Effect of timing of relocation of replacement gilts from group pens to individual stalls before breeding on fertility and well-being.

Variation in gilt fertility is associated with increased replacement and reduced longevity. Stress before breeding is hypothesized to be involved in reduced fertility. This study tested the timing of gilt relocation from pens to individual stalls before breeding on fertility and well-being. The experiment was performed in replicates on a commercial research farm. After detection of first estrus, gilts ( = 563) were assigned to treatment for relocation into stalls 3 wk (REL3wk), 2 wk (REL2wk), or 1 wk (REL1wk) before breeding at second estrus. Subsets of gilts from each treatment ( = 60) were selected for assessment of follicles at second estrus. Data included interestrus interval, number of services, conception, farrowing, total born, and wean to service interval. Piglet birth weight was obtained on subsets of litters ( = 42/treatment). Measures of well-being included BW, backfat, BCS, lesions, and lameness from wk 1 after first estrus until wk 16. Gilt BW at wk 5 (158.4 kg) was not affected ( > 0.10) by treatment. Measures of BCS, lameness, and lesions at breeding and throughout gestation did not differ ( > 0.10). Treatment did not affect ( > 0.10) gilts expressing a normal interestrus interval of 18 to 24 d (83.4%) but did influence ( < 0.05) the proportion expressing shorter ( < 0.001) and longer ( < 0.001) intervals. Gilts in REL3wk had a shorter ( < 0.001) interestrus interval (20.7 d) than those in REL2wk and REL1wk (22.6 d). Gilts with shorter intervals ( = 24) had fewer total born while gilts expressing longer cycles ( = 65) had reduced farrowing rates. The number of services (1.9) and number of follicles (19.7) at breeding were not affected ( > 0.10) by relocation. There was no effect of treatment on farrowing rate (85.2%), born alive (12.6), or any litter birth weight measures ( > 0.10). The percentage of sows bred within 7 d after weaning (94.4%) was also not affected by treatment ( > 0.10). These results suggest that the timing of relocation before breeding had no effect on well-being or on the majority of gilts with normal estrous cycles and their subsequent fertility. However, a smaller proportion of the gilts exhibited shorter and longer interestrus intervals in response to relocation 1 or 3 wk before breeding. In cases where gilt fertility may be less than optimal, producers that relocate gilts from pens to stalls before breeding should evaluate interestrus interval as a response criterion.

Hyperglycosylated HCG expression in pregnancy: cellular origin and clinical applications.

Employing a monoclonal antibody (B152) specific for a carbohydrate epitope found on a choriocarcinoma derived hCG, it was discovered that a similar hCG isoform is expressed during early pregnancy. This form differs from later pregnancy hCG in carbohydrate moieties. Profiling of these two hCG isoforms throughout pregnancy utilized two IRMA's: B152-B207 ("hyperglycosylated hCG"-specific assay) and B109-B108 (an IRMA for standard intact hCG isoforms in the WHO hCG reference preparation). The WHO hCG standard was used in both assays. Values were presented as a ratio of hCG isoform concentrations (B152/B109 ratio). In early pregnancy urine concentrations of B152 hCG were significantly higher in normal pregnancy (NP) compared to early pregnancy loss (EPL). Matched serum-urine samples from the first and third trimesters revealed that the B152 hCG form is predominant in both serum and urine in the first trimester compared with the third trimester. The proportion of the B152 hCG (HhCG) form is higher in urine than in matched serum. There was a significant difference in the B152/B109 ratio between days 5 and 20 from time of embryo transfer in normally developing pregnancy versus EPL in the urine of IVF patients. In spontaneous abortion (SA) the level of B109 hCG remained higher in NP compared with SA. However, the B152/B109 ratio declined with gestational age faster in SA than in NP suggesting perhaps a different loss mechanism in SA versus EPL. The cellular origin of the different hCG glycoforms was identified by assay of cell media from cytotrophoblasts (CTBs) and syncytiotrophoblasts (STBs). Isolated CTBs expressed predominantly HhCG. The level of expression was the highest in the first trimester. STBs were the source of the less glycosylated B109 hCG isoform. Analysis of hCG glycoforms during early pregnancy can distinguish pregnancies that will fail from those that will proceed normally. Since the B152 assay does not effectively discriminate between intact HhCG and free beta HhCG (HhCGbeta), a new HhCGbeta assay was developed. This assay recognizes the HhCGbeta which is produced by CTBs. We hypothesize that the measurement of HhCGbeta may have a potential use in screening for Down syndrome and perhaps other pregnancy disorders and certain types of cancer.

Impact of biological and economic variables on optimal parity for replacement in swine breed-to-wean herds.

Voluntary and involuntary culling practices determine the average parity when sows are replaced in a herd. Underlying these practices is the economic effect of replacing a sow at different parities. A dynamic programming model was used to find the optimal parity and net present value in breed-to-wean swine herds. The model included income and costs per parity weighted by the discount rate and sow removal rate. Three scenarios that reflect a wide range of cases were considered: low removal rates per parity with no salvage value (LRNS), high removal rates per parity with no salvage value (HRNS), and high removal rates per parity with a percentage of the sows having a salvage value (HRYS). The optimal parity of replacement for the base biological and economic conditions was 4 and 5 parities in the high and low removal scenarios, respectively. Sensitivity analyses identified the variables influencing the optimal replacement parity. Optimal parity of replacement ranged from 3 to 7 parities in the low replacement scenario, compared with 1 to 5 parities in the high replacement scenarios. Sow replacement cost and salvage value had the greatest impact on optimal parity of replacement followed by revenues per piglet weaned. The discount rate and number of parities per year generally had little influence on optimal parity. For situations with high sow costs, low salvage values, and low revenues per piglet, the optimal parity at removal was as high as 6 to 10 parities, and for situations with low sow cost, high salvage values, and high revenues per piglet, the optimal parity at removal was as low as 1 to 2 parities depending on removal rates. The modified internal rate of return suggested that, for most LRNS and HRYS scenarios considered, investment in a swine breed-to-wean enterprise was favored over other investments involving a similar risk profile. Our results indicate that in US breeding herds, sows are culled on average near the optimal parity of 4. However, the optimization process should be a dynamic one that adapts to changes in replacement rates, salvage value, replacement cost, and revenues per piglet.

Bioeconomic evaluation of sow longevity and profitability.

Sow production indicators, including litter size, litter weight, and the length of time that sows remained in the herd (sow longevity), were used to characterize sow performance and profitability. Sow longevity and production records from 148,568 sows in 32 commercial herds from Central Illinois from January 1995 to May 2001 were analyzed using survival and repeatability models, respectively. The factors studied included sow genetics (32 genetic lines), with eight major lines present in multiple herds, and the combination of herd and year of entry in the herd. The largest difference in longevity between the major genetic lines was approximately one parity. There were differences (P < 0.05) in the instantaneous sow removal rate or hazard from the major lines. These differences constitute evidence that sow longevity could be improved by using replacements from specific genetic lines. The net present value per sow (present value of future cash flows and the present value of the sow) was used to evaluate the effect of sow longevity and production traits on economic returns. Assuming a zero discount rate per parity, genetic lines with longer herd life resulted in greater profit than genetic lines with shorter herd life. This difference was reduced with increasing discount rates and was reversed with high discount rates and low net income per litter. These results suggest that the magnitude of the economic improvement attained through the use of sow genetic lines with longer longevity depends on the economic context under which the evaluation is made.

Trophoblast origin of hCG isoforms: cytotrophoblasts are the primary source of choriocarcinoma-like hCG.

We have previously demonstrated that a hyperglycosylated isoform of chorionic gonadotropin (hCG) (B152 hCG) is detected in the blood and urine in early pregnancy and is subsequently rapidly replaced by the hCG isoform (B109 hCG) characteristic of later pregnancy. In the current study we have extended our work on the origin of these isoforms. We have used a combination of in situ and in vitro approaches. Localization studies in placental tissues showed that monoclonal antibody B109 stained very specifically syncytiotrophoblast (STBs) from first and second trimester tissues. At term, STBs exhibited no B109 staining at all. Immunostaining with B152 antibody, that recognize the hyperglycosylated isoform of hCG, revealed only punctate staining of STBs in most villi of first trimester tissue. Both antibodies B109 and B152 failed to stain cytotrophoblasts (CTBs). To assess the functional relevance of these observations we analyzed conditioned media from purified CTBs using two immunometric assays, one of which (B152-B207*) has primary specificity for the hyperglycosylated, choriocarcinoma-like hCG and the other (B109-B108*) having primary specificity for the later pregnancy hCG isoform. Regardless of gestational age, isolated CTBs secreted predominantly B152 hCG isoform in contrast to placental villi (predominantly STBs), which released primarily the B109 hCG isoform. Isolated CTBs, however, failed to immunostain with both B109 and B152 antibodies. To resolve this contradiction, we cultured CTBs in the presence of brefeldin A, a drug known to block secretion by inhibiting protein translocation from the endoplasmic reticulum to the Golgi vesicles. Brefeldin A treated CTBs stained strongly with B109 and did not stain or stained weakly with B152 antibody. We assume that treatment with brefeldin A impaired glycosylation of beta subunit and consequently inhibited the production of hyperglycosylated form of hCG recognized by B152. In summary, our in vitro experiments indicate that both isoforms of hCG are produced by villus CTBs and that the dominant isoform is the one recognized by antibody B152. STBs produce primarily the less glycosylated B109 hCG isoform. This data suggests that at the beginning of pregnancy villus CTBs are the major source of the B152 hCG isoform. This finding is supported by our clinical data that show that the dominant hCG isoform in the blood and urine of pregnant women in the first 6 weeks of pregnancy is recognized by B152 (). The inversion of the B152/B109 ratio observed after 6-7 weeks of pregnancy can be explained by the reduction of number of villus CTBs and/or by maturation of STBs.

Reliability of serial urine HCG as a biomarker to detect early pregnancy loss.

BACKGROUND: To examine the reliability of HCG as a biomarker for early pregnancy loss, five experienced researchers independently assessed data from 153 menstrual cycles, determining whether each cycle represented 'no conception,' a 'continuing conception' or a 'conception lost.' METHODS: Urine samples were analysed by immunoradiometric assay using a combination of capture antibodies for the intact heterodimer (B109) and for an epitope common to the beta subunit and the beta core fragment (B204). For each cycle, HCG data were presented as graphs of daily assay results. Summary statistics for HCG assays from 46 women who had undergone bilateral tubal ligation represented baseline values. RESULTS: Pairwise agreement among the assessors for any of the three options ranged from 78-89%. At least three experts agreed for 147 cycles (96%), accounting for 28 conception losses and 19 continuing conceptions. The multi-rater kappa was 0.62 for the conception lost category and 0.68 for continuing conceptions, indicating substantial agreement. CONCLUSION: The main sources of disagreement involved deciding whether there was sufficient information for assessment, interpreting cycle parameters such as cycle length or bleeding event, and interpreting a distinct HCG rise pattern that does not exceed the baseline value obtained from the sterilized women.

Differential expression of human chorionic gonadotropin (hCG) glycosylation isoforms in failing and continuing pregnancies: preliminary characterization of the hyperglycosylated hCG epitope.

Human chorionic gonadotropin (hCG) glycoforms change as pregnancy progresses. We have developed an antibody (B152) which can measure a hyperglycosylated early pregnancy isoform of hCG. This putative hyperglycosylated form of hCG arises very early in pregnancies and is rapidly replaced by an isoform that predominates for the remainder of the pregnancy. The profiles of these hCG glycoforms are measured as a ratio of values of two immunometric assays. The profiles of these ratios differ between pregnancies which persist and those which will experience early failure. In this report, daily urine hCG isoform ratios from donor eggs (no exogenous hCG pretreatment), in vitro fertilization pregnancies were profiled and analyzed from the first day following embryo transfer (ET). Significant differences were found between continuing pregnancy and pregnancy loss throughout days 5-20 post-ET. When hCG isoform ratios were analyzed from the first day of detectable hCG, pregnancy loss could be predicted in the case of a single fetus both during the 5- to 10-day time segment (P=0.018) and the 10- to 15-day time segment (P=0.045). When single and multiple fetus pregnancies were analyzed together significance was approached in the 10- to 15-day time period (P=0.058). In a second population of pregnant women who conceived naturally, in whom urine samples were collected at approximately weekly intervals to either term birth or clinical spontaneous abortion, the ratio could discriminate between miscarriages and normal term pregnancies (P=0.043). In later pregnancy, the ratio of hCG isoforms declined more rapidly in miscarriages than in term pregnancy. Antibody B152 was produced using a choriocarcinoma-derived hCG (C5), which was hyperglycosylated at both N- and O-linked sites and was 100% nicked at position beta(47-48). Western blot analyses supported the assay results showing that early pregnancy urine does not contain nicked C5-like hCG. Also, the early pregnancy hCG appeared to be the same size as later pregnancy hCG as judged by SDS gel electrophoresis. A series of Western blot analyses and immunoassays conducted with the samples either non-reduced or reduced showed that B152 is directed to a linear epitope located in the COOH-terminal peptide region of the beta subunit. This indicated that only the O-glycan groups and not the N-linked glycans are part of the antibody epitope.

Choriocarcinoma-like human chorionic gonadotrophin (HCG) and HCG bioactivity during the first trimester of pregnancy.

The objective of this study was to evaluate the distribution of choriocarcinoma-like human chorionic gonadotrophin (HCG) isoforms during first trimester pregnancy and their relationship with in-vitro HCG bioactivity. This was done by means of a retrospective analysis of patients' sera with first trimester normal intrauterine and abnormal (ectopic) pregnancies. Serum samples were obtained from 38 women with an amenorrhoea of <10 weeks. From these, 19 had a normal intrauterine pregnancy (IUP) and 19 an ectopic pregnancy (EP). Total immunoreactive HCG (HCGi), free beta-HCGi and oestradiol were measured by enzyme immunoassays and bioactive HCG by the mouse Leydig cell bioassay. The alterations in HCG isoform content were measured by the combination of two immunometric assays, B152 for choriocarcinoma-like HCG and B109 for intact HCG detection and expressed as the B152/B109 ratio. Choriocarcinoma-like HCG isoforms ratio measured by B152 and B109 assays was significantly higher in the low subgroups of free beta-HCGi and gestational age (P = 0.0111 and 0.0036 respectively). Whereas bioactive to immunoreactive HCG ratios (b/i ratio) were significantly higher when free beta-HCGi concentrations were low (P = 0.0010), no correlation was found between the variation of bioactivity (b/i ratio) and the proportion of choriocarcinoma-like HCG isoforms (B159/B108). It is concluded that in first trimester pregnancies (i) the modulation of HCG in-vitro bioactivity is not related to the variation of choriocarcinoma-like HCG isoforms secretion and (ii) the amount of choriocarcinoma-like HCG isoforms secreted by the early trophoblast is predominant and may be the result of an early developmental regulation of glycosylation enzyme.

The cost-effectiveness of strategies to assess gastroesophageal reflux as an exacerbating factor in asthma.

OBJECTIVES: We sought to evaluate the cost-effectiveness of diagnostic strategies to determine whether or not acid reflux exacerbates asthma, and to identify which asthma response probabilities are most important in a cost-effective workup of this problem. METHODS: We performed a cost-effectiveness analysis, comparing 11 diagnostic strategies to assess the role that acid reflux plays in asthma. Probabilities and costs were derived from the published literature. Average and incremental costs, effectiveness, and cost-effectiveness were calculated for each strategy. Sensitivity analyses were performed. RESULTS: The most cost-effective diagnostic approach is to begin with omeprazole 20 mg/day for 3 months, followed by 24-h pH testing on drug in nonresponders. If 24-h pH testing is positive, increase the omeprazole dose every 3 months until the patient responds or a maximum of 60 mg/day is given. This strategy costs $730 per case correctly diagnosed. When the cost of pH testing exceeds $586 or the cost of omeprazole 20 mg/day is <$53 per month, omeprazole 20 mg/day for 3 months followed by 60 mg/day for the same duration in nonresponders becomes more cost-effective. CONCLUSIONS: Empiric acid reflux suppression, followed by pH testing in nonresponders, is the most cost-effective means of determining whether GERD is aggravating a patient's asthma.

Early pregnancy human chorionic gonadotropin (hCG) isoforms measured by an immunometric assay for choriocarcinoma-like hCG.

Human chorionic gonadotropin (hCG) exhibits molecular heterogeneity in both its protein and carbohydrate moieties. This communication describes changes in hCG isoforms detected directly in clinical samples. These isoforms, quantified in blood or urine specimens, show a progression of change throughout normal pregnancy. Early pregnancy produces a type of hCG that resembles, in terms of immunoreactivity, a major form of hCG excreted in choriocarcinoma. The isoforms predominate for the first 5-6 weeks of gestation and then diminish, being replaced with the hCG isoforms which predominate throughout the remainder of pregnancy. The alteration in hCG isoform content occurs in both blood and urine. The progression of isoforms is best delineated by calculating the change in the ratio of the two forms, as many hCG assays either do not detect or fail to discriminate among these isoforms. An analogous pattern of hCG isoforms was observed in patients with in vitro fertilization pregnancies. hCG isolated from the pituitary displayed binding characteristics similar to those of the hCG derived from normal pregnancy urine. The early pregnancy hCG isoforms appear to have a differential expression in normal pregnancy as opposed to pregnancies which will not carry to term, suggesting that a determination of the relative balance of hCG isoforms may have diagnostic application in predicting pregnancy outcome.

Evaluation of nicked human chorionic gonadotropin content in clinical specimens by a specific immunometric assay.

We report the development and characterization of an IRMA for the direct measurement of nicked human chorionic gonadotropin (hCGn) in blood and urine. hCGn derived from a reference preparation of hCG used as an immunogen elicits monoclonal antibodies (mAbs) with enhanced recognition of human luteinizing hormone epitopes. The most specific assay for pregnancy hCGn is an IRMA composed of one mAb to choriocarcinoma-derived hCGn (C5) and a second mAb developed from immunization with normal-pregnancy hCGn. This assay was used to evaluate hCGn profiles in normal, in vitro fertilization, Down syndrome, and ectopic pregnancies. In all pregnancies, hCGn was usually present in much lower concentrations than the non-nicked hCG isoform. Our results suggest that some form of physical separation from the overwhelming quantities of non-nicked hCG present in clinical specimens will be required before accurate immunochemical estimations of hCGn can be made.

Fluoroscopic examination in the neonatal intensive care unit using an inverted C-arm.

Fluoroscopic studies can be performed in the Neonatal Intensive Care Unit (NICU) using a portable C-arm. The C-arm is inverted so that the infant lays directly on the image intensifier. In all our cases, the images obtained were of good enough quality to allow for prompt diagnosis and management of the patient's condition. Furthermore, measured entrance doses were lower using the inverted C-arm than they were using conventional fluoroscopy.

Alcoholic liver disease: proposed recommendations for the American College of Gastroenterology.

OBJECTIVE: The objective of this work was to develop practice guidelines for the management of alcoholic liver disease. METHOD: A computerized search using the Medline Data Base from 1966-July 1997 was performed with the search headings; alcohol, alcoholic hepatitis, alcoholic liver disease, liver transplant, diagnosis, epidemiology, human, and English only. All randomized controlled trials, case-control studies, and meta-analyses were read in depth. A manual search was also done using references from each retrieved report, review articles, editorials, postgraduate course syllabi, and textbooks. In the subsequent review, evidence was evaluated using a hierarchical scale with randomized, controlled trials given the most importance. Abstracts presented at national meetings were included only when unique data were obtained from those studies.

The expression of the urinary forms of human luteinizing hormone beta fragment in various populations as assessed by a specific immunoradiometric assay.

Human gonadotrophins undergo metabolic transformations which result in the presence of several smaller, structurally and immunologically related forms of gonadotrophins in the urine. For luteinizing hormone (LH), a beta core fragment (LHbeta cf) has been isolated from the pituitary and characterized. The corresponding urinary fragment is inferred from mass spectral and immunochemical analysis of chromatographically separated urinary forms. Physicochemical characteristics, primarily mass spectral and chromatographic, indicate that the pituitary and urinary forms of LHbeta cf have a different structure, probably in the carbohydrate moieties. This communication characterizes the expression of LHbeta cf in the urine of both reproductive and post-reproductive age women and in men, employing assays highly specific for the pituitary form of the fragment. It was found that LHbeta cf is the predominant LH associated molecular form in the urine during peri-ovulatory period, peaking 1-3 days later than intact LH and reaching a concentration of approximately 600 fmol/mg creatinine, 7-fold higher than either LH or LH free beta subunit. Corresponding concentrations of human chorionic gonadotrophin (HCG) beta cf were <1% that of LHbeta cf. LHbeta cf cross-reaction with some LH or LHbeta monoclonal antibodies may well interfere with the accurate estimation of the day of the LH surge when urinary tests are utilized.

Measuring human chorionic gonadotropin in the absence of implantation with use of highly sensitive urinary assays for intact beta-core and free beta epitopes.

OBJECTIVE: To determine if human chorionic gonadotropin (hCG) can be absorbed from the uterine cavity in the absence of an embryo. DESIGN: Prospective study. SETTING: University-based assisted reproduction program. PATIENT(S): Eight functionally agonadal patients (age range, 33-46 years) who were taking hormone replacement therapy so that they could receive donated oocytes. INTERVENTION(S): Intrauterine instillation of 50 microL of hCG (10,000 IU) during a mock cycle before an attempt at oocyte donation. MAIN OUTCOME MEASURE(S): Spot urine measurements of different hCG epitopes (intact beta, beta-core, and free beta) at timed intervals (12, 20, 44, and 68 hours after instillation). RESULT(S): All hCG epitopes were detected in the urine at the first sampling interval, and levels decreased in subsequent sampling intervals. Measurement of the serum hCG level confirmed that systemic absorption had occurred and that the urine measurements were not a result of specimen contamination through the cervix. CONCLUSION(S): hCG may be systemically absorbed into the blood through the uterine cavity, even in the absence of implantation, and its metabolites may be measured with use of highly sensitive urinary assays.

Detecting pre-ovulatory luteinizing hormone surges in urine.

The study objectives were to determine (i) if pre-ovulatory luteinizing hormone (LH) surges, undetected in urine by two immunoradiometric assays (IRMA), were detectable by an ultrasensitive immunofluorometric assay (IFMA) and (ii) the influence of creatinine adjustment on the detection and timing of the urinary LH surges. Daily urine specimens were contributed by healthy 25-36 year old volunteers during 14 ovulatory menstrual cycles for an epidemiological study conducted in 1983-1985. Specimens were selected as having been previously assayed by two IRMA without consistently detecting LH surges. These urine specimens were remeasured using an IFMA and adjusted for creatinine concentration. IFMA measurements revealed unambiguous LH surges in all cycles. Adjusting IRMA urinary LH values for creatinine concentrations revealed previously undetected LH surges in four of eight cycles. Creatinine adjustment also altered the timing of IRMA and IFMA LH surges by 1-5 days. These results demonstrate an IFMA that detects pre-ovulatory LH surges in unpreserved, frozen urine from cycles where such surges were previously undetectable. Further, creatinine adjustment can markedly affect detection and timing of the onset and peak of the urinary LH surge. While our analysis suggests that this adjustment improves the validity of the LH measure, this requires further investigation.

Differential urinary gonadotrophin profiles in early pregnancy and early pregnancy loss.

Early pregnancy loss (EPL), detected by patterns of human chorionic gonadotrophin (hCG) in urine, is the biomarker employed in investigations of the impact of personal, workplace or environmental reproductive toxins on human fertility. An issue central to these studies is what, in terms of urinary hCG expression, constitutes an EPL. This report describes the urinary molecular forms of hCG expressed in menstrual cycles in which a normal pregnancy was conceived, or an EPL occurred, or no apparent conception occurred. Qualitative and significant quantitative differences in the expression of hCG-associated analytes were found between normal pregnancy cycles and EPL cycles. Discriminant analysis calculation based on mole fractions of the different hCG-associated molecules afforded 91 per cent and 80 per cent correct classification of clinical pregnancy cycles and EPL cycles, respectively. Although hCG-associated molecules unique to either EPL or normal pregnancy were not found, what is thought to be an early form of hCG is expressed both at a high frequency and at a significantly higher concentration in early normal pregnancy when compared with EPL. The relative absence of this molecule very early in pregnancy may signal a pregnancy loss.

Spinal extramedullary hematopoiesis secondary to hepatocellular carcinoma. Case report and literature review.

A 38-year-old man, with hepatitis C and a history of intravenous drug use had hepatocellular carcinoma. Severe low back pain was the result not of metastases but of extramedullary hematopoiesis. This was not confirmed by biopsy but seemed unequivocal on magnetic resonance imaging.