血清ALT和AST水平也相应降低,GSH含量逐渐上升,血清ALB水平也相应上升。小鼠肝组织形态观察结果表明,随着DIDP染毒剂量的增加,小鼠肝组织的病理损伤程度呈上升趋势。研究表明,较高剂量(≥15 mg·kg~(-1))的DIDP能造成小鼠的肝脏损伤与细胞凋亡,抗氧化剂VitE可使肝脏损伤与细胞凋亡减轻,对小鼠肝组织起保护作用,说明氧化应激介导了DIDP对机体的损伤。     

Development of in-house ELISA for detection of chloramphenicol in bovine milk with subsequent confirmatory analysis by LC-MS/MS

This study was undertaken to develop and validate direct competitive ELISA for the determination of chloramphenicol residues in bovine milk. Antisera and an enzyme-tracer for chloramphenicol were prepared and used to develop an ELISA with inhibition concentrations, IC₂₀ and IC₅₀, of 0.09 and 0.44 ng mL^?1, respectively. Milk samples were spiked with standards equivalent to 0, 0.2, 0.3, 0.5, 1.0 &; 1.5 ng mL^?1 and extracted in methanol. The mean recoveries were found to be 73–100% with coefficient of variance 7–11%. The decision limit (CCα) and detection capability (CCβ) were calculated as 0.10 and 0.12 ng mL^?1, respectively. The results were found comparable with the commercial ELISA, having recoveries of 87 to 100%, CCα 0.09 ng mL^?1 and CCβ 0.12 ng mL^?1. As per Commission Decision 2002/657/EC, in-house ELISA was further validated by using LC-MS/MS. Mass spectral acquisition was done by using electrospray ionization in the negative ion mode applying single reaction monitoring of the diagnostic transition reaction for CAP (m/z 152, 194 and 257). The calibration curve showed good linearity in concentrations from 0.025 to 1.6 ng mL^?1 with correction coefficient 0.9902. The mean recoveries were found to be 88 to 100%. The CCα was calculated as 0.057 ng mL^?1 and CCβ 0.10 ng mL^?1. Since CCα and CCβ are less than half of the MRPL (0.15 ng mL^?1), the test was found suitable for screening and quantification of CAP residues in bovine milk samples. Results of surveillance studies indicated that out of 31 analyzed milk samples, 12.9% samples were found with CAP residues but only 3.2% samples were declared positive with maximum concentration 0.31 ng mL^?1, slightly above the MRPL.     

干旱因素对尿、血清和唾液含氟量的影响

我们曾就干旱因素影响两个地方性氟中毒(地氟病)病区人群的某些指标作过报道。本文重点讨论其影响同一人群尿、血清和唾液含氟量情况,旨在为地氟病防治研究提供参考。     

Determination of maternal serum acetylcholinesterase in pregnancies with fetal neural tube defects

We have investigated the occurrence of acetylcholinesterase (AChE) (E.C.3.1.1.7) in fetal serum, amniotic fluid and maternal serum using an immuno-chemical assay-technique employing both polyclonal and monoclonal antibodies. Fetal serum had increased amounts of AChE, which is due to an increase in the 10.5S form of the enzyme. This form was also found in amniotic fluids of pregnancies with a fetal neural tube defect (NTD), but not in normal amniotic fluid. The increase in amniotic fluid AChE was however, not reflected in the maternal serum.     

Raised maternal serum alpha-fetoprotein in the absence of fetal abnormality-placental findings. A quantitative morphometric study

Ten placentae from pregnancies proceeding to term from mothers who on routine screening at 16–18 weeks gestation were found to have a raised serum AFP but no increase in amniotic fluid AFP and no fetal abnormality, were studied using morphometric techniques. The results were compared with 20 placentae from normal term pregnancies where the maternal serum AFP level was not elevated. The mean total placental volume, volume of parenchyma and villous surface area were increased in the placentae associated with a raised maternal serum AFP. More of these placentae were infarcted and the fetal-placental weight ratio was significantly lower. The hypothesis that elevation of maternal serum AFP level is related to the increase in placental size is addressed.     

Four-marker serum screening for Down's syndrome

The value of measuring the separate sub-units of human chorionic gonadotrophin (free α-hCG and free β-hCG) instead of total hCG together with alpha-fetoprotein (AFP) and unconjugated oestriol (uE₃) was examined to determine the effect on the performance of serum screening for Down's syndrome between 15 and 22 weeks of pregnancy. The study was based on stored serum samples relating to 75 singleton pregnancies with fetal Down's syndrome and 367 unaffected singleton pregnancies, matched for maternal age, gestational age, and duration of storage of the serum sample, supplemented by data from 970 white women with unaffected pregnancies. Using the four serum markers AFP, uE₃, free β-hCG, and free α-hCG, in addition to maternal age, 65 per cent of Down's syndrome pregnancies were detected for a 5 per cent false-positive rate compared with 59 per cent with the conventional triple test (AFP, uE₃, total hCG with maternal age). If gestation was based on an ultrasound scan examination, the detection rate was 72 per cent using the four serum markers compared with 67 per cent with the triple test. As an alternative illustration, if the detection rate was kept at 60 per cent and gestation was estimated by an ultrasound scan examination the four-marker test reduced the false-positive rate by one-third from 3 per cent using the triple test to 2 per cent with the four-marker test. Screening performance was hardly affected by adjusting marker levels for maternal weight. The four-marker test is, both from a medical and from a financial perspective, the most effective method of prenatal screening for Down's syndrome suitable for routine use.