Direct assessment of international consistency of standards for ground-level ozone: strategy and implementation toward metrological traceability network in Asia.

An international exercise to directly assess consistency of standards for ground-level ozone in East Asia was conducted as part of the East Asian Regional Experiment 2005 (EAREX 2005) in the framework of the Atmospheric Brown Clouds (ABC) project. Ten organizations collaboratively participated in the intercomparison. Four groups representing Japan, Korea, Hong Kong, and Taiwan made comparisons at the Gosan super observatory, Jeju Island, Korea, in March 2005, with ozone instruments calibrated to their national standards, and four Japanese groups made off-site comparisons with laboratory-level standards. All comparisons generally indicated good agreement with the standard reference photometer (SRP) 35, built by the National Institute of Standards and Technology (USA) and maintained by the National Institute for Environmental Studies (Japan). The assessment was expanded to measurement networks contributing to the World Meteorological Organization's Global Atmospheric Watch (WMO/GAW) program as part of off-site comparisons, and excellent agreement was achieved. These efforts contribute to propagating traceability of the national metrology standards among the atmospheric science community, to ensuring comparability of the existing ozone measurements, and to establishing an integrated network of air quality monitoring in Asia.

Influence of solution composition and column aging on the reduction of nitroaromatic compounds by zero-valent iron.

Granular iron is used in reactive permeable barriers for the reductive treatment of organic and inorganic groundwater contaminants. The technology is well established, however, its long-term performance and the importance of the groundwater composition are not yet well understood. Here, the influence of chloride, nitrate, silicate, and Aldrich humic acid on the reactivity of Master Builder iron was studied under anoxic conditions using small packed columns and 2-nitrotoluene (2-NT) as a model contaminant. After initially complete reduction of 2-NT to 2-aminotoluene (2-AT) in the column, possibly under mass-transfer controlled conditions, the reactivity of the iron was found to decrease substantially. In the presence of chloride, this decrease was slowed while exposure to silicate resulted in a very quick loss of iron reactivity. Nitrate was found to interfere strongly with the effect of chloride. These observations are interpreted in terms of corrosion inhibition/promotion and competition. Our results suggest that reactive barrier performance may be strongly affected by the composition of the treated groundwater.

Blocking enzyme-linked immunosorbent assay for detection of antibodies against Actinobacillus pleuropneumoniae serotype 6 in pig serum.

A blocking enzyme-linked immunosorbent assay (ELISA) detecting antibodies against Actinobacillus pleuropneumoniae (Ap) serotype 6 was developed. The blocking ELISA was based on the inhibition of a polyclonal antibody raised against Ap serotype 6. Purified lipopolysaccharide from Ap serotype 6 was used as antigen. The blocking ELISA was tested against sera from pigs experimentally infected with the 12 serotypes of Ap biotype 1. Cross-reaction with serotypes 3 and 8 but not with other serotypes was observed. The sensitivity and specificity of the test on a herd level were evaluated with sera from herds naturally infected with serotypes 2, 6, 8 or 12 and with sera from herds free of infection with any Ap serotype. The blocking ELISA showed a high herd sensitivity (1.00 (0.79-1.00)) and specificity (0.97 (0.93-0.99)).

History and physical examination to estimate the risk of ectopic pregnancy: validation of a clinical prediction model.

STUDY OBJECTIVE: To prospectively validate a clinical prediction model for ectopic pregnancy (EP). METHODS: Prospective cohort with 14-month derivation and 12-month validation phases. All hemodynamically stable, first-trimester patients with abdominal pain or vaginal bleeding who presented to a military teaching hospital emergency department underwent follow-up until an outcome of intrauterine pregnancy (IUP) or EP was established. Patients were separated into the high-risk group, defined as having either peritoneal signs or definite cervical motion tenderness; intermediate-risk group, defined as the presence of pain or tenderness, other than midline cramping, plus absence of fetal heart tones, and absence of tissue visible at the cervical os; and low-risk group (neither high- nor intermediate-risk) using recursive partitioning. RESULTS: Summarizing both phases, 915 patients had 845 (93%) IUPs and 70 (7.6%) EPs, with 18 (1.9%) lost to follow-up. The clinical prediction model classified 75 (8.2%) into the high-risk group (sensitivity 31%, 95% confidence interval [CI] 21% to 44%; specificity 94%, 95% CI 92% to 95%); and 644 (70%) in the intermediate-risk group (sensitivity 98%, 95% CI 89% to 100%; specificity 25%, 95% CI 22% to 29%). The remaining 196 (21%) patients who met neither high-risk nor intermediate-risk criteria were classified into the low-risk group. On the basis of EP prevalence of 7.7%, the risk of EP was less than 1% (95% CI 0% to 3%) for the low-risk group, 7% (95% CI 5% to 10%) for the intermediate-risk group, and 29% (95% CI 19% to 41%) for the high-risk group. CONCLUSION: This clinical prediction model is useful for estimating the risk of EP in first-trimester patients, particularly when ancillary testing is equivocal or not readily available.

Low amniotic fluid volume is poorly identified in singleton and twin pregnancies using the 2 x 2 cm pocket technique of the biophysical profile.

BACKGROUND: This study was done to determine the accuracy of the 2 x 2 cm pocket identifying low amniotic fluid (AF) volume in singleton and twin pregnancies. METHODS: The AF volume was evaluated by ultrasonography for the presence of a 2 x 2 cm pocket before amniocentesis. The actual AF volume was then determined by a diazo-dye reaction with subsequent spectrophotometric analysis using paraminohippurate. RESULTS: The AF volume was low in 21 of the 79 singleton pregnancies and normal in 47; hydramnios was present in 11. Among amniotic sacs of the 60 twin pairs, oligohydramnios was found in 33 amniotic sacs. normal AF volume in 80 sacs, and high volume in 7. An AF pocket smaller than 2 x 2 cm was identified in only 3 of the 79 singleton pregnancies and in only 2 of the 120 twin amniotic sacs. CONCLUSIONS: Judging AF volume on the basis of a 2 x 2 cm pocket misses more than 90% of cases of oligohydramnios in singletons and twins.

Risk factors for secondary arrest of labor among women >41 weeks' gestation with an unfavorable cervix undergoing membrane sweeping for cervical ripening.

OBJECTIVE: To identify the risk factor(s) for secondary arrest of labor among women with an unfavorable cervix at > 41 weeks' gestation. METHODS: Prospectively all gravid women with a Bishop score of < or = 4 and no contraindication to a vaginal delivery were candidates for this study. Univariate analysis followed by logistic regression modeling were used to identify variables with a significant association. RESULTS: Over a 9-month period, 115 women entered into the study. In univariate analysis, variables with a significant association with cesarean delivery: (1) non-Caucasian race (P = 0.007), Bishop score < 7 at hospitalization; P = 0.001, and reason for admission (P = 0.017). Logistic regression analysis yielded OR 4.7 (1.6, 15) non-Caucasian race and 9.5 (3.2, 30.8) Bishop < 7 on admission. CONCLUSION: Pregnancies > 41 weeks with an unfavorable cervix, non-Caucasian race and a failure to achieve a Bishop score of > or = 7 prior to hospitalization were significant risk factors for abdominal delivery.

Derivation of a clinical prediction model for the emergency department diagnosis of ectopic pregnancy.

OBJECTIVE: To derive a clinical prediction model for estimating the pretest probability of ectopic pregnancy in ED patients with first-trimester abdominal pain or vaginal bleeding. METHODS: All hemodynamically stable first-trimester patients presenting to the ED of a tertiary care military teaching hospital over a 14-month period with a chief complaint of abdominal pain and/or vaginal bleeding had clinical data coded prior to determining outcome. They were then followed longitudinally until a criterion standard pregnancy outcome was established. RESULTS: Of the 486 patients enrolled, 280 (58%) had viable intrauterine pregnancies, 167 (34%) had nonviable intrauterine pregnancies, and 39 (8%) had ectopic pregnancies. Using a recursive partitioning model, a high-risk group was derived (that was separated from intermediate and low-risk groups), consisting of patients with abdominal peritoneal signs or definite cervical motion tenderness, with a sensitivity of 31% (95% CI: 17-48%), a specificity of 93% (95% CI: 90-95%), a positive likelihood ratio of 4.3, and a negative likelihood ratio of 0.74. A low-risk group, consisting of patients with either fetal heart tones or tissue at the cervical os, or the absence of pain other than midline menstrual-like cramping and lacking any pelvic tenderness, was differentiated from an intermediate-risk group, with a sensitivity of 96% (95% CI: 81-100%), a specificity of 22% (95% CI: 18-26%), a positive likelihood ratio of 1.2, and a negative likelihood ratio of 0.17. CONCLUSION: A clinical prediction model for estimating the probability of ectopic pregnancy in ED patients has been derived. It may prove to have practical clinical application for estimating pretest probability of ectopic pregnancy as well as assisting in medical decision making when laboratory and ultrasonographic findings are nondiagnostic. Clinical application should await prospective validation in an independent sample.

Do multiple measurements employing different ultrasonic techniques improve the accuracy of amniotic fluid volume assessment?

This investigation was undertaken to determine if the accuracy of the ultrasound assessment of abnormal amniotic fluid volume (oligohydramnios or polyhydramnios) is improved by employing multiple sonographic amniotic fluid measurements. Four ultrasound techniques consisting of the subjective assessment (ultrasonic visualization without measurement), largest vertical pocket, amniotic fluid index and 2-diameter pocket technique were performed followed by amniocentesis and dye-dilution confirmation of amniotic fluid volume in 66 singleton pregnancies. The ultrasound accuracy to detect abnormal amniotic fluid volume ranged from 61% with the largest vertical pocket to 70% with the 2-diameter pocket procedure used separately. Receiver operator characteristic curves demonstrated that combining the 4 ultrasonic measurements did not improve the accuracy of identifying amniotic fluid volumes.

The porcine acute phase response to infection with Actinobacillus pleuropneumoniae. Haptoglobin, C-reactive protein, major acute phase protein and serum amyloid A protein are sensitive indicators of infection.

In an experimental infection model mimicking acute Actinobacillus pleuropneumoniae (Ap) infection in swine (Sus scrofa) by aerosol inoculation, the development of a number of typical clinical signs was accompanied by a prototypic acute phase reaction encompassing fever and an acute phase protein response peaking at around 2 days after infection. Haptoglobin, C-reactive protein (CRP), and major acute phase protein (MAP) responded with large increases in serum levels, preceding the development of specific antibodies by 4-5 days. Serum amyloid A protein (SAA) was also strongly induced. The increase, kinetics of induction and normalization were different between these proteins. It is concluded that experimental Ap-infection by the aerosol route induces a typical acute phase reaction in the pig, and that pig Hp, CRP, MAP, and SAA are major acute phase reactants. These findings indicate the possibility of using one or more of these reactants for the nonspecific surveillance of pig health status.

The accuracy of the summated amniotic fluid index in evaluating amniotic fluid volume in twin pregnancies.

OBJECTIVE: Our purpose was to determine the accuracy of the summated amniotic fluid index designed to estimate the total amniotic fluid volume in twin pregnancies. STUDY DESIGN: The summated amniotic fluid index was measured in 62 normal diamniotic twin pregnancies by adding the deepest vertical pockets in the four quadrants. Actual amniotic fluid volume was then determined in all 124 amniotic sacs by amniocentesis and a dye-dilution technique. For data analysis, amniotic fluid volumes were classified by percentile with use of previously reported norms. RESULTS: There were significant differences in the percentile distribution of amniotic fluid volume as estimated by the summated amniotic fluid index and the actual volume as determined by dye dilution (p < 0.001). The summated amniotic fluid index has a sensitivity of only 13% in predicting amniotic sac volume. CONCLUSION: The summated amniotic fluid index is a poor predictor of intertwin differences in amniotic fluid volume and cannot identify twin pairs at risk for oligohydramnios and hydramnios.

Immunological detection of sheep experimentally infected with strains of Mycobacterium avium subspecies containing insertion sequence IS901/IS902 and a 40 kDa protein.

A monoclonal antibody raised against a 40 kDa protein present in certain M. avium strains (IS901/IS902 positive) was used for developing a blocking ELISA. Sera from experimentally infected sheep were evaluated by indirect ELISA, AGID and blocking ELISA. The blocking assay proved to be highly specific for differentiation of sheep infected with different subspecies of M. avium.

Distribution of serotypes, IS901 and a 40 kDa protein in Mycobacterium avium complex strains isolated from man and animals in Denmark.

The aim of the present study was to characterize all strains of the Mycobacterium avium complex isolated in Denmark in 1993. A total of 141 M. avium complex strains (86 from man, 38 from animals, and 17 from peat) were analysed by serotyping, ELISA specific for a 40 kDa protein, and IS901-specific PCR. Serotype analysis showed that the most frequent serotypes among human strains were serotype 4 (27%) and serotype 6 (19%), which differs from an earlier survey where serotype 1 was most prevalent. The most frequent serotypes in animals were serotype 2 (53%) and serotype 6 (13%), whereas the most prevalent serotypes among strains isolated from peat were serotype 4 (29%) and serotype 9 (18%). There was a concurrent appearance of IS901 and p40 in all strains. Only M. avium complex strains isolated from animals, and belonging to serotype 1 or serotype 2, contained the IS901/p40 markers. The different distribution of serotypes of M. avium complex strains in animals and man, and the presence of IS901/p40 exclusively in animal strains, suggests that transmission of M. avium from animals to man is not of significance in Denmark.

Primary structure of the plant serpin BSZ7 having the capacity of chymotrypsin inhibition.

The primary structure of barley grain serpin BSZ7 was deduced from a cDNA encoding 397 amino-acid residues. More than 70% of the residues were confirmed by sequencing peptide fragments. The N-terminus was identified as an acetylated Ala by using mass spectrometry coupled with amino-acid analysis. None of the four putative N-glycosylation sites were found to be glycosylated. The positional identity of BSZ7 with plant and mammalian serpins is 69-72% and 25-32%, respectively.

Accuracy of ultrasonography in evaluating amniotic fluid volume at less than 24 weeks' gestation.

The purpose of this investigation was to evaluate the accuracy of common sonographic techniques in assessing the amniotic fluid volume in pregnancies of less than 24 weeks' gestation. Patients at less than 24 weeks' gestation undergoing an amniocentesis for the placement of prostaglandin F2 alpha for termination (because of genetic or fetal anomalies, or both) were assessed for amniotic fluid volume. All fetuses were alive at the time of prostaglandin instillation. The amniotic fluid index and two-diameter pocket were used to determine the amniotic fluid volume. Prior to the prostaglandin instillation, the amniotic fluid volume was determined with para-aminohippurate using a diazo dye reaction with spectrophotometric analysis. The amniotic fluid volume was determined in 21 pregnancies between 15 and 24 weeks' gestation, yielding volumes ranging from 189 to 1840 ml. Using published standards for amniotic fluid volume in singleton pregnancies, oligohydramnios was present in three gestations, the volume was found to be normal in 15, and hydramnios complicated three pregnancies. The two-diameter pocket identified the amniotic fluid volumes correctly more often (18 of 21 [85.7%]) than the amniotic fluid index (10 of 21 [47.6%]) (P = 0.02). Normal amniotic fluid volume was identified in nine of 15 (60%) pregnancies by the amniotic fluid index and in 14 of 15 (93.3%) by the two-diameter pocket (P = not significant). Abnormal amniotic fluid volumes, oligohydramnios, and hydramnios were recognized more often by the two-diameter pocket (66.7%) than by the amniotic fluid index (1 of 6 [16.7%], P = not significant).

Two markers, IS901-IS902 and p40, identified by PCR and by using monoclonal antibodies in Mycobacterium avium strains.

The occurrence of two markers, a newly identified 40-kDa protein (p40) and the insertion sequence IS901-IS902, in strains of Mycobacterium avium subspp. was evaluated. Analysis of 184 type and field strains of the M. avium complex from human, animal, and environmental sources by PCR specific to IS901 and by a monoclonal antibody specific to p40 demonstrated the presence of the two molecular markers in all of the M. avium subsp. silvaticum strains examined and also in a number of M. avium subsp. avium strains (the latter isolated mainly from pigs). The appearance of the two markers was completely concurrent in all strains. Further, the marker-positive M. avium subsp. avium strains were mainly serotype 2, whereas M. avium complex strains of serotypes 4, 6, 8, 9, and 10 were marker negative. The M. avium subsp. avium type strains ATCC 25291 and approximately 50% of the M. avium subsp. avium field strains isolated from animals contained the markers, while only one strain of human origin was found to be marker positive. Therefore, IS901 and p40 appear to have substantial potential to differentiate among isolates of the M. avium complex. This observation raises new issues regarding classification of strains, since the presence of the markers was found to be inconsistent with the present taxonomic grouping of M. avium subspp.

Characterization of purified protein derivative of tuberculin by use of monoclonal antibodies: isolation of a delayed-type hypersensitivity reactive component from M. tuberculosis culture filtrate.

Nine monoclonal antibodies were raised against purified protein derivative (PPD) of tuberculin in mice previously treated with Bacilli Calmette Guérin (BCG). The antibodies also reacted with a culture filtrate from Mycobacterium tuberculosis strain H37Rv. In immunoblotting after SDS-PAGE the reaction with PPD was seen as a diffuse smear, whereas ammonium sulphate-precipitated proteins from H37Rv gave well-defined bands ranging from 10 to 65 kDa. Enzyme immunoassay showed that both PPD and H37Rv antigens were able to inhibit binding of the antibodies to PPD coated microtitre wells, suggesting that the antibodies reacted with continuous epitopes. A 12 kDa protein purified by immunoaffinity chromatography from H37Rv antigens was tested intradermally in M. tuberculosis MNC3 sensitized guinea pigs and gave a delayed type hypersensitivity reaction.

Identification and characterization of a putative serine protease expressed in vivo by Mycobacterium avium subsp. paratuberculosis.

A putative serine protease expressed in vivo by Mycobacterium avium subsp. paratuberculosis was isolated from a lambda gt11 genomic expression library by screening with serum from a naturally infected sheep. The gene was contained in two overlapping clones, which were shown by antibody elution to encode a protein of 34 kDa in M. a. paratuberculosis. The clones were sequenced and database searches detected a motif identical to the active serine site in trypsin, and 30% homology to the putative serine proteases (HtrA proteins) of Escherichia coli, Salmonella typhimurium, Brucella abortus and Rochalimaea henselae.

Linkage between serum cholinesterase 2 (CHE2) and gamma-crystallin gene cluster (CRYG): assignment to chromosome 2.

Serum cholinesterase 2 (CHE2) was examined in a Danish material of normal families that has been tested earlier for 70-78 classical marker systems and 25 RFLP systems. DNA for RFLP typing was provided by transforming 16-year-old frozen lymphocytes. The frequency of allele CHE2*C5+ in the Danish population was found to be 0.0430. The highest lod score was between CHE2 and the gamma-crystallin gene cluster (CRYG) (zeta = 4.21 at theta = 0.00 in females). The scores were from a single family with 15 children. CHE2 may, accordingly, be assigned to the location of CRYG: chromosome 2, bands q33-q35.