Hand-held monitor of sympathetic nervous system using salivary amylase activity and its validation by driver fatigue assessment

In order to realize a hand-held monitor of the sympathetic nervous system, we fabricated a completely automated analytical system for salivary amylase activity using a dry-chemistry system. This was made possible by the fabrication of a disposable test-strip equipped with built-in collecting and reagent papers and an automatic saliva transfer device. In order to cancel out the effects of variations in environmental temperature and pH of saliva, temperature- and pH-adjusted equations were experimentally determined, and each theoretical value was input into the memory of the hand-held monitor. Within a range of salivary amylase activity between 10 and 140 kU/l, the calibration curve for the hand-held monitor showed a coefficient with R(2)=0.97. Accordingly, it was demonstrated that the hand-held monitor enabled a user to automatically measure the salivary amylase activity with high accuracy with only 30 microl sample of saliva within a minute from collection to completion of the measurement. In order to make individual variations of salivary amylase activity negligible during driver fatigue assessment, a normalized equation was proposed. The normalized salivary amylase activity correlated with the mental and physical fatigue states. Thus, this study demonstrated that an excellent hand-held monitor with an algorithm for normalization of individuals' differences in salivary amylase activity, which could be easily and quickly used for evaluating the activity of the sympathetic nervous system at any time. Furthermore, it is suggested that the salivary amylase activity might be used as a better index for psychological research.     

糖化血红蛋白和糖化血清白蛋白联合检测在糖尿病中的筛查价值

目的：探讨糖化血红蛋白和糖化血清白蛋白联合检测在糖尿病中的筛查价值。方法：选取我院2012年3月至2013年7月842例进行糖尿病筛查的个体作为研究对象,通过检测研究对象HbA1c（糖化血红蛋白）、GA（糖化血清白蛋白）水平,分析HbA1c、GA和糖尿病的关系。结果：参加研究的受检者BMI为（24．32＋3．61）kg／m^2,收缩压为（128．11±18．25）mmHg,舒张压为（82．41±11．07）mmHg,空腹血糖（6．05±1．59）mmol／L,餐后2小时血糖（9．97±4．32）mmol／L,糖化血红蛋白（6．17±1.13）％,糖化血清白蛋白（17．82±4．53）％。NGT组、IGR组和DM组三组HbA1c、GA比较不全相等（P〈0．05）,且各组间两两比较也存在差异,差异均具有统计学意义（P〈0．05）;Pearson积差相关分析显示,研究对象HbA1c、GA、FPG（空腹血糖）、2hPG（餐后2小时血糖）这四个指标两两之间均呈正相关（P〈0．05）。结论：HbA1c和GA联合检测在糖尿病筛查中有应用价值,值得在实际工作中应用推广。     

Glycated Albumin at 4 Weeks Correlates with A1C Levels at 12 Weeks and Reflects Short-Term Glucose Fluctuations

Objective: Evaluate the performance of glycated albumin (GA) monitoring by comparing it to other measures of glycemic control during intensification of antidiabetic therapy.Methods: This 12-week, prospective, multicenter study compared the diagnostic clinical performance of GA to glycated hemoglobin A1C (A1C), fructosamine corrected for albumin (FRA), fasting plasma glucose (FPG), and mean blood glucose (MBG) estimated from self-monitoring of blood glucose (SMBG) and continuous glucose monitoring (CGM) in 30 patients with suboptimally controlled type 1 or 2 diabetes.Results: Mean A1C decreased from 9.5% to 8.1%. Mean SMBG correlated closely with CGM (Pearson r = 0.783 for daily estimates and r = 0.746 for weekly estimates, P<.0001). Both GA and FRA levels significantly correlated with changes from baseline in A1C and mean weekly SMBG (P<.001). The lowest observed median GA occurred at 4 weeks, followed by a small increase and then a slight reduction, mirroring changes in overall mean SMBG values. The median A1C fell throughout the treatment period, failing to reflect short-term changes in SMBG. A ≥1% reduction in GA at 4 weeks was significantly associated with a ≥0.5% change in A1C at 12 weeks (odds ratio &lsqb;OR] = 19.0, 95% confidence interval &lsqb;CI]: 1.4, 944, P = .018).Conclusion: In patients receiving glucose-lowering therapy, changes in GA at 4 weeks were concordant with changes in A1C at 12 weeks, and both GA and FRA more accurately reflected short-term blood glucose fluctuations than A1C.Abbreviations: A1C = glycated hemoglobin A1C ARIC = Atherosclerosis Risk in Communities CGM = continuous glucose monitoring FPG = fasting plasma glucose FRA = fructosamine corrected for albumin GA = glycated albumin MBG = mean blood glucose OR = odds ratio SMBG = self-monitoring of blood glucose     

Adjusting the number of spermatozoa in mini-straws by determination of the operative volume of bovine semen

The importance of the number of sperm per insemination on fertility has been well demonstrated in cattle. This number is usually calculated from the concentration in the extended semen and a theoretical value for the operative volume of semen delivered during insemination. The objective of this experiment was to investigate the usefulness of the measurement of the delivered volume of semen when estimating sperm numbers from frozen-thawed mini-straws, by comparing the results obtained with an analytical balance to the theoretical volume. The density of semen extended with Biociphos Plus and Triladyl was determined to be 1.033 g/ml using the gamma sphere method. This value was used to convert semen weight into operative volume. The effect of semen temperature at the time of weighing (37 degrees C versus 20 degrees C) was investigated on six semen batches, two technicians measuring the operative volume of 50 straws for each combination of temperature and semen batch (a total of 1200 weighings). The temperature effect was found to be insignificant, which allowed warm semen to be weighed before motility was assessed during routine quality control. The operative volume was then measured in straws routinely produced at 17 bovine Al centers (12-105 semen batches per center, mostly three straws from each batch, a total of 1912 measurements). The observed volumes were normally distributed around 198.7 microl, 98% measuring between 180 and 210 microl. The operative volume was significantly different among centers (from 192 to 205 microl, P < 0.0001) and among batches within centers (P < 0.0001). The S.D. among straws within batches was 3.4 microl. Some centers showed high variability in straw volume whereas others were more consistent. Determination of the operative volume of frozen-thawed mini-straws by weighing the delivered contents is an accurate method for estimation of the number of sperm per dose.     

Oxygen consumption and body temperature of active and resting honeybees

We measured the energy turnover (oxygen consumption) of honeybees (Apis mellifera carnica), which were free to move within Warburg vessels. Oxygen consumption of active bees varied widely depending on ambient temperature and level of activity, but did not differ between foragers (>18 d) and middle-aged hive bees (7-10 d). In highly active bees, which were in an endothermic state ready for flight, it decreased almost linearly, from a maximum of 131.4 microl O(2) min(-1) at 15 degrees C ambient temperature to 81.1 microl min(-1) at 25 degrees C, and reached a minimum of 29.9 microl min(-1) at 40 degrees C. In bees with low activity, it decreased from 89.3 microl O(2) min(-1) at 15 degrees C to 47.9 microl min(-1) at 25 degrees C and 14.7 microl min(-1) at 40 degrees C. Thermographic measurements of body temperature showed that with increasing activity, the bees invested more energy to regulate the thorax temperature at increasingly higher levels (38.8-41.2 degrees C in highly active bees) and were more accurate. Resting metabolism was determined in young bees of 1-7 h age, which are not yet capable of endothermic heat production with their flight muscles. Their energy turnover increased from 0.21 microl O(2) min(-1) at 10 degrees C to 0.38 microl min(-1) at 15 degrees C, 1.12 microl min(-1) at 25 degrees C, and 3.03 microl min(-1) at 40 degrees C. At 15, 25 and 40 degrees C, this was 343, 73 and 10 times below the values of the highly active bees, respectively. The Q(10) value of the resting bees, however, was not constant but varied in a U-shaped manner with ambient temperature. It decreased from 4.24 in the temperature range 11-21 degrees C to 1.35 in the range 21-31 degrees C, and increased again to 2.49 in the range 30-40 degrees C. We conclude that attempts to describe the temperature dependence of the resting metabolism of honeybees by Q(10) values can lead to considerable errors if the measurements are performed at only two temperatures. An acceptable approximation can be derived by calculation of an interpolated Q(10) according to the exponential function (V(O(2))=0.151 x 1.0784(T(a))) (interpolated Q(10)=2.12).     

Temperature-dependent fumigant activity of essential oils against twospotted spider mite (Acari: Tetranychidae)

Fumigant activity of 34 commercial essential oils was assessed on female adults and eggs of twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae) at three temperatures (5, 15, and 25 degrees C). Common thyme, cinnamon, and lemongrass oils were equally effective on twospotted spider mite adults showing 85.8-100% mortality at 5 and 10 microl/liter air at 25 degrees C. At a lower temperature of 15 degrees C, lemongrass and peppermint resulted in > or =90% mortality of adults at 10 microl/liter air. Only lemongrass was relatively active at 5 microl/liter air, at 15 degrees C. At 5 degrees C, lemongrass and peppermint caused significantly higher adult mortality than controls but only at 10 microl/liter air. Common thyme oil showed the highest ovicidal activity at 5 microl/liter air at 25 degrees C. Among the main components of common thyme and lemongrass oils, citral was lethal to twospotted spider mite adults at all tested temperatures. Carvacrol, thymol, and citral caused the same inhibitory effects on the hatch of twospotted spider mite eggs at 25 degrees C. However, citral was more active than other compounds to twospotted spider mite eggs at 15 degrees C. Therefore, we conclude that citral has the best potential for development as a fumigant against twospotted spider mite on agricultural products harvested late in the growing season.     

Expression of monocyte chemoattractant protein-1 by nonenzymatically glycated albumin (Amadori adducts) in vascular smooth muscle cells

The biological effects of Amadori adducts that are early nonenzymatically glycated protein on vascular cells were poorly defined. We examined the effect of glycated serum albumin (GA) on the expression of monocyte chemoattractant protein-1(MCP-1) that is an important chemokine recruiting monocyte to blood vessel. GA increased MCP-1 mRNA expression with a peak after 3 h of stimulation. The induction of MCP-1 by GA was dose-dependent. The MCP-1 mRNA expression by GA was completely inhibited by PD98059 and genistein that inhibit mitogen activated protein (MAP) kinase kinase and tyrosine kinase, respectively. N-Acetylcysteine, a potent antioxidant, also suppressed the GA-induced MCP-1 expression. These results suggest that GA induces production of reactive oxygen species and activates tyrosine kinase and MAP kinase in VSMC. Activation of these signals results in MCP-1 expression. GA-induced MCP-1 expression may be one of the mechanisms by which the diabetic patients suffer from accelerated atherosclerosis.     

Decreased Coomassie brilliant blue colour yield with glycated proteins.

Collagen, myosin and albumin were incubated for 7 days at 20 degrees C with fructose, ribose or glyceraldehyde. For thus-formed glycated proteins, quantities were determined by the Conway microdiffusion technique and by the colorimetric method based on Coomassie brilliant blue G-250 colour yield. It was found that when albumin was glycated with increasing amounts of glyceraldehyde, the colour yield was decreased by 7-33%. In collagen, myosin and albumin incubated with 0.5 mol/l fructose, 0.5 mol/l ribose or 0.1 mol/l glyceraldehyde, protein concentration was not changed, as proved by the Conway microdiffusion technique; the Coomassie brilliant blue G-250 colour yield was up to 50% lower, depending on the protein used, and was decreased much less when proteins were incubated with less sugar.     

Detection of micro-RNA by a combination of nucleic acid sequence-based amplification and a novel chemiluminescent pyrophosphate assay

Micro-RNA has attracted much attention as a biomarker for disease progression and malignancy. A compact, simple, rapid, and highly sensitive method is required to perform simple genetic analyses, such as point-of-care testing (POCT), at the clinic or bedside. Nucleic acid sequence-based amplification (NASBA) is a specific amplification method for a single-stranded RNA fragment that is useful for the highly sensitive detection of miRNAs. In this work, we developed a novel miRNA analytical system for POCT by combining the NASBA and chemiluminescence methods. Because the NASBA reaction is conducted at a constant temperature (41°C) and detection by chemiluminescence reaction does not require a light source, these methods could be combined to amplify 100 ng/assay miRNA. This combined miRNA detection method could be useful for the future development of compact POCT systems.     

Glycated albumin and diabetes mellitus

Background

Diabetes is a growing worldwide problem that is strongly associated with atherosclerosis. Screening and intervention for diabetes in the earliest stages are advocated for the prevention of diabetic complications and cardiovascular disease.

Scope of review

This review gives a background of and discusses the potential clinical utility of glycated albumin (GA) in diabetes.

Major conclusions

GA is a ketoamine formed via a non-enzymatic glycation reaction of serum albumin and it reflects mean glycemia over two to three weeks. GA can be used for patients with anemia or hemoglobinopathies for whom the clinically measured hemoglobin A1c level may be inaccurate. Because both serum and plasma samples can be used, GA can be analyzed from the same samples as common biological markers. GA is a useful marker for the screening of diabetes in a medical evaluation. It can be also used to determine the effectiveness of treatment before initiating or changing medications for diabetic patients. GA is potentially an atherogenic protein in the development of diabetic atherosclerosis.

General significance

GA measurement is useful as part of a routine examination to screen for both diabetes and atherosclerosis. This article is part of a Special Issue entitled Serum Albumin.     

糖化血清白蛋白检测对2型糖尿病患者短期血糖控制的疗效评价

目的：探讨糖化血清白蛋白（glycate dalbumin, GA）作为反映近期（2 周内）血糖控制总体水平的指标在糖尿病人群中的临床应用价值。方法：选取2010 年6 月-2013 年7 月间深圳市福田区中医院住院的2 型糖尿病（T2DM）患者323 例,测定空腹血糖（FPG）、餐后2 小时血糖（2hPG）、1 日指尖血糖谱均值（MBG）、GA、糖化血红蛋白（HbAlc）等,并对其中92 例住院近2 周的患者分别在入院后6 天及12 天复查上述指标。结果：GA 与HbAlc（r=0.791）,FPG（r=0.541）,2hPG（r=0.456）,MBG（r=0.401）,均呈正相关（P 均〈0.01）;患者入院6 天检测GA 为（27.1± 5.45）％,入院12 天为（22.77± 4.34）％,均显著低于入院第一天的（30.31±6.75）％（P〈0.01）;入院6 天和12 天,患者的GA 较入院时分别下降3.12％和7.54％（P〈0.01）;HbAlC 分别下降0.31％和0.78％,GA下降降幅显著大于HbAlc（P〈0.01）。结论：GA可及时、准确的反映短期（1～2 周）平均血糖的变化情况,并与长期血糖控制的金标准HbAlC 有良好的相关性,是监测糖尿病患者血糖控制的良好指标。     

Polymerase chain reaction of 2-kb cyanobacterial gene and human anti-alpha1-chymotrypsin gene from genomic DNA on the In-Check single-use microfabricated silicon chip

The microfabricated chip is a promising format for automating and miniaturizing the multiple steps of genotyping. We tested an innovative silicon biochip (In-Check Lab-on-Chip; STMicroelectronics, Agrate Brianza, Italy) designed for polymerase chain reaction (PCR) analysis of complex biological samples. The chip is mounted on a 1x3-in(2). plastic slide that provides the necessary mechanical, thermal, electrical, and fluidic connections. A temperature control system drives the chip to the desired temperatures, and a graphical user interface allows experimenters to define cycling conditions and monitor reactions in real time. During thermal cycling, we recorded a cooling rate of 3.2 degrees C/s and a heating rate of 11 degrees C/s. The temperature maintained at each thermal plateau was within 0.13 degrees C of the programmed temperature at three sensors. From 0.5 ng/microl genomic DNA, the In-Check device successfully amplified the 2060-bp cyanobacterial 16S rRNA gene and the 330-bp human anti-alpha(1)-chymotrypsin gene. The shortest PCR protocol that produced an amplicon by capillary electrophoresis comprised 30 cycles and was 22.5 min long. These thermal cycling characteristics suggest that the In-Check device will permit future development of a genotyping lab-on-a-chip device, yielding results in a short time from a limited amount of biological starting material.     

Fiber-optic biosensor to assess circulating phagocyte activity by chemiluminescence

We describe herein the construction of a novel computerized multi-sample temperature-controlled luminometer for a fiber array-based biosensor to monitor circulating phagocyte activity. It can perform simultaneously integral measurements of chemiluminescence emitted from up to six samples containing less than 0.5 microl whole blood while the samples and detector do not change their position during the measurement cycle. The optical fibers in this luminometer are used as both light guides and solid phase sample holders. The latter feature of the instrument design simplifies the assessment process of both the extra-cellular and the intra-cellular parts of the phagocyte-emitted chemiluminescence using the same system. We describe some examples or proof of principle for the use of the biosensor. This new technology may find use in a wide range of analytical luminescence applications in biology, biophysics, biochemistry, toxicology and clinical medicine.     

Cultural and clinical effectiveness of the 'QAAMS' point-of-care testing model for diabetes management in Australian aboriginal medical services

The national Quality Assurance for Aboriginal Medical Services (QAAMS) Program, in which point-of-care testing (POCT) for haemoglobin A(1c) (HbA(1c)) and urine albumin: creatinine ratio (ACR) is performed for diabetes management in 65 Australian Aboriginal medical services, is now embedded in the practice of diabetes care across Indigenous Australia. This paper documents the results of a detailed survey to assess levels of satisfaction with the QAAMS HbA(1c) Program among three key stakeholder groups-doctors, POCT operators and patients with diabetes. Both doctors and patients with diabetes agreed that the immediacy of POCT results contributed positively to patient care, improved the doctor-patient relationship, and made the patient more likely to be both compliant and self-motivated to improve their diabetes control. Both POCT operators and patients with diabetes reported improved satisfaction with their diabetes services after the introduction of POCT. The paper also provides evidence from two participating medical services that POCT has been an effective tool in improving the delivery of pathology services and clinical outcomes for both individuals and groups of patients with diabetes. A statistically significant reduction in HbA(1c) from 9.3% (+/- 2.0) to 8.6% (+/- 2.0) was observed in 74 diabetes patients 12 months after commencing POCT (p = 0.003, paired t-test). An improvement in the percentage of patients achieving glycaemic targets and a reduction in the percentage of patients with poor control was also observed in this group. These data provide evidence that the QAAMS POCT model delivers a culturally and clinically effective service for diabetes management in Aboriginal Australia.     

Glycated albumin induces superoxide generation in mesangial cells.

BACKGROUND/AIMS: Reactive oxygen species are involved in the pathogenesis of diabetic nephropathy. Amadori-modified glycated albumin modulates signaling pathways in mesangial cells that contribute to the development of diabetic nephropathy. However, the effects of glycated albumin on mesangial cell superoxide (O2-) production are unknown. Thus, we examined whether glycated albumin induces mesangial cell O2- generation and whether increased O2- production elicits cell growth. METHODS: Quiescent human mesangial cells (HMC) were exposed to bovine serum albumin (BSA) or glycated BSA (Gly-BSA) with or without diphenylene iodonium (DPI) or apocynin, inhibitors of NAD(P)H oxidase, GF109203X (GFX), a protein kinase C (PKC) inhibitor. RESULTS: Gly-BSA increased PKC activity, particularly PKC-alpha and -alpha1, within 15 min of incubation with HMC, which decreased to the control value at 2 h. Gly-BSA incubated with HMC increased O2- production by 2 times vis-á-vis BSA-treated cells. The Gly-BSA-induced increased O2- generation was suppressed by DPI or GFX. Gly-BSA significantly increased mesangial [3H]-leucine incorporation, whereas these processes were abrogated by DPI, apocynin or GFX. CONCLUSIONS: Gly-BSA induces PKC/NAD(P)H oxidase-dependent O2- production in HMC, which in turn results in cell hypertrophy. Thus, O2- induced by glycated albumin might cause mesangial cell alterations in diabetes participating in the pathophysiology of diabetic nephropathy.     

利用固定化黑曲霉单宁酶制备没食子酸的研究

用海藻酸钙载体包埋单宁酶,制备出转化五倍子单宁成没食子酸能力较好的固定化酶。研究了固定化条件和固定化单宁酶的部分性质,结果表明：最佳固定化条件为海藻酸钠90mg包埋单宁酶546u(3mL,182u/Ml),在1%～2%CaCl₂中作硬化处理;固定化单宁酶的最适温度为45℃,在10～50℃范围内稳定;其最适Ph值为6.5,在Ph5～7之间基本稳定;在此基础上,进行了没食子酸实验室克量级酶法制备实验,3次实验没食子酸产品的平均产率达到61%。和目前所用工业生产没食子酸的硫酸水解法相当,具有潜在的工业开发价值。     

Central CO-heme oxygenase pathway raises body temperature by a prostaglandin-independent way.

Recently, the carbon monoxide (CO)-heme oxygenase pathway has been shown to play an important role in fever generation by acting on the central nervous system, but the mechanisms involved have not been assessed. Thus the present study was designed to determine whether prostagandins participate in the rise in body temperature (T(b)) observed after induction of the CO-heme oxygenase pathway in the central nervous system. Intracerebroventricular (ICV) injection of heme-lysinate (152 nmol/4 microl), which is known to induce the CO-heme oxygenase pathway, caused an increase in T(b) [thermal index (TI) = 5.3 +/- 0.5 degrees C. h], which was attenuated by ICV administration of the heme oxygenase inhibitor ZnDPBG (200 nmol/4 microl; TI = 2.5 +/- 1.7 degrees C. h; P < 0.05). No change in T(b) was observed after intraperitoneal injection of the cyclooxygenase inhibitor indomethacin (5 mg/kg), whereas indomethacin at the same dose attenuated the fever induced by ICV administration of lipopolysaccharide (LPS) (10 ng/2 microl) (vehicle/LPS: TI = 4.5 +/- 0.5 degrees C. h; indomethacin/LPS: TI = 1.7 +/- 1.0 degrees C. h; P < 0.05). Interestingly, indomethacin did not affect the rise in T(b) induced by heme-lysinate (152 nmol/4 microl) ICV injection (vehicle/heme: TI = 4.5 +/- 1.4 degrees C. h; indomethacin/heme: TI = 4.2 +/- 1.0 degrees C. h). Finally, PGE(2) (200 ng/2 microl) injected ICV evoked a rise in T(b) that lasted 1.5 h. The heme oxygenase inhibitor ZnDPBG (200 nmol/4 microl) failed to alter PGE(2)-induced fever. Taken together, these results indicate that the central CO-heme oxygenase pathway increases T(b) independently of prostaglandins.     

Optical multichannel analyzer as a scanner for the ultracentrifuge. I. Single beam operation

The use of a commercial optical multichannel analyzer as a scanner for the ultracentrifuge is described. A uv-sensitive silicon vidicon tube serves as the light detector. The 5 by 12.5 mm surface of the vidicon is divided into 500 channels, scanned about 30 times/sec. An optical system was constructed which provided a reduced image of cell and counter-balance with the radius in the direction of channel number. The video signal from the vidicon is converted into digital data for each channel, the number of counts being proportional to the light intensity for that radial position. Storage registers are used to accumulate the counts for each channel, for any desired number of scans up to 9,999. For these studies, blue light from an H85C3 ac mercury lamp was used to illuminate the cell. Rotor and lamp pulses, visible on the real-time cathode-ray-tube monitor, were examined theoretically, and shown to have a negligible effect on the accumulated image. Only single-beam operation of the optical multichannel analyzer is described. The conversion of intensity data to absorbance as a function of radial position is described. The accuracy of the optical multichannel analyzer was verified by comparison of absorbances with values determined with a spectrophotometer.As a final test of the optical multichannel analyzer, the molecular weight of myoglobin was studied by sedimentation equilibrium. The reduced scatter in the experimental points permitted a more detailed analysis than usually performed. The final conclusion is that, even with single-beam operation at this early stage of development, the optical multichannel analyzer performs better than the double-beam, commercial scanner. Features already built into the optical multichannel analyzer lend themselves to automatic operation, either with a minicomputer or a specially constructed unit. Possible means of accomplishing double-beam and multiple cell operation are discussed.     

A novel method for measuring human lipoprotein lipase and hepatic lipase activities in postheparin plasma

The objective of this study was to establish a new lipoprotein lipase (LPL) and hepatic lipase (HL) activity assay method. Seventy normal volunteers were recruited. Lipase activities were assayed by measuring the increase in absorbance at 546 nm due to the quinoneine dye. Reaction mixture-1 (R-1) contained dioleoylglycerol solubilized with lauryldimethylaminobetaine, monoacylglycerol-specific lipase, glycerolkinase, glycerol-3-phosphate oxidase, peroxidase, ascorbic acid oxidase, and apolipoprotein C-II (apoC-II). R-2 contained Tris-HCl (pH 8.7) and 4-aminoantipyrine. Automated assay of lipase activities was performed with an automatic clinical analyzer. In the assay for HL + LPL activity, 160 microl R-1 was incubated at 37 degrees C with 2 microl of sample for 5 min, and 80 microl R-2 was added. HL activities were measured under the same conditions without apoC-II. HL and LPL activities were also measured by the conventional isotope method and for HL mass by ELISA. Lipase activity detected in a 1.6 M NaCl-eluted fraction from a heparin-Sepharose column was enhanced by adding purified apoC-II in a dose-dependent manner, whereas that eluted by 0.8 M NaCl was not. Postheparin plasma-LPL and HL activities measured in the present automated method had high correlations with those measured by conventional activity and mass methods. This automated assay method for LPL and HL activities is simple and reliable and can be applied to an automatic clinical analyzer.