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The development of the eye lens in the prenatal phase is accompanied by a gradual change in the polypeptide composition of α-crystallin. As the lens matures from that of a 6-week-old embryo to that of a new-born calf, there is a successive appearance of different polypeptides although two obviously principal polypeptides, A2 and B2, are present at every stage of development. The youngest embryonic lenses studied contain four polypeptides. Two of them differ from the well-known polypeptides as found in cortical bovine α-crystallin.

Background Decreased amyloid β (1-42) (Aβ42) and increased (phosphorylated) tau in cerebrospinal fluid (CSF) are considered to be a reflection of plaques, tangles, and neuronal degeneration in Alzheimer's disease (AD). Atrophy of the medial temporal lobe (MTA) on magnetic resonance imaging (MRI) reflects neuronal loss in this area. Objective To compare diagnostic accuracy of CSF biomarkers and MTA in AD versus controls. Methods Aβ42, tau and tau phosphorylated at threonine 181 (Ptau-181) were measured in CSF from 61 AD patients and 32 controls by sandwich enzyme-linked immunosorbent assay. A CSF biomarker profile for AD was constructed. MTA was rated visually on MRI. Results When AD patients and controls were evaluated separately, no correlations were present between the CSF markers and MTA score. Both MTA and CSF biomarker profile were independently associated with the diagnosis AD (MTA: OR (95% CI) = 28 (3–239); CSF biomarker profile: OR (95% CI) = 57 (13–262)). Among individuals younger than 65 years old and without MTA 60% suffered AD, and the finding of an abnormal CSF biomarker profile was limited to AD patients only. Conclusions MTA and CSF biomarkers seem to be of incremental value for the diagnosis AD. CSF analysis is most sensitive in the absence of MTA, and especially among early-onset AD patients.

Various C-terminally truncated amyloid beta peptides (Abeta) are linked to Alzheimer's disease (AD) pathogenesis. Cerebrospinal fluid (CSF) concentrations of Abeta38, Abeta40, and Abeta42 were measured by enzyme-linked immunosorbent assay in 30 patients with AD and 26 control subjects. CSF Abeta42 levels was decreased in patients with AD, whereas CSF Abeta38 and Abeta40 levels were similar in patients with AD and control subjects. All three Abeta peptides were interrelated, particularly CSF Abeta38 and Abeta40. Diagnostic accuracy of CSF Abeta42 concentrations was not improved by applying the ratios of CSF Abeta42 to Abeta38 or Abeta40.

During cardiopulmonary bypass (CPB), the brain and the kidneys may be damaged because of microemboli, ischemia, and inflammation. The latter has been reduced by the use of heparin coated circuits. We questioned whether heparin coated circuits could also reduce cerebral and renal damage and whether inflammatory markers correlate with damage to the brain and the kidneys. Fifty-one patients scheduled for coronary artery bypass grafting were perfused with either a heparin coated or an uncoated circuit. To compare the effect of a heparin coated circuit with an uncoated circuit upon cerebral and renal function in relation to inflammation, we assessed markers of cerebral (S100beta) and renal (N-acetyl-beta-D-glucosaminidase [NAG], creatinine, and urea) function, inflammation, and oxygen metabolism. S100beta levels and NAG levels increased during CPB in both groups as compared with baseline levels (p

Background: Reported concentrations of amyloid β (1–42) (Aβ42) and tau in cerebrospinal fluid (CSF) differ among reports. We investigated the effects of storage temperature, repeated freeze/thaw cycles, and centrifugation on the concentrations of Aβ42 and tau in CSF. Methods: Stability of samples stored at −80 °C was determined by use of an accelerated stability testing protocol according to the Arrhenius equation. Aβ42 and tau concentrations were measured in CSF samples stored at 4, 18, 37, and −80 °C. Relative CSF concentrations (%) of the biomarkers after one freeze/thaw cycle were compared with those after two, three, four, five, and six freeze/thaw cycles. In addition, relative Aβ42 and tau concentrations in samples not centrifuged were compared with samples centrifuged after 1, 4, 48, and 72 h. Results: Aβ42 and tau concentrations were stable in CSF when stored for a long period at −80 °C. CSF Aβ42 decreased by 20% during the first 2 days at 4, 18, and 37 °C compared with −80 °C. CSF tau decreased after storage for 12 days at 37 °C. After three freeze/thaw cycles, CSF Aβ42 decreased 20%. CSF tau was stable during six freeze/thaw cycles. Centrifugation did not influence the biomarker concentrations. Conclusions: Repeated freeze/thaw cycles and storage at 4, 18, and 37 °C influence the quantitative result of the Aβ42 test. Preferably, samples should be stored at −80 °C immediately after collection.

Background: In this study the clinical and technical performance of the CA125- detecting Bayer ACS:OV immunoluminometric serum assay was compared with three other well-established CA125-detecting assays. Methods: A total of 1112 serum samples was included in this evaluation: 462 from apparently healthy women, 153 from patients with benign ovarian disease, 163 from patients with malignant ovarian disease, 10 from patients with borderline ovarian malignancies and 78 samples from 12 ovarian cancer patients during monitoring of disease. Serum samples from women with malignant endometrial ( n = 68) and colon ( n = 32) diseases were also included. Moreover, serum samples from women with benign uterine disease and endometriosis ( n = 136) plus 10 serum samples from men ( n = 7) and women ( n = 3) with human anti-mouse antibodies (HAMA) after immunoscintigraphy were included. All samples were tested in duplicate with the Bayer ACS:OV, the Centocor CA125 II, the Abbott IMx CA125 and the Roche (formerly Boehringer Mannheim) Enzymun-Test® CA125 II assays. Results: The clinical performance of the Bayer ACS:OV assay, assessed in various patient groups, was similar to that of the two other automated assays. In serum from patients with benign diseases the highest values were found in patients with benign ovarian tumours. In the ovarian cancer patients followed during the course of disease we found similar marker patterns with all four assays. In contrast to the Roche Enzymun-CA125 II assay and to a lesser extent the Centocor CA125 II assay, the Bayer ACS:OV assay was less sensitive to interference from HAMA. Conclusion: The Bayer ACS:OV assay is a precise and reliable test for the quantification of CA125 in serum.

The purpose of this study was to assess the differential diagnostic potential of a combination of CA 125, CA 15-3, and CA 72-4 antigens in the definition of malignant disease, especially ovarian carcinoma in patients with a pelvic mass.A total of 412 patients were evaluated in a multicenter, retrospective study.Two hundred twenty-six malignant, 171 benign pelvic tumors (of which 129 were benign ovarian tumors), and 15 borderline tumors were evaluated. One hundred thirty-three patients had ovarian carcinoma. In 76 cases (55%), the International Federation of Gynecology and Obstetrics stage was III or IV. Borderline tumors (n = 15) were excluded from the statistical calculations. CA 125 antigen was the most sensitive marker for ovarian carcinoma (81%). The highest specificity and positive predictive value was obtained with CA 15-3 antigen (95% and 92%, respectively). Considering a concomitant elevation of all 3 markers as positive, a positive predictive value of 97% was found. However, only 28% of the patients in the total group and 41% of the patients with ovarian carcinoma had a concomitant elevation of all 3 markers. The combination of all 3 markers with levels below the cut-off resulted in a (false-positive) positive predictive value for malignancy between 12% and 36%. With the use of logistic regression analysis, we found a correct prediction in 73% of the cases. CA 15-3 antigen makes the most significant (P.0001) contribution to the logistic model in the prediction of malignancy in the total group, with all pelvic masses with an odds ratio of 3.86.The combination of a simultaneous elevated level of CA 125, CA 15-3, and CA 72-4 antigens was predictive for malignant disease in almost all cases. However, such concomitant elevation was found in few of the malignant masses. Logistic regression analysis revealed that CA 15-3 antigen makes the most significant contribution to a model for the prediction of malignancy in the total group. The logistic model gave a correct prediction in 73% to 83%. The present tumor marker panel seems inferior to combinations with other test modalities, which include ultrasonography and/or physical examination and/or menopausal status or age.

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder of unknown aetiology, characterized by irreversible cognitive and physical deterioration. It is a major cause of morbidity and death in the elderly and a growing public health problem as life expectancy in the general population increases. AD is both genetically and phenotypically a heterogeneous disorder. An early-onset, familial type is recognized, as well as a later-onset, sporadic type. The diagnosis is made on clinical grounds, with the aid of a small number of additional investigations, using consensus criteria. 1 However, at autopsy about 10± 20% of clinically diagnosed AD patients are found to have conditions other than AD. Therefore, genetic and/or biochemical markers that support the clinical diagnosis and can distinguish AD from cognitive symptoms attributable to ageing and from other dementias will be of great value. The identi® cation of such accurate markers for the early diagnosis of AD is mandatory for the development of ef® cient pharmacological treatment, since therapy should be initiated at an early stage of the disease, before extensive and irreversible brain damage has occurred.

Antibodies (Abs) to MUC1 occur naturally in both healthy subjects and cancer patients and can be induced by MUC1 peptide vaccination. We compared the specificity of natural and induced MUC1 Abs with the objective of defining an effective MUC1 vaccine for active immunotherapy of adenocarcinoma patients. Serum samples, selected out of a screened population of 492 subjects for their high levels of IgG and/or IgM MUC1 Abs, were obtained from 55 control subjects and from 26 breast cancer patients before primary treatment, as well as from 19 breast cancer patients immunized with MUC1 peptides coupled to keyhole limpet hemocyanin (KLH) and mixed with QS-21. The samples were tested with enzyme-linked immunoassays for reactivity with (1) overlapping hepta- and 20-mer peptides spanning the MUC1 tandem repeat sequence; (2) two modified 60-mer peptides with substitutions in the PDTR (PDTA) or in the STAPPA (STAAAA) sequence of each tandem repeat; and (3) four 60-mer glycopeptides with each 1, 2, 3 and 5 mol N-acetylgalactosamine (GalNAc) per repeat. More than one minimal epitopic sequence could be defined, indicating that Abs directed to more than one region of the MUC1 peptide core can coexist in one and the same subject. The most frequent minimal epitopic sequence of natural MUC1 IgG and IgM Abs was RPAPGS, followed by PPAHGVT and PDTRP. MUC1 peptide vaccination induced high titers of IgM and IgG Abs predominantly directed, respectively, to the PDTRPAP and the STAPPAHGV sequences of the tandem repeat. Natural MUC1 Abs from breast cancer patients reacted more strongly with the N-acetylgalactosamine (GalNAc) peptides than with the naked 60-mer peptide, while reactivity with the GalNAc-peptides was significantly reduced (2-tailed p < 0.0001) in the MUC1 IgG and IgM Abs induced by MUC1 peptide vaccination. Whereas in cancer patients glycans appear to participate in epitope conformation, the epitope(s) recognized by MUC1 Abs induced by peptide vaccination are already masked by minimal glycosylation. Therefore, our results indicate that a MUC1 glycopeptide would be a better vaccine than a naked peptide. Int. J. Cancer 86:702–712, 2000. © 2000 Wiley-Liss, Inc.

Objective : To compare the performance of four serum assays for the quantification of MUC1 in breast cancer patients. Study design : A total of 282 serum samples were evaluated with two automated (Boehringer Mannheim Enzymun-Test® CA 15-3 and Chiron ACS™ BR) and two manual assays (Centocor CA 15-3® radioimmunoassay [RIA] and Biomira Truquant® BR RIA™). Sera were obtained from healthy controls ( n =50), patients with benign ( n =25) and malignant breast disease ( n =77) and patients with other malignancies ( n =69). In addition, sera from pregnant women ( n =56) and patients with liver cirrhosis ( n =5) were included. Results : Intraassay coefficients of variation (C.V.s) were highest for the manual Centocor CA 15-3 assay (7.4% for values below 50 kU/l and 8.1% for values above 180 kU/l). Interassay C.V.s were highest for the manual Truquant BR assay (11.7% for the lower concentration values and 18.6% for the higher concentration values). False positive rates ranged between 0% for the Centocor CA 15-3 RIA and 14% for the ACS BR assay (cut-off: 30 kU/l). In monitoring breast cancer patients all four assays show similar patterns, although absolute MUC1 values found may differ up to 50%. Conclusion : For monitoring purposes all assays perform equally well, however, automated assays show lower inter- and intraassay variability, especially in the higher value range. Therefore we recommend the use of the same, automated, assay for quantification of MUC1 during the follow-up of breast cancer patients.

Seven CA 125 immunoassays were compared for their clinical performance. CA 125 concentrations were determined in 289 serum samples obtained from women with benign pelvic tumors (samples from 98 patients) and patients with various cancers (samples from 111 patients). In the range of 0–1000 kilounits/L, all assays tested were linearly correlated, with correlation coefficients ranging from 0.89 to 0.99. In relation to the original Centocor CA 125 assay, there was an overall tendency to measure higher absolute values in the lower CA 125 value range. This was not seen in relation to the Centocor CA 125 II assay. ROC curves (benign vs pretreatment ovarian cancer patients) were nearly identical for all assays, and the areas under the ROC curves were not markedly different. We conclude that the CA 125 assays tested are strongly related to each other and are clinically reliable for the quantification of serum CA 125 and that none of the assays offers higher diagnostic accuracy or better discrimination between patient groups, especially not in the lower ranges.

The mucin glycoprotein-detecting assay CA 15-3 is a valuable tool for monitoring the course of disease in breast cancer patients. Assays of CA 15-3 are based on the use of two MAbs to polymorphic epithelial mucin (PEM). We evaluated the technical and clinical performance of the Chiron ACSTM BR, an automated competitive chemiluminescence assay using a single MAb, B27.29, and compared the assay’s results with those of the Centocor CA 15-3 RIA, the Abbott IMx CA 15-3, and the Boehringer Mannheim Enzymun-Test CA 15-3. The study population consisted of 253 healthy women, 66 patients with benign breast disease, 168 breast cancer patients, and 76 patients with other carcinomas. In the technical evaluation, we assessed the precision and linearity on dilution of the ACS BR assay. Cutoff values (upper limits of values seen in healthy subjects) were determined for all four assays. Agreement between the assays was studied by linear regression analysis. The ACS BR assay gave within- and between-assay CVs of 2.2% and 3.9%, respectively. Three samples from healthy women gave discordant values by ACS BR and were not included in the calculations. All four assays exhibit a highly similar pattern when monitoring breast cancer disease; the closest agreement of values was obtained between ACS BR and Centocor CA 15-3. We conclude that the ACS BR assay is a fast and reliable immunoassay for measuring PEM in serum. Although it detects a slightly different epitope on the PEM molecule than is targeted in other assays, for cancer serum samples it agreed better with the original Centocor CA 15-3 assay than did the other two CA 15-3 assays tested.

The Byk LIA-mat CA125 II assay was compared with the Centocor IRMA CA125 II. Serum samples studied (n = 1012) were obtained from 652 apparently healthy females, 61 pregnant women, and 299 patients with benign and malignant gynecological tumors. The CA125 II assay value at the 95th percentile of the total healthy group was 29 kU/L for the LIA-mat and 32 kU/L for the Centocor assay. For the LIA-mat assay the 95th percentile was 31 kU/L (Centocor 36 kU/L) for the group 55 years of age. By using ROC curves we found the optimal pretreatment Byk LIA-mat CA125 II value differentiating between benign and malignant ovarian tumors to be 95 kU/L. Pretreatment CA125 values >1000 kU/L were detected in serum samples of patients with advanced epithelial ovarian cancer.

In this report, we evaluate the stability of urinary gonadotropin peptide (UGP), an urinary tumor marker for trophoblastic malignancies, which is also promising for other types of cancer. The concentrations were independent of the duration of storage (p0.1), but the mean difference between UGP values for samples stored at 4 degrees C and those kept at room temperature is 5.5% (p0.002), with the higher values for refrigerated samples. We conclude that UGP is stable at room temperature, allowing mailing of urine samples. Refrigerated (4 degrees C) samples show slightly increased values. However, this will have no clinical relevance in monitoring cancer patients.

Objective: To compare the lipid and (apo-)lipoprotein profile and blood pressure of men with long-standing spinal cord injuries (SCI) to those of an age-matched able-bodied (AB) population, and to assess the most important determinants of this pro file and blood pressure. Design: A cross-sectional study of persons with chronic SCI residing in the community. Setting: Tests were performed in a university research laboratory. Subjects: Thirty-seven men (age 37.4 ± 12.0yrs) with longstanding (14.7 ± 8.6yrs) SCI ranging from level C4/5 to L5 volunteered to participate. Comparisons were made with published data from 3,498 AB men, age 20 to 59 yrs, from the same country. Main Outcome Measures: Lipid and lipoprotein profile (total cholesterol [TC], high-, low-, and very low density lipoprotein cholesterol [HDL-C, LDL-C and VLDL-C, respectively], and triglycerides [TG]), as well as aerobic power, activity level, anthropometric variables, and blood pressure. Multiple regression analyses assessed the most important determinants of the lipid and blood pressure profile. Results: None of the lipid variables were related to the lesion level. TC, HDL-C, and TC/HDL-C were not significantly different from the AB population. The most important determinants of TC, LDL-C, and the ratios TC/HDL and HDL-C/LDL-C were age, smoking behavior, and activity level. Aerobic power was not an important determinant of any lipid or (apo- )lipoprotein or blood pressure. Conclusion: Men with long-standing SCI do not appear to have an essentially different coronary heart disease risk profile compared with AB persons. Modifiable risk factors such as activity level, smoking, alcohol consumption, body mass index, and adipose tissue were more important than lesion level and aerobic power in the determination of the lipid and lipoprotein profile, suggesting several potential interventions.

Homocysteine accumulation, frequently observed in plasma of AD patients, may be a sign of a reduced activity of the brain methionine-homocysteine transmethylation cycle. S-Adenosylmethionine (SAM) is the main methyl donor in several transmethylation reactions. The demethylated product of SAM, S-adenosylhomocysteine (SAH), is hydrolyzed to yield homocysteine, which can be remethylated to methionine by transfer of a methyl group of 5- methyltetrahydrofolate (5-MTHF). A reduced activity of the transmethylation cycle in the brain may result in hypomethylation of the promoter of the presenilin 1 (PS1) gene, which will lead to overexpression of presenilin 1 and, consequently, to increased Aβ1-42 (Aβ42) formation. Brain transmethylation was studied in 30 patients with 'probable' AD and 28 age-matched non-demented controls by measuring the cerebrospinal fluid (CSF) levels of SAM, SAH and 5-MTHF. 5-MTHF was determined by HPLC with electrochemical detection, while SAM and SAH were assayed by stable isotope dilution tandem mass spectrometry. We found no statistical differences between AD patients and controls for 5-MTHF, SAM and SAH levels, and the SAM/SAH-ratio in CSF. These findings argue against a possible change in methylation of the promoter and expression of PS1.

The human anti-(mouse Ig) antibody (HAMA) response was measured in serum of 52 patients suspected of having ovarian carcinoma who had received an i.v. injection of either the murine monoclonal antibody (mAb) OV-TL 3 F(ab')2 (n = 28, 1 mg) or the chimeric mouse/human mAb MOv18 (cMOv18; n = 24, 3 mg). Serum samples were taken before injection and 2-3 and 6-14 weeks after administration. A double-antigen or bridging assay was developed to detect responses against both murine as well as chimeric antibodies. In addition, an indirect enzyme-linked immunosorbent assay (ELISA) as well as three commercially available assays were used to study antibody response against the murine antibody OV-TL 3. With both the double-antigen (bridging) assay and the indirect ELISA 1 of the 28 patients (4%) injected with murine OV-TL 3 F(ab')2 showed a HAMA reaction 6 weeks after injection, which was demonstrated to be a mixed anti-isotypic and anti-idiotypic response. None of the 24 patients injected with the chimeric MOv18 showed an anti-chimeric antibody response. The various commercially available assays demonstrated conflicting results. The double-antigen- or bridging assay is a reliable method to detect anti-murine and anti-chimeric antibodies. The assay can be easily adapted for use with human antibodies. The immunogenicity of OV-TL 3 F(ab')2 and cMOv18 in patients is low, making both antibodies candidates for immunotherapy.

OBJECTIVE To study whether serially measured plasma concentrations of endothelin (a novel, potent, endogenous vasoconstrictor derived from endothelium and macrophages) relate to the pathophysiology and severity of human septic shock. DESIGN Prospective analysis. SETTING Medical ICU of a university hospital. PATIENTS Six patients with septic shock, studied for 8 days after ICU admission. MEASUREMENTS AND MAIN RESULTS The initial plasma endothelin concentration was increased (14.2 +/- 5.2 [SD] vs. normal 4.2 +/- 0.7 pg/mL, p less than .05) and correlated with the Acute Physiology and Chronic Health Evaluation II score (r2 = .79, p less than .05). For pooled data, endothelin levels correlated poorly with leukocyte counts (r2 = .13), mean arterial pressure (MAP) (r2 = .16), and administered doses of dopamine (r2 = .26). In multiple regression analyses, plasma endothelin concentrations were predicted by dopamine doses and not by MAP. Plasma endothelin concentrations predicted the decrease in creatinine clearance, independently from MAP. The pooled value for correlations between endothelin levels and creatinine clearance, during the course of disease in individual patients, was statistically significant (r2 = .31). CONCLUSIONS During septic shock, the release or production of endothelin may increase as a consequence of endothelial injury by activated leukocytes and the infusion of catecholamines, and this mechanism may relate to renal vasoconstriction and to the severity of disease.

Plasma endothelin levels were studied in 29 pregnancies. Levels were found to be increased in 9 pregnancies complicated by pregnancy-induced hypertensive disease and/or pre-eclampsia as compared to levels in 14 normotensive pregnancies with gestational age varying between 24–40 weeks with normal Doppler flow velocity waveforms of the uterine arcuate arteries, and 6 normotensive pregnancies with abnormal Doppler flow velocity waveforms at 24 weeks' gestation. Two patients with severe pre-eclampsia showed a rise in plasma endothelin levels, suggesting a correlation between plasma endothelin levels and the disease process. Endothelin is produced by endothelial cells of blood vessels; it is the most potent vasoconstrictor known to date. For this reason it has been suggested that endothelin might be important in the control of systemic blood pressure and local blood flow, both disturbed in pre-eclampsia. The conclusion of this study is that the venous plasma level of endothelin would seem to be a marker for severe disease, however, without any predictive value.

Amyloid β 42 (Aβ 42) concentrations in cerebrospinal fluid (CSF) are used to identify Alzheimer disease (AD) (1), but reported concentrations differ among studies, as does diagnostic accuracy (2). These differences may relate to the patient and control groups studied (3), processing and storage methods (4), intra- and interassay variation of the assays, or to the reagent antibodies used. A recent metaanalysis (2) stressed the importance of standardizing assays for Aβ–42 in CSF. In most studies, CSF Aβ 42 was reported to be decreased, but in 2 studies, CSF Aβ 42 was not significantly changed in AD (2), and in 1 study(5) even increased in the early stages of disease. These dissimilarities might reflect the specificities of the antibodies incorporated in the assays. The first aim of our study was to compare Aβ 42 concentrations measured by 2 different assays in the same CSF samples. The first assay, widely used in Europe (6), uses 2 monoclonal antibodies (mAbs) and detects the full-length Aβ 42 peptide, Aβ (1–42) (7). The second assay [Aβ (N–42)], used mainly in the United States (8), detects both full-length Aβ 42 and Aβ peptides truncated at the NH2 terminus (9). The second aim of our study was to compare diagnostic accuracies of the assays for patients with AD compared with controls without dementia and patients with frontotemporal lobar degeneration (FTLD). Finally, we investigated the relationship between CSF Aβ (1–42) and Aβ (N–42) concentrations and albumin ratio, age, disease duration, and disease severity. Between October 2000 and December 2002, we recruited 39 AD patients, 24 FTLD patients, and 30 non-dementia controls at the Alzheimer Center of the VU University Medical Center (VUMC). All patients underwent a standardized investigative battery (3). A diagnosis of “probable” AD was made according to the NINCDS-ADRDA criteria (10); the …

THE apolipoprotein E (ApoE) e4 polymorphism has been associated with increased risk of cardiovascular disease, stroke, Alzheimer’s disease, and impaired cognitive function (1). ApoE e4-associated increases in mortality rates have also been reported, especially at younger ages. Multiple mechanisms may contribute to the effects of ApoE, including its roles in total cholesterol and low density lipoprotein metabolism (2), which are mediated through allele-specific differences in the clearance of different lipoproteins. Given the major disease and mortality rate effects of ApoE e4, significant impacts on functional ability in old age could be expected. Indeed, Gerdes and colleagues (3) have advocated ApoE as a “frailty” gene, yet thus far little evidence of major effects on disability has appeared. Albert and colleagues (4) studied 218 nondemented elderly persons aged 58–93 years, and reported an association between the e4 allele carriers and poorer scores on activities of daily living. However, Bader and colleagues (5) found no association of ApoE genotypes with activities of daily living in 162 Italian octogenarians and nonagenarians. Carmelli and colleagues (6) studied 390 males with a mean age of 72 years and observed a nonsignificant effect of ApoE genotypes on tested lower extremity function. Blazer and colleagues (7) similarly reported that e4 was not associated with incident self-reported functional declines as a main effect, in 1529 participants of mean age 77.8 years, although interactions were found in women. In further work in the Duke Established Populations for Epidemiologic Studies of the Elderly (EPESE) site, Blazer and colleagues (8) also showed that there was no association between e4 status and any of five dimensions of quality of life, and argued that their findings “challenge the uncritical assumption that the presence of this susceptibility gene in the population implies an excess burden of poor quality of life.” Restricted (walking) mobility is an early and relatively culture-free marker of the development of disabilities. In the Women’s Health and Aging Study (9) and the 1984 Supplement on Aging (10), well over 90% of respondents with any disability reported problems with mobility. In addition, there is good evidence that poor performance on gait speed and chair stand tests in older respondents is associated with increased risk of subsequent disability and mortality (11,12). By contrast, questionnaires eliciting self-reported mobility disability inevitably incorporate the effects of environment and attitude (13,14), and are subject to “category response shifts” (15,16). In this study we examined the impact of the ApoE e4 polymorphism on mobility in old age in a large population-based study, using self-report and performance tests, to establish both subjectively and objectively whether e4 is associated with excess mobility impairment in older people.

The aim of the present study was to evaluate the clinical performance of the CA 15-3 assay on Elecsys systems in an international multicenter study (11 centers). A total of 1326 single samples (272 apparently healthy individuals, 34 pregnant women, 308 benign diseases, 273 cancers other than breast, 439 breast cancer) and 538 serial samples of 98 breast cancer patients during follow-up were analyzed. 95% of values in healthy individuals were below 25 kU/L, and 88% in benign breast diseases, respectively. In malignant breast disease at primary diagnosis the value distribution of Elecsys CA 15-3, sensitivity at 95% specificity, as well as the areas under the curve in ROC analysis were clearly correlated to tumor stages: UICC I to IV 88 to 25% of values25 kU/L, sensitivity 7 to 78%, areas under the curve 0.53 to 0.94. During follow-up, sensitivity/specificity for detection of recurrences were 90%/71%. In metastatic disease clinical progression/response to therapy were indicated in 91%/78% of patients at a specificity of 92%/78%. The findings indicate that the Elecsys CA 15-3 assay is very suitable in routine work for detection of recurrences as well as for therapy control in metastatic breast cancer.

Although the sensitivity and specificity of CA 15-3 in primary diagnosis of breast cancer is limited, CA 15-3 measurement has found widespread use in monitoring disease progression and response to therapy [reviewed in Ref. (1)]. A new assay for determination of CA 15-3 on Elecsys® immunoanalyzers has been introduced by Roche Diagnostics GmbH. The analytical performance of this assay was tested in an international multicenter evaluation in 11 evaluation sites, and the clinical performance was judged in 4 of these laboratories. The results are described in this report. Elecsys CA 15-3 is a sandwich immunoassay that uses biotinylated 115D8 and ruthenylated DF3 as capture and detection antibodies, respectively, and electrochemiluminescence as the detection technology. Details of the test principle have been described previously (2). The test is standardized against the Enzymun-Test CA 15-3 (Roche Diagnostics GmbH) in the laboratories of the manufacturer. The analytical evaluation was carried out according to a standardized protocol of the European Committee for Clinical Laboratory Standards (3). The results of within-run imprecision (21 replicates per analysis) and between-day imprecision (1 replicate analyzed on 21 different days) studies are available as a data supplement at Clinical Chemistry Online (http://www.clinchem.org/content/vol47/issue12). The medians of the within-run and between-day CVs were 2.5–3.8% and 5.0–5.9%, respectively, at all concentrations tested (8.4–251.7 kilounits/L) for human sera and control sera. Confirmation of target values assigned to control sera was tested in an interlaboratory survey and was 97% for PreciControl Tumor Marker Level 1 (18.4 kilounits/L) and 96% for Level 2 (58.6 kilounits/L). The medians of all participants of an interlaboratory survey of the German Society for Clinical Chemistry were confirmed for each of the two control samples used (11.8 and 16.8 kilounits/L) by 99% (on the basis of the medians of all participants in the Elecsys study); thereby demonstrating the excellent agreement …

Objective: To analyse whether pregnancies resulting in a small for gestational age neonate are preceded by a prostacyclin deficiency or an imbalance between thromboxane and prostacyclin. Study design: At five fixed time points during pregnancy, 24-h urine samples were collected for the measurement of thromboxane and prostacyclin metabolites thromboxane-B 2 (TXB 2 ) and 6-keto-prostaglandin F 1α (6-keto-PGF 1α ). In order to study trend differences between pregnancies with appropriate (AGA; n =26) and small for gestational age neonates (SGA; n =17), trend analysis with simple contrasts were accomplished for TXB 2 , 6-keto-PGF 1α and the TXB 2 /6-keto-PGF 1α ratio. Results: Trend analysis showed higher TXB 2 levels and higher TXB 2 /6-keto-PGF 1α ratios in patients with SGA versus AGA newborns. No statistically significant difference in 6-keto-PGF 1α excretion between patients with SGA and AGA newborns was detected. Conclusion: The birth of an SGA neonate is not preceded by prostacyclin deficiency. With ongoing pregnancy an imbalance between thromboxane and prostacyclin becomes more obvious in pregnancies with SGA newborns.

Objective: The objective of this study was to evaluate the extent to which endothelin and the eicosanoids prostacyclin and thromboxane A 2 are involved in the pathophysiology of gestational hypertension and preeclampsia. Study Design: In a longitudinal design, venous blood samples and 24-hour urine specimens were collected from 396 women in each trimester of pregnancy. After delivery of all patients, venous plasma endothelin was assessed in 20 subjects with identified preeclampsia, 48 subjects with gestational hypertension, and 59 normotensive subjects. Urinary excretions of the thromboxane A 2 and of the prostacyclin metabolites thromboxane B 2 and 6-keto-prostaglandin F 1 α were assessed in 16 subjects with preeclampsia, 35 subjects with gestational hypertension, and 31 normotensive subjects. Results: Endothelin levels showed a second-trimester drop in all groups. In all 3 gestational trimesters a high correlation was found between the excretion of thromboxane B 2 and that of 6-keto-prostaglandin F 1 α ( P 2 and 6-keto-prostaglandin F 1 α urinary excretions increased throughout pregnancy and the overall thromboxane B 2 /6-keto-prostaglandin F 1 α ratio decreased. No significant differences in endothelin, thromboxane B 2 , and 6-keto-prostaglandin F 1 α excretion levels or in thromboxane B 2 /6-keto-prostaglandin F 1 α ratios were found between women with preeclampsia, gestational hypertension, and normotension. Only in a small group of patients with severe preeclampsia (n = 2) and severe gestational hypertension (n = 2) were increased second-trimester endothelin values and increased thromboxane B 2 /6-keto-prostaglandin F 1 α ratios found. Conclusion: In this longitudinal study we found no evidence for prostacyclin deficiency or increased endothelin levels in preeclampsia. Only women with severe preeclampsia and severe gestational hypertension expressed increased endothelin levels and thromboxane dominance over prostacyclin. (Am J Obstet Gynecol 1998;179:1559-64.)

To assess the usefulness of biliary CEA determinations in the diagnosis of recurrent tumour, gallbladder bile was sampled in patients who underwent laparotomy for proven or suspected recurrent colorectal cancer and in control patients. Biliary CEA concentrations in controls were < 5 ng/ml, whereas significantly elevated CEA concentrations were found in the bile of all patients with tumour recurrence. Serum concentrations in these patients were elevated in 77% only. In a series of 12 patients with (a) suspicious lesion(s) on liver imaging but normal serum CEA concentration during follow-up, biliary CEA determination differentiated clearly between metastases and benign lesions. Biliary CEA determination seems to aid detection of tumour recurrence at an early stage and may preclude unnecessary surgery in patients with undefined liver lesions.

The effect of estrogen on the glycosylation of alpha 1-acid glycoprotein was studied in women using oral contraceptives and in male-to-female transsexuals receiving oral or transdermal estrogen treatment. Oral estrogen treatment induced an increase in degree of branching and a decrease in fucosylation and sialyl Lewis x expression on alpha 1-acid glycoprotein compared to individuals receiving no estrogens or transdermal estrogen treatment. The effect on the glycosylation of alpha 1-acid glycoprotein was less in the male-to-female transsexuals receiving oral estrogens in combination with cyproterone acetate, a blocker of the androgen receptor with progestagen-like effects. This was of comparable magnitude as in women using oral contraceptives containing both estrogen and progestagen. We conclude that oral estrogens have an identical effect on the hepatic glycosylation of alpha 1-acid glycoprotein in both males and females and that progestagen reduces this effect. These results show that oral estrogens induce an effect on the glycosylation of alpha 1-acid glycoprotein opposite to that induced by inflammation. Oral estrogens can therefore modulate the glycosylation dependent inflammatory actions of alpha 1-acid glycoprotein, while this is not the case with transdermal estrogens. In all likelihood, estrogen receptors on the hepatocyte must play a significant role in the observed effect.

OBJECTIVE: Our purpose was to establish reference values for six monoclonal antibody–based serum immunoassays applied in patients with carcinoma of the female genital tract. STUDY DESIGN: Sera from 938 healthy women (median age 48.0 years, range 37 to 76 years) were assayed for levels of CA 125, CA 15.3 (two methods), CA M29, CA M26, and mucin-like cancer antigen (MCA). RESULTS: Reference values, defined as those including 95% of healthy controls, were in women with unknown menopausal status as follows: CA 125, 37 U/ml; CA 15.3 (Centocor), 33 U/ml; CA 15.3 (Boehringer Mannheim), 28 U/ml; CA M26, 83 U/ml; CA M29, 13 U/ml; and MCA, 19 U/ml. Postmenopausal values were significantly lower for CA 125 and CA M26 and significantly higher for CA M29 and CA 15.3 (both methods). MCA serum levels were age independent. CONCLUSION: Reference values found were not in accord with those generally applied in gynecologic oncology. In addition, serum levels were influenced significantly by menopausal status (except with MCA). (AM J OBSTET GYNECOL 1996;174:107-14.)

Background. Endothelin is involved in the control of cardiovascular and renal functions and acts as a neuromodulator. Methods. In a prospective study among 15 male patients who underwent coronary artery bypass grafting, we investigated the release pattern and possible stimuli of circulating endothelin. Results. We detected a steep increase in endothelin concentrations after the onset of cardiopulmonary bypass (CPB), and a second minor increase during CPB. The steep increase in endothelin concentrations correlated with the change in arterial pressures at the onset of CPB (r = 0.57; p < 0.03). The slow increase in endothelin concentrations during CPB, however, correlated with mean endotoxin levels during and after CPB (r = 0.60; p < 0.02). Conclusions. The change in arterial pressure at the onset of CPB seems to induce a steep and fast increase in circulating endothelin level, which is probably mediated through the baroreceptors. The slow increase in endothelin level during CPB is associated with increased circulating endotoxin concentration. It may be that either endothelin-mediated vasoconstriction induces endotoxin transmigration from the intestine or endotoxin stimulates endothelin secretion from endothelial cells.

1. Uncomplicated insulin-dependent diabetes mellitus is associated with generalized vasodilatation. This vasodilatation is believed to contribute to the development of microvascular complications. The endothelium plays an important role in the regulation of vascular tone. 2. To investigate the role of endothelial mediators, we measured plasma endothelin levels and studied the vascular effects of intravenous l-arginine (the precursor of NO) in 10 male type 1 diabetic patients and 10 non-diabetic subjects. 3. The baseline plasma endothelin level was significantly lower in the diabetic patients [mean 1.7 (SD 0.5) versus 2.1 (0.4) pmol/l; P < 0.05] than in the control subjects. 4. During l-arginine infusion, plasma cyclic GMP (the second messenger for NO) increased in the control subjects [from 5.1 (2.9) to 6.9 (2.9) nmol/l; P < 0.05 versus saline] and in the diabetic patients [from 4.6 (1.8) to 5.7 (2.2) nmol/l; P = 0.09]. l-Citrulline (a by-product of NO synthesis from l-arginine) increased in both groups. The responses to l-arginine were not significantly different between the control subjects and the diabetic patients. The plasma atrial natriuretic peptide level did not change in either group during infusion of l-arginine or of an equal volume of isotonic saline. 5. Blood pressure decreased slightly during l-arginine administration in both groups. In control subjects, the extracellular fluid volume in the lower leg increased during l-arginine infusion as compared with saline; in the diabetic patients both l-arginine and saline increased the extracellular fluid volume. 6. In conclusion, the vascular effects of l-arginine are similar in men with uncomplicated insulin-dependent diabetes mellitus and in control subjects. Whether these effects are due to an increase in NO synthesis remains unclear. The increase in extracellular fluid volume during saline infusion in diabetic patients suggests an increase in microvascular permeability in

A microtiter enzyme-linked immunosorbent assay using recombinant derived antigens was compared with the Western blot (Dupont) in the confirmation of the presence of antibodies against the human immunodeficiency virus type 1 (HIV-1). Of 104 sera (104 individuals) that were negative by a screening ELISA, 91 were also negative by both confirmation assays. In three sera only the microtiter assay was found to be indeterminate, and in nine other sera only Western blot. The only microtiter assay positive serum was from a male patient at risk for infection with HIV. 279 sera from 83 patients were found positive by screening. Of these, 223 sera were positive in both confirmation assays, and no serum was negative. Only one serum was indeterminate by the microtiter ELISA in contrast to 55 sera, including followup samples from 25 patients, most of whom had AIDS, by Western blot (Dupont criteria). However, the number of Western blot indeterminate sera decreased substantially applying less stringent criteria for interpretation. In conclusion, the microtiter ELISA performed well as a confirmation test for the presence of antibodies against HIV-1. In addition, the results demonstrate that in the microtiter assay the envelope peptide kp41 is highly discriminative in detecting anti-HIV-1 negative and anti-HIV-1 positive sera.

In recent years an increasing amount of evidence supports the concept that preeclampsia is an endothelial disease. The purpose of our study was to evaluate the extent to which endothelial cell dysfunction is involved in pathophysiology of preeclampsia.We studied the urinary excretion of thromboxane B2 and 6-keto-prostaglandin F1 alpha and the venous plasma endothelin levels in 23 preeclamptic patients and in control subjects. In six of these patients and in six controls arterial plasma endothelin levels were also measured. In addition, plasma levels of calcitonin gene-related peptide and plasma fibronectin levels were measured. Results were analyzed by Wilcoxon's rank-sum test or signed-rank test.In preeclampsia the urinary thromboxane B2/6-keto-prostaglandin F1 alpha ratio (p0.001), venous plasma endothelin levels (p0.001), and plasma fibronectin levels (p0.001) were significantly elevated compared with normotensive pregnancy. Arterial plasma endothelin levels were significantly higher than venous plasma endothelin levels in normotensive and hypertensive patients (p0.05). Calcitonin gene-related peptide levels showed a wide range in normotensive pregnancy and in preeclampsia, but the difference was not significant.These results confirm the extensive involvement of the endothelium in the pathophysiology of preeclampsia. Preeclamptic vasoconstriction seems to be mediated by an increase in the vasoconstrictor autocoids thromboxane A2 and endothelin. Production of prostacyclin by the vessel wall and endovascular trophoblast might be just a pivotal escape mechanism of the uteroplacental circulation. Calcitonin gene-related peptide appears not to be involved in the pathophysiology of preeclampsia.

To determine whether a sex-associated difference in endothelin levels exists and to assess whether this difference is mediated by sex hormones.Initial survey in healthy persons, followed by a nonrandomized intervention.A university teaching hospital.Twenty-three healthy men, 29 healthy women, 20 pregnant women, and 12 male-to-female and 13 female-to-male transsexual patients.Plasma levels of endothelin were first assessed in healthy men, healthy women, and pregnant women; subsequently, endothelin levels were determined in male-to-female and female-to-male transsexual patients both before and during cross-gender hormone treatment. This treatment involves administration of testosterone esters to women and of ethynylestradiol and cyproterone acetate to men.Endothelin levels were higher in men than in women (5.9 +/- 1.2 compared with 4.17 +/- 0.67 pg/mL; P0.01). Endothelin levels were lower in pregnant women than in age- and sex-matched nonpregnant controls (2.19 +/- 0.73 compared with 4.17 +/- 0.67 pg/mL; P0.01). In 12 male-to-female transsexuals treated with estradiol and the progestational agent cyproterone acetate, endothelin levels decreased from 8.1 +/- 3.0 to 5.1 +/- 2.0 pg/mL (P0.01). In 13 female-to-male transsexuals treated with testosterone, endothelin levels increased from 6.2 +/- 1.1 to 7.8 +/- 1.2 pg/mL (P0.01).Sex hormones may modulate plasma endothelin levels, with male hormones raising levels and female hormones lowering them. This finding may be important in explaining sex-associated differences in susceptibility to atherosclerotic cardiovascular disease.

An immune response within the central nervous system (CNS) is reflected by an increase of immunoglobulin concentrations in the cerebrospinal fluid. IgM is considered the first class of immunoglobulins to be increased after infection. Calculation of the IgM and IgG-index reflects immunological activity within the CNS. Insight in this activity is essential in the diagnosis of several neurological diseases, especially neurosyphilis. A new fast solid phase, double sandwich enzyme-immuno assay for the measurement of IgM in the CSF is described and applied in 71 neurovenereological patients.

The isolated and purified polypeptide chains of calf lens α-crystallin were reaggregated in 11 different mixing ratios. The aggregates were studied by immunoelectrophoresis, and in the analytical ultracentrifuge. The deaggregating agent, 6 M urea, was removed by dialysis for 16, 40 and 100 h. In order to avoid the possible loss of low molecular weight material during dialysis, removal of the 6 M urea was also performed on a Sephadex G-10 column. All reaggregates showed the same sedimentation coefficient and their electrophoretic mobilities reflected only in part the mixing ratio of the polypeptide chains. The aggregates with a high quantity of acidic chains did not, or only partially, incorporate the basic chains.

Extracts from epithelial cells, seven successive concentric layers of the cortex, and the nucleus of calf lenses were submitted to gel chromatography on agarose Bio-Gel A-5m. In the mentioned order, corresponding to increasing age of the relevant lens cells, the content of α-crystallin in these extracts decreased significantly whereas the content of a very high-molecular weight fraction, resembling an aggregate composed of the polypeptide chains of α- and β-crystallin, increased markedly. The sedimentation coefficients of the α-crystallin preparations at concentrations of 0·8 mg/ml ranged from 17 to 21 S and passed through a minimum. Gel electrophoresis in polyacrylamide containing 6 m urea revealed significant changes in polypeptide composition. Nevertheless, no clear relation between sedimentation coefficients and polypeptide composition was found. Evidence has been obtained for the interaction of α-crystallin with constituents of β-crystallin which may lead to the formation of very high-molecular, finally insoluble aggregates.

1. 1. Lenses of bovine embryos and fetuses, ranging in age from 6 weeks to 9 months, were investigated. 2. 2. Weight, shape, protein content and protein composition as well as leucine aminopeptidase activity were determined as a function of age. 3. 3. Alpha-crystallin turned out to be the most prominent embryonic crystallin. 4. 4. The complexity of the protein composition of the lens increased in maturity up to birth. 5. 5. The polypeptide composition of the α-crystallins was analyzed by means of isoelectric focusing in polyacrylamide gels. Two polypeptides, specifiic for the embryonic lenses, were found. 6. 6. One of these specific polypeptides appeared to be sensitive to u.v. light.

Lenses of 3-month-old calves were divided into nine concentric layers. The protein extracts of these fractions were submitted to gel chromatography on agarose A-5 m. From the epithelium to the nucleus, the content of α-crystallin decreased significantly, whereas the content of a high molecular weight fraction, resembling α-crystallin in u.v. spectrum as well as polypeptide composition, increased markedly. An analogous phenomenon was observed in the group of the β-crystallins. The proportion of the lower molecular weight fraction decreased and that of the higher molecular weight fraction increased from epithelium to nucleus, i.e. according to the age of the fibre cells. The highest content of γ-crystallin was found in the nucleus. The activity of lens leucine aminopeptidase was found to be highest in the outer cortex fractions, i.e. in the synthetically active area of the lens.

The value of serial determination of the cerebrospinal fluid (CSF) β-2-microglobulin ( β 2 m) level for early detection of acute lymphoblastic leukemia (ALL) in the central nervous system (CNS) has been prospectively studied in 30 children. β 2 m was determined by micro-ELISA assay. Results demonstrated a sensitivity of 40% (95% confidence interval, 5.3–85.3%) and a predictive value of 8% (95% confidence interval, 1.0–26.0%). In post-irradiation syndrome, as well as in viral infection with cytopathological changes of white cells in CSF, β 2 m values proved to be significantly higher than in incipient CNS relapse, and these conditions, or an unknown cause, are more often associated with β 2 m elevation than CNS relapse. No relation was found between CSF white cell counts and β 2 m levels. It is concluded that β 2 m is not an appropriate test for early detection of CNS involvement in children with acute lymphoblastic leukemia.

Antiserum against the αB chain of bovine α-crystallin has been prepared. Several antigens of the bovine lens are compared using this antiserum.

α-Crystallin was isolated from extracts of the epithelium, from seven successive concentric layers of the cortex and the nucleus of calf lenses. In this order the α-crystallin content of the extracts decreased strongly whereas a very high-molecular component with α-crystallin like subunit composition became more and more prominent. Polyacrylamide gel electrophoresis in 6 M urea revealed significant changes in polypeptide composition. In the epithelial cells only trace amounts of the α A 1 and no α B 1 polypeptides were present. The latter ones appeared at a relative late stage of fiber cell development. The sedimentation coefficients of the α-crystallin preparations obtained from the successive lens layers were also determined. Going from epithelium to nucleus the values obtained ranged from 17 to 21 S and passed through a minimum. A clear cut relationship between the polypeptide chain composition and sedimentation coefficient could not be found.