Serum collection

The criterion for purity was a single band on a heavily loaded SDS-polyacrylamide electrophoresis gel. Antibodies were raised in rabbits in response to three intravenous injections each of 2mg. of laccase A with Freunds adjuvant, at 2 weekly intervals. Blood was removed 7d after the last injection and serum collected after coagulation of the blood cells. 

This type of rotation error has also been observed when embryos are cultured with serum collected from ketamine-anesthetized rats (personal observation). Therefore, it is recommended to evaluate this effect against the others in the treatment group, as it could have non-treatment-related origins. 

Free fatty acids are elevated in the plasma of obese patients and are known to cause muscle and liver insulin resistance. The Wako HR series NEFA-HR(2) is an in vitro enzymatic colorimetric method assay for the quantitative determination of non-esterified fatty acids (NEFA) in serum. Perform the assay on serum collected from mice fasted for a period greater than 4 h, but less than 16 h. Perform the test on samples immediately after collection, without freezing. Also note that hemolysis in the serum samples may interfere with the assay. 

The first group underwent a conventional immunization procedure (6) in which one half mg of ECPs were administered subcutaneously in Complete Freunds Adjuvant (CFA) on day one and in Incomplete Freunds Adjuvant (ICFA) on day 7. The rabbits were boosted every 14 days for 160 days and serum collected 7 days after each boost. 

H. Tsuchihashi, M. Katagi, M. Nishikawa and M. Tatsuno, Identification of metabolites of nerve agent VX in serum collected from a victim, J. Anal. Toxicol., 22, 383-388 (1998). 

Technique Blood collection Serum collection Clean room Washing solution Extra cautions A1 pg L 1 Reference... 

Table VI. Growth of Avirulent Strain A and Virulent Strain C in Samples of Spent Serum Collected at Various Times during the Growth of Strain C (2 5)...

Tsuchihashi et al. (1998) have developed a method for identification of VX metabolites in serum collected from a victim of the Osaka VX incident. In the serum sample, both EMPA and 2-(diisopropylamino-ethyl)methyl sulfide were detected. The techniques using GC-MS and GC-MS-MS were applicable to biological samples such as serum. These results provided the first documented identification of the specific metabolites of VX in victims serum and clarified a part of the metabolic pathway of VX in the human body. In addition, methods have been developed for measuring the VX-inhibited AChE hydrolytic product EMPA (Noort et al, 1998 2002). 

The early part of the SWHS looked at the relation of pregnancy outcome to maternal TCDD levels measured in serum collected shortly after the explosion. Ninety-seven pregnancies (10.9%) ended as spontaneous abortions. TCDD was associated with a nonsignificant adjusted decrease in gestational age and a 20-50% nonsignificant increase in the odds of preterm delivery. The exposed population also reported symptoms of immune system and neurological disorders however, studies found no link to dioxin. Increases in some forms of cancer found in the exposed population have suggested a link between dioxin and some cancers. 

For bioassays, serum collected in pyrogen-free tubes is required. After rapid centrifugation, freezing of the plasma (or serum) at 80 °C is recommended for storage. Freezingthawing cycles must be prevented. 

Serum or plasma (with heparin or EDTA added to prevent coagulation) can be used depending on the assay method. Samples should be centrifuged within 1 hour of collection and stored frozen at -20 °C or colder until assay. It is recommended that storage before assay under these conditions not exceed 30 days. Some procedures use dried whole blood or serum collected on filter paper. "... 

Sensitization was ascertained by the measurement of specific IgE to mite, cat, dog, grass, milk, egg and peanut in serum collected at age 5. A mix of S. aureus enterotoxins consisting of SEA, SEC and TSST-1 (SE mix) was used to determine classical SE-specific IgE antibodies. Forty-nine of 510 children (9.6%) had SE mix IgE >0.35kUA/l. Boys were more frequently SE mix sensitized than girls, and atopic children were nearly 4 times more commonly SE mix sensitive than nonatopics (19.2 vs. 5.0%, p < 0.0001). Analyses on the relationship between SE mix sensitization and clinically expressed allergic diseases therefore were conducted among the whole study population and atopic children only. 

Figure 3.1.1-1 Example of a stand-alone primary TDAR protocol in rats. Animals (8-10/sex/group) are treated with test compound (low, mid, and high doses) or vehicle by a clinically relevant route for 28 consecutive days. (A) Rats are given a single intravenous injection of KLH on Day 24, and serum collected on Day 29 for determination of IgM specific for KLH. (B) Alternatively, animals are immunized with KLH on Day 15, and serum collected on Days 20 and 29 for measurement of anti-KLH IgM and IgG, respectively.

Biological monitoring is mainly carried out for acute exposures in the workplace, and most typically Involves urine and serum collection. Although the NI content of these fluids does not in general Indicate specific health risks, biological tolerance values of 30 g Ni/L in urine and 8/specific sources of excessive exposure. 

Figure 4, Development of affinity-purified rabbit mti-y-H2AX antibodies. Rabbits were immunized, sera were collected, and immunostaining was performed using nonirradiated and irradiated HeLa cells as described in Materials and Methods. (A) Raw serum collected at 51 or 86 days postimmunization, as indicated. (B) Panels labeled TocV show immunostaining with antibodies obtained by y-H2AXphosphopeptide affinity chromatography of serum from 5 Id bleed of rabbit 1663. Cells were treated with 300 cGy or no radiation as indicated. Phosphopeptide competitor (1 pg/ml) was present where indicated. Panels labeled DAPI" show counterstainingfor DNA.

The total and ionic maternal serum calcium levels of 54 women with normal uncomplicated pregnancies were each significantly less than in normal cord serum collected from several umbilical cords and prior to placental separation. This is accounted for in terms of simple diffusion rather than increased protein binding [121]. The calcium ion-selective electrode has also established the existence of hypocalcaemia during normal uncomplicated pregnancies [122], the ionic serum calcium being 1.11 0.03,1.10 0.02 and 1.05 0.01 mmol dm", respectively, during the first, second, and third trimesters. Hypocalcaemia has also been studied in relation to recurrent apnea in premature infants [124]. 

Direct methodological interferences. Serum collected from patients with renal failure has increased concentrations of compoimds such as urea while high levels of bilirubin are foimd in specimens from subjects with liver dysfunction. These are known to interfere (positively and negatively) with many analytical methods. Lipemic specimens cause light scattering that may produce aberrant results in spectrometric assays. 

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