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Changes in the anthropometric status of rural African under-fives during a decade of primary health care.

Protein energy malnutrition (PEM) is a major cause of hospital admissions and death in most impoverished Third World countries. In the Gelukspan Health Ward, in rural Western Transvaal, a primary health care (PHC) programme, based on the GOBI-FFF strategy, commenced in 1980. Community health surveys on African under-five children were regularly undertaken. In 1990, to assess improvements achieved, anthropometric and other data were obtained on a representative series of 926 children. Harvard standards were used prior to 1984, and American NCHS reference standards thereafter. Low weight-for-age fell from 28 to 19%, low height for age from 33 to 17%, and low weight-for-height from 5 to 1%. The percentage with low arm circumference was unaffected, 3 and 4%, respectively. The improvements described are believed to be due in part to the PHC programme. Greater definition of the causes and extent of improvements are needed to optimize the State's present desire to intensify PHC, especially in rural areas.

Anthropometry↗

A standards-based clinical information system for HIV/AIDS.

OBJECTIVE: To create a clinical data repository to interface the Veteran's Administration (VA) Decentralized Hospital Computer Program (DHCP) and a departmental clinical information system for the management of HIV patients. This system supports record-keeping, decision-making, reporting, and analysis. The database development was designed to overcome two impediments to successful implementations of clinical databases: (i) lack of a standard reference data model, and; (ii) lack of a universal standard for medical concept representation. BACKGROUND: Health Level Seven (HL7) is a standard protocol that specifies the implementation of interfaces between two computer applications (sender and receiver) from different vendors or sources of electronic data exchange in the health care environment. This eliminates or substantially reduces the custom interface programming and program maintenance that would otherwise be required. HL7 defines the data to be exchanged, the timing of the interchange, and the communication of errors to the application. The formats are generic in nature and must be configured to meet the needs of the two applications involved. The standard conceptually operates at the seventh level of the ISO model for Open Systems Interconnection (OSI). The OSI simply defines the data elements that are exchanged as abstract messages, and does not prescribe the exact bit stream of the messages that flow over the network. Lower level network software developed according to the OSI model may be used to encode and decode the actual bit stream. The OSI protocols are not universally implemented and, therefore, a set of encoding rules for defining the exact representation of a message must be specified. The VA has created an HL7 module to assist DHCP applications in exchanging health care information with other applications using the HL7 protocol. The DHCP HL7 module consists of a set of utility routines and files that provide a generic interface to the HL7 protocol for all DHCP applications. SETTING: The VA's DHCP core modules are in standard use at 169 hospitals, and the role of the VA system in health care delivery has been discussed elsewhere. This development was performed at the Miami VA Medical Center Special Immunology Unit, where a database was created for an HIV patient registry in 1987. Over 2,300 patient have been entered into a database that supports a problem-oriented summary of the patient's clinical record. The interface to the VA DHCP was designed and implemented to capture information from the patient treatment file, pharmacy, laboratory, radiology, and other modules. RESULTS: We obtained a suite of programs for implementing the HL7 encoding rules from Columbia-Presbyterian Medical Center in New York, written in ANSI C. This toolkit isolates our application programs from the details of the HL7 encoding rules, and allows them to deal with abstract messages and the programming level. While HL7 has become a standard for healthcare message exchange, SQL (Structured Query Language) is the standard for database definition, data manipulation, and query. The target database (Stitt F.W. The Problem-Oriented Medical Synopsis: a patient-centered clinical information system. Proc 17 SCAMC. 1993:88-93) provides clinical workstation functionality. Medical concepts are encoded using a preferred terminology derived from over 15 sources that include the Unified Medical Language System and SNOMed International ( Stitt F.W. The Problem-Oriented Medical Synopsis: coding, indexing, and classification sub-model. Proc 18 SCAMC, 1994: in press). The databases were modeled using the Information Engineering CASE tools, and were written using relational database utilities, including embedded SQL in C (ESQL/C). We linked ESQL/C programs to the HL7 toolkit to allow data to be inserted, deleted, or updated, under transaction control. A graphical format will be used to display the entity-rel

Acquired Immunodeficiency Syndrome↗

Is quantitative assessment of insulin-antibodies and autoantibodies feasible?

Nine selected sera were studied using radioimmunoassay and enzyme linked immunosorbent assay; eight contained insulin antibodies and were from Type 1 (insulin-dependent) diabetic patients, one of whom had antibody-mediated insulin resistance, and one contained insulin-autoantibodies and was from an asymptomatic blood donor. Sera were assayed in serial dilution to assess their suitability for use as reference standards. Dilution curves were non-parallel in radioimmunoassay but were parallel in immunosorbent assay. In all sera, insulin antibodies were readily detected in both assays whereas the low avidity insulin autoantibodies were only detected by immunosorbent assay and not at all by radioimmunoassay, suggesting that the assays respond differently to antibodies of different avidity. Avidity was estimated in liquid phase from the dissociation rate of preformed complexes of antibody and 125-iodinated insulin. When high avidity antibodies are used as a reference in radioimmunoassay, lower avidity antibodies are underestimated and vice versa. In contrast, in immunosorbent assay, any serum could be used as a reference regardless of avidity; furthermore competition experiments comparing the highest avidity insulin antibodies, from the insulin-resistant patient, with the insulin autoantibodies from the asymptomatic blood donor yielded near-superimposable curves. We conclude that radioimmunoassay is selective for high avidity antibodies whereas enzyme linked immunosorbent assay is not; computer modelling of the two assays supports this conclusion. In practice immunosorbent assay can be standardized using a reference serum, whereas experimental findings and mathematical considerations preclude the use of a standard serum in radioimmunoassay.

Animals↗

Precise measurement of vertebral bone density using computed tomography without the use of an external reference phantom.

Bone density measurement by quantitative computed tomography (QCT) commonly uses an external reference phantom to decrease scan-to-scan and scanner-to-scanner variability. However, the peripheral location of these phantoms and other phantom variables is also responsible for a measurable degradation in accuracy and precision. Due to non-uniform artifacts such as beam hardening, scatter, and volume averaging, the ideal reference phantom should be as close to the target tissue as possible. This investigation developed and tested a computer program that uses paraspinal muscle and fat tissue as internal reference standards in an effort to eliminate the need for an external phantom. Because of their proximity, these internal reference tissues can be assumed to reflect more accurately the local changes in the x-ray spectra and scatter distribution at the target tissue. A user interactive computerized histogram plotting technique enabled the derivation of reproducible CT numbers for muscle, fat, and trabecular bone. Preliminary results indicate that the use of internal reference tissues with the histogram technique may improve reproducibility of scan-to-scan measurements as well as inter-scanner precision. Reproducibility studies on 165 images with intentional region-of-interest (ROI) mispositioning of 1.5, 2.5, or 3.5 mm yielded a precision of better than 1% for normals and 1% to 2% for osteoporotic patients--a twofold improvement over the precision from similar tests using the standard technique with an external reference phantom. Such improvements in precision are essential for QCT to be clinically useful as a noninvasive modality for measurement of the very small annual changes in bone mineral density.

Bone Density↗

The varying distribution of intra- and inter-vertebral height ratios determines the prevalence of vertebral fractures.

Credible inferences regarding the burden of vertebral fractures (VFs) cannot be made without a globally accepted quantitative definition of 'fracture'. Currently, differences in anterior, middle, or posterior vertebral heights (VHs) within a vertebra, or between adjacent vertebrae, are used to define 'fracture'. However, VH differences are essential for the construction of thoracolumbar curves, evolutionary adaptations that provide stability in bipedal stance and gait. As there is no reference standard to distinguish anatomical variation from fracture, approaches to defining a VF use a reference range of VH ratios derived in premenopausal women or derived by trimming, a method that iteratively removes the tails of a distribution of VH ratios to produce a normal distribution. From this, reference ranges of VH ratio means and standard deviations (SDs) are obtained and a nominal deviation of 15% or more, or 3 SD or more is regarded as a 'fracture'. We measured VHs by quantitative vertebral morphometry (QVM) and bone mineral density (BMD) by dual energy X-ray absorptiometry in 697 Lebanese women (age 20-89 years) to compare the prevalence of VF ascertained by published methods and a new method that uses the premenopausal range (without trimming) and requires two VH abnormalities. VF prevalence using published methods reached 60% to 70% in pre- and post-menopausal women, and in women with normal or high BMD because VH ratios were not normally distributed and cut-offs used to define VF fracture fell within the observed distribution of the data. The new method resulted in a VF prevalence of 3.3% in younger and 14% in older women, 7% (high), 10% (middle), and 20% (low) BMD tertiles consistent with the notion that the method detected VF due to bone fragility. We suggest that using a fixed trimming method to define reference range and cut-offs or applying fixed cut-offs to identify VFs in populations, where these ratios are not normally distributed, may result in the capture of anatomical variation, not structural failure. Thus, group differences in the VF prevalence may reflect differences in methodology, not bone fragility. Improved criteria to define VF are needed before credible inferences can be made regarding the burden of VFs in women and men, and between sexes, races, countries, decades, and placebo arms of clinical trials.

Absorptiometry, Photon↗

Simplified method for purification of colostrum to obtain secretory component of immunoglobulin A, using secretory component as a reference protein in tracheal aspirate fluid.

Many studies employ bronchoalveolar lavage fluid for assessment of biologically active substances secreted from the lung. However, investigators continue to search for a useful reference standard to correct for the inevitable but variable degree of dilution of this fluid. The glycoprotein, soluble secretory component of IgA, may serve as a valid reference protein. We report a simplified method for the purification of secretory component from colostrum. Soluble secretory component was isolated from human colostrum using serial centrifugation, size-exclusion fractionation and ion-exchange chromatography. Secretory component rich fractions were assayed by enzyme immunoassay. They were also evaluated for total amino acid content and distribution and sequence determination with satisfactory agreement with published results. We then demonstrated that soluble secretory component concentration in tracheal aspirate fluid did not correlate with either albumin or with total protein measured in the same samples. Therefore, we conclude that the secretory component of IgA serves as a useful reference marker because its use may avoid errors resulting from leakage of plasma proteins into epithelial lining fluid. Advantages of this method for establishing a standard for secretory component include ready availability of soluble secretory component, simplicity of the method and relative rapidity of the techniques.

Chemical Fractionation↗

Assessing the use of known mutagens to calibrate the Salmonella typhimurium mutagenicity assay: II. With exogenous activation.

In order to determine the usefulness of selected chemicals as potential reference materials for calibrating the Salmonella assay, two laboratories tested a series of Salmonella mutagens that require exogenous activation. When the variance for individual substances within a bioassay is sufficiently low and the rankings of those substances are of acceptable consistency, they can later be evaluated for use as standard control compounds, as audit materials, and as standard reference materials for comparative bioassay efforts. The purpose of this project, therefore, was to evaluate the variability in the mutagenic response of potential reference chemicals that require exogenous metabolic activation in the standard plate-incorporation Salmonella mutagenicity assay, and to develop ranking criteria for mutagenic activity based on these data. Ten indirect-acting mutagens were tested in two laboratories using Salmonella typhimurium TA100 and an Aroclor-induced rat liver S9. Each laboratory conducted four definitive testing rounds. A different batch of S9 was utilized for every two rounds. Of the 10 chemicals tested only 2-anthramine had a mean slope value greater than 1000 revertants/micrograms. Three chemicals had slope values between 1000 and 100; and five chemicals had slope values between 100 and 10. The remaining compound, 9,10-dimethyl-1,2-benz[a]anthracene, could not be placed into a single category because it had slope values on either side of 100 revertants per mg. Coefficients of variance were low (i.e., below 25% in most cases). The low variability achieved in this study may be accounted for by two parameters of the study. First, based on Claxton et al. (1991a) and the S9 optimization for three compounds, the amount of S9 was calibrated to a set amount of protein per plate (1.1 mg/plate). Secondly, the 10 test doses were placed in the initial, linear, nontoxic portion of the dose-response curves. The use of ten closely spaced, nontoxic doses allowed for a more accurate estimate of the slope.

Animals↗

Trace level determination of perchlorate in water matrices and human urine using ESI-FAIMS-MS.

High-field asymmetric waveform ion mobility spectrometry (FAIMS) separates perchlorate from interfering isobaric ions of bisulfate and dihydrogenphosphate in the gas-phase. The use of a new FAIMS prototype and waveform generator, along with the use of a mixed carrier gas, in this electrospray ionisation (ESI)-FAIMS-mass spectrometry (MS) study gave a detection limit for perchlorate in a relatively "clean" matrix of tap water of 0.050 ppb. Flow injection analysis (FIA) of dilutions of fortified waste water, a fortified river water certified reference material (CRM; SLRS-4, National Research Council of Canada), and a fortified human urine Standard Reference Material (SRM; 2381, National Institute of Standards and Technology) gave detection limits of 0.37 ppb, 0.50 ppb, and 4.8 ppb, respectively, in the undiluted matrices.

Environmental Monitoring↗

Serum standards for the bioassay of aminoglycosides in cerebrospinal fluid.

Four diluents were compared as reference standards for the assay of gentamicin in cerebrospinal fluid (CSF): human CSF, human serum, distilled water, and 150 mmol NaCl/4.5 mmol CaCl2. Standards prepared in pooled human serum were the best alternative to CSF for the assay of gentamicin and were also useful for the assay of tobramycin, netilmicin, amikacin, and sisomicin. The pH (6.0-9.8) of CSF did not alter the results of the assay.

Aminoglycosides↗

Reference distributions for apolipoproteins AI and B and the apolipoprotein B/AI ratios: a practical and clinically relevant approach in a large cohort.

The two serum apolipoproteins in the highest concentrations, apolipoprotein (apo) AI and apolipoprotein B, and the apolipoprotein B/AI ratio are measured to assess clinical risk for atherosclerotic heart and peripheral vascular diseases. The study is based on a cohort of over 37,000 Caucasian individuals from northern New England measured in one laboratory by immunonephelometry using standardized reference materials. All samples received for protein analyses were accepted provided adequate identifying information was available. Laboratory and demographic information was entered into a single database for subsequent study. Our results show that for males without evidence of inflammation, values of apo AI change little through life. For females, however, values gradually increase until about 60 years of age then fall somewhat thereafter. Among adults, females have higher apo AI values on average, than males. Apo B values change significantly through life, increasing after the end of the second decade to a peak during the sixth decade, then falling thereafter. In the past, concern has been expressed that apo AI is an acute phase reactant (APR), thus complicating cardiovascular risk assessment. The effects of an APR (C-reactive protein >or=10 mg/L) on apo AI, but not on apo B, are measurable for both sexes, most noticeably beyond the age of 60 years in males and females. When values were expressed as age- and gender-specific multiples of the median (MoMs), the resulting distributions fit a log-Gaussian distribution well over a broad range. The size of the relatively homogenous cohort, by a standardized approach, provides a firm basis for comparison to preexisting reference intervals and for establishing a clinically useful and current reference interval for the three main apolipoprotein values.

Age Distribution↗

Performance of a predictive model for streptococcal pharyngitis in children.

CONTEXT: Group A beta-hemolytic streptococcus (GABHS) pharyngitis is a common childhood illness. The clinical diagnosis is difficult to determine and laboratory tests have limitations; hence, the condition is generally overdiagnosed and overtreated. Several clinical pediatric-specific predictive models have been published but none have been prospectively studied. OBJECTIVE: To test the performance of a previously published predictive model for GABHS pharyngitis in children in different clinical settings and during different seasons. DESIGN: Prospective cohort study. SETTINGS: Pediatric emergency department and 2 pediatric outpatient clinics. PATIENTS: Children aged between 1 and 18 years with pharyngitis on initial examination at study sites between April 1, 1999, and March 31, 2000. INTERVENTIONS: Recording of clinical features during initial evaluation using a standardized form and recovery of GABHS from patients' throats using reference standard methods. MAIN OUTCOME MEASURES: Posttest probability for GABHS positive throat culture associated with the model's positive predictors (moderate to severe tonsillar swelling, cervical lymphadenopathy [moderate to severe tenderness and enlargement of cervical lymph nodes], scarletiniform rash, and the absence of coryza) and the models' negative predictors (absence of the above signs and the presence of coryza). RESULTS: Of 587 patients analyzed, 218 (37%) had a positive throat culture for GABHS. Forty-nine percent were boys. Mean +/- SD age was 6.7 +/- 3.9 years. There was no difference between the subsets within the sample. The posttest probability values for a positive throat culture associated with positive and negative predictors of the model were 79% and 12%, respectively. CONCLUSIONS: A pediatric predictive model for GABHS pharyngitis performed better than physicians' subjective estimates for a positive throat culture and was comparable with a rapid antigen detection test. The model performed consistently well in different populations and across seasons. It can be useful if reliable microbiological testing and/or follow-up are not attainable.

Adolescent↗

A comparison of serum electrolyte results by Technicon's Flame III and Flame IV AutoAnalyser system.

A comparison of Technicon's (Technicon, Tarrytown, N. Y., 10591) Flame III and Flame IV electrolyte Auto-Analyzer systems for the determination of Na+, K+, Cl- and CO2 in serum is described. Results from the Flame IV are determined over three periods of time and compared to those of the referee Flame III system. The Optional Digital Printer and Linearizer Modules are not used with the described system. The Flame IV AutoAnalyzer is calibrated with a multi-point calibration technique (aqueous reference standards) rather than the single-point calibration technique (albumin reference material) used by the AutoAnalyzer II. Results for the Flame IV are shown to be comparable with those for the Flame III.

Autoanalysis↗

Sensitivity and specificity of the Swedish interactive threshold algorithm for glaucomatous visual field defects.

PURPOSE: To determine the sensitivity and specificity of two new visual field algorithms in detecting glaucomatous visual field defects: (1) Swedish interactive threshold algorithm (SITA) standard and (2) SITA fast. DESIGN: Prospective observational case series. PARTICIPANTS: Ninety normal subjects and 82 glaucoma patients. TESTING: Central 30 degrees fields were performed with the Humphrey visual field analyzer 30-2 program (Humphrey Systems, Dublin, CA) using full threshold, SITA standard, and SITA fast algorithms on the same day for two or more sessions within a 1-month period. MAIN OUTCOME MEASURES: Sensitivity and specificity in detecting glaucomatous visual field defects with SITA standard and SITA fast using full threshold testing as the reference standard. RESULTS: The sensitivity of SITA standard and SITA fast in detecting glaucomatous defects overall was 98% and 95%, respectively. In the subset of mild glaucomatous field defects (26 patients), sensitivity of SITA standard was 92% versus 85% with SITA fast. Sensitivity was 100% for both algorithms in moderate to severe glaucomatous defects. Specificity for glaucoma defects using SITA standard and SITA fast was 96% for both algorithms. SITA standard reduced test-taking time from full threshold by 52% in normal subjects and 47% in glaucoma patients (P < 0.001). SITA fast reduced test-taking time by 72% in normal subjects and 65% in glaucoma patients (P < 0.001). Mean deviation values were 0.4 dB and 0.8 dB better in SITA standard and SITA fast fields, respectively, in normal subjects (P < 0.001), and 0.7 dB and 1.2 dB in SITA standard and SITA fast fields, respectively, in glaucoma patients (P < 0.001) compared with full threshold values. CONCLUSIONS: The new algorithms for measuring visual fields, SITA standard and SITA fast, have excellent sensitivity and specificity for glaucomatous visual field loss with considerable savings in time.

Adult↗

Immunoglobulin class and subclass concentrations after treatment of childhood leukemia.

With greatly increased survival rates after childhood leukemia during the last 3 decades, the long-term effects of the treatment have become more evident. The disease and its treatment impair the immune system, but the duration of this impairment is unknown. The authors studied the serum concentrations of immunoglobulins and IgG subclasses in 20 Icelandic children cured of leukemia on average 8 years and 3 months after their treatment ended. Although no marked deviations were found in the concentrations of the main immunoglobulin classes IgA, IgM, IgG, and IgE, the IgG subclass levels were below reference values. The patients had on average 0.9 of age standardized reference values of IgG1, 0.5 of IgG2, 0.8 of IgG3, and 0.7 of IgG4. However, none had any autoimmune diseases or a markedly increased tendency for infections. The results indicate that although the immunoglobulin classes regain their normal values within a few years after cessation of treatment, recovery of the IgG subclasses, especially IgG2, is impaired.

Adolescent↗

Prevalence and causes of anemia in the United States, 1976 to 1980.

We estimated the prevalence of anemia in the United States from the results of the Second National Health and Nutrition Examination Survey ( NHANES II, 1976 to 1980). Reference ranges for Hb were first derived from 11,547 subjects in whom laboratory values for serum iron/iron-binding capacity, mean corpuscular volume, and erythrocyte protoporphyrin were all normal (greater than or equal to 16%, greater than or equal to 80 fl, and less than or equal to 75 micrograms/dl red blood cells, respectively). Using these reference standards, the prevalence of anemia (Hb values below the 95% reference range for age and sex) among the 15,093 subjects with complete laboratory results was highest in infants (5.7%), teenage girls (5.9%), young women (5.8%), and elderly men (4.4%). The pattern of laboratory abnormalities in anemic subjects indicated that iron deficiency predominated as a cause in infants and young women in contrast to inflammatory disease in the elderly.

Adolescent↗

Secondary cleft sign as a marker of injury in athletes with groin pain: MR image appearance and interpretation.

PURPOSE: To determine whether the secondary cleft sign demonstrated in the symphysis pubis at magnetic resonance (MR) imaging is a marker of injury in athletes presenting with groin pain. MATERIALS AND METHODS: Ethics review board approval was not required for studies involving retrospective image or case record review; informed consent for review was not required. Eighteen male athletes (mean age, 24 years; age range, 19-32 years) were included for study. All patients underwent radiography and MR imaging (coronal fast spin-echo T1-weighted, transverse fast spin-echo T2-weighted, and coronal turbo short inversion time inversion-recovery [STIR] imaging) of the pelvis. Subsequent image-guided nonionic contrast material injection was followed by a 0.5% bupivacaine hydrochloride (1 mL) and methyprednisolone acetate (20 mg) injection into the central cleft of the symphysis pubis. Comparison was made between imaging findings at symphyseal cleft injection and appearances at preprocedure MR imaging, with specific reference to the presence of a secondary cleft. The sensitivity and specificity of MR imaging in demonstrating the secondary cleft sign were compared with those of the reference standard, imaging at symphyseal cleft injection. MR images from a reference group of 70 asymptomatic athletes who underwent STIR imaging of the pelvis were analyzed for evidence of a secondary cleft. RESULTS: Osteitis pubis was diagnosed in six patients on the basis of radiography and/or MR imaging. A secondary cleft was identified in 12 of 18 patients at MR imaging, was best visualized at coronal STIR imaging, and was confirmed in each patient during contrast material injection into the central physiologic symphyseal cleft. In no patient was a secondary cleft identified at symphyseal cleft injection and not identified at MR imaging (sensitivity and specificity, 100%). In each patient, the side of the secondary cleft corresponded to the side of symptoms that responded to local anesthetic and steroid injection. Four of the six patients with osteitis pubis had evidence of a secondary cleft. In one patient, a secondary cleft was not identified at MR imaging or symphyseal cleft injection, but adductor avulsion was identified at MR imaging. No evidence of a secondary cleft sign at MR imaging was identified in the reference group. CONCLUSION: The secondary cleft sign demonstrated at MR imaging is a marker of groin injury in athletes presenting with groin pain.

Adult↗

Use of inductively coupled plasma-mass spectrometry in boron-10 stable isotope experiments with plants, rats, and humans.

The commercial availability of inductively coupled plasma-mass spectrometry technology (ICP-MS) has presented the opportunity to measure the boron concentrations and isotope ratios in a large number of samples with minimal sample preparation. A typical analytical sequence for fecal samples consists of 25 acid blanks, 1 digestion blank, 5 calibration solutions, 4 standard reference material solutions, 10 samples, and 4 natural abundance bias standards. Boron detection limits (3 x 1 sigma) for acid blanks are 0.11 ppb for 10B, and 0.40 ppb for 11B. Isotope ratios were measured in fecal samples with 20 to 50 ppb boron with < 2% relative standard deviation. Rapid washout and minimal memory effects were observed for a 50 ppb beryllium internal standard, but a 200 ppb boron biological sample had a 1.0 ppb boron memory after a 6-min washout. Boron isotope ratios in geological materials are highly variable; apparently this variability is reflected in plants of a fixed natural abundance value for boron requires that a natural abundance ratio be determined for each sample or related data set. The natural abundance variability also prevents quantitation and calculation of isotope dilution by instrument-supplied software. To measure boron transport in animal systems, 20 micrograms of 10B were fed to a fasted rat. During the 3 days after a 10B oral dose, 95% of the 10B was recovered from the urine and 4% from the feces. Urinary isotope ratios, 11B/10B, changed from a natural abundance of 4.1140 to an enriched value of 0.95077, a 77% change.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Biologically active luteinizing hormone (LH) in plasma: II. Comparison with immunologically active LH levels throughout the human menstrual cycle.

The levels of biologically active luteinizing hormone were determined by an in vitro bioassay method in plasma samples collected daily over a complete menstrual cycle from 12 menstruating women. These cycles were normal according to a number of criteria, including daily plasma levels of oestradiol, 17-hydroxyprogesterone and progesterone. Immunoreactive LH was estimated in the same 12 cycles by a radio-immunoassay (RIA) procedure (HCG-RIA) using an HCG antiserum and iodinated HCG. The 2nd IRP of HMG was selected as standard although significant deviations from parallelism were found with 7 out of the 12 plasma pools studied. The use of the 1st IRP of human pituitary gonadotrophins (FSH and LH (ICSH)) for bioassay (herafter HPG-1st IRP) as standard in this system resulted invariably in invalid assays, due to lack of parallelism. Immunoreactive LH was also measured in 8 of the 12 cycles by a RIA procedure (HLA-RIA) using a human LH antiserum and iodinated human LH of pituitary origin. Results are expressed in terms of the HPG-1st IRP. The plasma levels of biologically and immunologically active LH were qualitatively similar throughout the menstrual cycle. However, the LH levels measured by the bioassay invariably exceeded those estimated by the RIA procedures. The biological to immunological (B/I) ratio over the entire menstrual cycle (312 comparisons) was 5.5 with 95% confidence limits at 5.2 and 5.8 when the HCG-RIA system was employed. Using the HLH-RIA system (208 comparisons), the corresponding ratio was 6.4 (6.0:6.9). When regression lines were calculated using the bioassay results as the independent variable and the RIA results as the dependent variable, the 95% confidence limits of the regression lines did not include the origin. Furthermore,, in keeping with the high B/I ratios, the slopes of the two regression lines and their conficence limits differed markedly from unity. It is concluded that although qualitatively similar profiles were observed between the biological and immunological activities throughtout the menstrual cycle, two aspects require further attention. Firstly, the elevated B/I ratios together with the behaviour of the dose-effect lines obtained with different standards in the various RIA systems suggest that presently available reference standard preparations of pituitary and/or urinary origin are not suitable for the assay of LH in human plasms. Secondly, from the regression analyses of the biological and immunological activities it is infered that the RIA methods detect immunological activity which is not associated with biological activity. If so, the validity of these RIA procedures for specifically measuring low levels of biologically active LH in plasma may be in question.

Adult↗