Ultrasonographic diagnosis of early unruptured tubal pregnancy in a community hospital.

OBJECTIVE: To investigate ultrasonographic characteristics and diagnostic approaches of unruptured tubal pregnancy (UTP). METHODS: One hundred forty-six cases of early tubal pregnancy, which were confirmed surgically, were analyzed. RESULTS: Among 146 cases of UTP, 130 were diagnosed by ultrasonography. Furthermore, 16 cases of UTP were missed by ultrasonography. A total of 130 patients had a mass detected by ultrasonography, among whom the mass and ipsilateral ovary were distinguishable in 102. The embryo sac type was found in 40.00% (52/130), while the heterogeneous mass type was found in 60.00% (78/130) of patients. The maximum length and diameter of the ectopic pregnancy mass ranged from 10 to 68 mm (mean size, 26.42 ± 11.39 mm). No blood flow was observed in or around the mass in 67.69% (88/130) of patients. Lateral tubal thickening of the mass was found in 47.69% (62/130) of patients. Endometrial thickness was ≤10 mm in 67.80% (99/146) of patients. The intima was mostly hyperechoic in 72.60% (106/146) of patients. CONCLUSION: A paraovarian mass, thickened fallopian tube, uterine endometrium with a thickness of ≤10 mm, and high echo are important indicators of early UTP. Ultrasound examinations should focus on these indicators to help improve detection of early tubal pregnancy.

Insights into the hyperglycosylation of human chorionic gonadotropin revealed by glycomics analysis.

Human chorionic gonadotropin (hCG) is a glycoprotein hormone that is essential for the maintenance of pregnancy. Glycosylation of hCG is known to be essential for its biological activity. "Hyperglycosylated" variants secreted during early pregnancy have been proposed to be involved in initial implantation of the embryo and as a potential diagnostic marker for gestational diseases. However, what constitutes "hyperglycosylation" is not yet fully understood. In this study, we perform comparative N-glycomic analysis of hCG expressed in the same individuals during early and late pregnancy to help provide new insights into hCG function, reveal new targets for diagnostics and clarify the identity of hyperglycosylated hCG. hCG was isolated in urine collected from women at 7 weeks and 20 weeks' gestation. hCG was also isolated in urine from women diagnosed with gestational trophoblastic disease (GTD). We used glycomics methodologies including matrix assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometry (MS) and MS/MS methods to characterise the N-glycans associated with hCG purified from the individual samples. The structures identified on the early pregnancy (EP-hCG) and late pregnancy (LP-hCG) samples corresponded to mono-, bi-, tri-, and tetra-antennary N-glycans. A novel finding was the presence of substantial amounts of bisected type N-glycans in pregnancy hCG samples, which were present at much lower levels in GTD samples. A second novel observation was the presence of abundant LewisX antigens on the bisected N-glycans. GTD-hCG had fewer glycoforms which constituted a subset of those found in normal pregnancy. When compared to EP-hCG, GTD-hCG samples had decreased signals for tri- and tetra-antennary N-glycans. In terms of terminal epitopes, GTD-hCG had increased signals for sialylated structures, while LewisX antigens were of very minor abundance. hCG carries the same N-glycans throughout pregnancy but in different proportions. The N-glycan repertoire is more diverse than previously reported. Bisected and LewisX structures are potential targets for diagnostics. hCG isolated from pregnancy urine inhibits NK cell cytotoxicity in vitro at nanomolar levels and bisected type glycans have previously been implicated in the suppression of NK cell cytotoxicity, suggesting that hCG-related bisected type N-glycans may directly suppress NK cell cytotoxicity.

Supposed pituitary-production of human chorionic Gonadotropin induced by androgen deprivation therapy

ABSTRACT Introduction: The main cause of slightly elevated human chorionic gonadotropin (HCG) after successful treatment of male germ cell tumors is considered to be pituitary-derived HCG. It is well known that pituitary-derived HCG is frequently detected in postmenopausal women. We evaluated the status of serum HCG in men with elevated gonadotropins, which were induced by androgen deprivation therapy, using commercially available assays. Materials and Methods: We enrolled 44 patients with prostate cancer, who underwent luteinizing-hormone releasing hormone agonist treatment. We measured serum follicle-stimulating hormone (FSH), serum luteinizing hormone (LH), serum total HCG, serum free HCG-β subunit, and urine total HCG 3 times per patient, on the day of treatment initiation, the next day, and 3 months after. Results: On the day after treatment initiation, serum and urine HCG was detected in 61% and 73% of patients, respectively. Markedly strong correlations were observed between serum/urine HCG and FSH/LH. In particular, receiver operating characteristic curve analysis indicated excellent area under the curve (0.977, 95% confidence interval 0.951-1.003)) for serum HCG-detectable LH. At the cutoff value of 21.07 mIU/mL for serum HCG-detectable LH, the sensitivity and specificity were 96.7% and 95.3%, respectively. Serum HCG-β was not detectable at any times in any patients. Conclusions: Suggested pituitary-derived HCG can be frequently detected in patients with elevated gonadotropins, and there is a firm association between HCG detection and gonadotropin levels.

Supposed pituitary-production of human chorionic Gonadotropin induced by androgen deprivation therapy.

INTRODUCTION: The main cause of slightly elevated human chorionic gonadotropin (HCG) after successful treatment of male germ cell tumors is considered to be pituitary-derived HCG. It is well known that pituitary-derived HCG is frequently detected in postmenopausal women. We evaluated the status of serum HCG in men with elevated gonadotropins, which were induced by androgen deprivation therapy, using commercially available assays. MATERIALS AND METHODS: We enrolled 44 patients with prostate cancer, who underwent luteinizing-hormone releasing hormone agonist treatment. We measured serum follicle-stimulating hormone (FSH), serum luteinizing hormone (LH), serum total HCG, serum free HCG-ß subunit, and urine total HCG 3 times per patient, on the day of treatment initiation, the next day, and 3 months after. RESULTS: On the day after treatment initiation, serum and urine HCG was detected in 61% and 73% of patients, respectively. Markedly strong correlations were observed between serum/urine HCG and FSH/LH. In particular, receiver operating characteristic curve analysis indicated excellent area under the curve (0.977, 95% confidence interval 0.951-1.003)) for serum HCG-detectable LH. At the cutoff value of 21.07 mIU/mL for serum HCG-detectable LH, the sensitivity and specificity were 96.7% and 95.3%, respectively. Serum HCG-ß was not detectable at any times in any patients. CONCLUSIONS: Suggested pituitary-derived HCG can be frequently detected in patients with elevated gonadotropins, and there is a firm association between HCG detection and gonadotropin levels.

Massive hemorrhage from a unique old cesarean scar ectopic pregnancy with negative urine and serum ß-HCG: a rare case report.

Human chorionic gonadotropin (HCG) is an important indicator for the diagnosis of pregnancy. The authors report a unique case of cesarean scar ectopic pregnancy (CSEP) with negative urine and serum HCG levels, which was initially misdiagnosed as an intrauterine tumor despite the use of transvaginal ultrasound. Dilation and curettage was performed, which caused massive vaginal bleeding. Diagnostic hysteroscopy after uterine artery embolization and pathological examination of the surgical specimen confirmed the diagnosis of old CSEP. Postoperative follow-up showed that normal menstruation restarted 2 months later. This case reminds gynecologists and obstetricians the diagnosis of CSEP should be considered, especially when there is a mass at or near the surgical scar, regardless of the HCG level.

Emergency Medicine Myths: Ectopic Pregnancy Evaluation, Risk Factors, and Presentation.

BACKGROUND: Ectopic pregnancy (EP) is an important cause of morbidity and mortality in females of reproductive age. Proper diagnosis and treatment are critical, as complications such as rupture, hemorrhagic shock, and even death can occur. OBJECTIVE: EP is a condition emergency physicians are trained to detect, yet there are multiple myths concerning its evaluation and diagnosis. This article reviews several of these myths in order to improve emergency department (ED) evaluation and diagnosis. DISCUSSION: EP is a difficult diagnosis and may be missed on initial ED visit. While the diagnosis is often delayed simply due to very early presentations, it can also be missed because patients may not have all the same risk factors or demonstrate the same symptoms. They may also not demonstrate the same serum B-human chorionic gonadotropin levels and trends or have the same ultrasound findings at equivalent gestational ages. Some patients with early EP may have positive ultrasound findings with serum ß-hCG levels under a defined discriminatory zone (DZ). On the other hand, some patients with an early viable intrauterine pregnancy may have no visible findings on initial ultrasound, but have serum ß-hCG (quantitative) levels well above the DZ. Although rare, EP has even been demonstrated in women with negative urine ß-hCG tests or low serum ß-hCG levels. CONCLUSIONS: While EP may be a challenging diagnosis, understanding the myths surrounding EP may help emergency physicians consider it, even when patient risk factors, symptoms, or ED laboratory or imaging studies do not initially or easily define the diagnosis.

Negative ßhCG and Molar Pregnancy: The Hook Effect.

Molar pregnancy, included in gestational trophoblastic disease, is a benign pathology with ability to metastasize, usually occurring with excessively high ßhCG levels. Clinical scenario is usually a woman in extremes of reproductive age presenting with amenorrhoea, pain and vaginal blood loss; signs derived from high ßhCG levels may be present (hyperthyroidism, hyperemesis). Diagnosis is based on a positive pregnancy test - usually a qualitative urinary test. The limitation of this test results from its inability to become positive in presence of markedly high levels of ßhCG, saturating the antigens used - known as the 'hook effect'. With the widespread use of gynaecological ultrasound cases of molar pregnancy have been diagnosed in timely fashion. We describe a case referred as a degenerating fibroid, with a negative urinary pregnancy test. Transvaginal ultrasound was highly suggestive of molar pregnancy, which was confirmed with a quantitative ßhCG test, allowing for timely treatment. The importance of a high index of suspicion for this pathology is tremendous to avoid the devastating consequences of a delayed diagnosis.

A gravidez molar, incluída na doença gestacional do trofoblasto, é uma patologia benigna com capacidade de metastização, cursando com níveis excessivamente elevados de ßhCG. O quadro clínico traduz-se por amenorreia, dor e perda de sangue vaginal numa mulher frequentemente no extremo da idade reprodutiva, podendo estar presentes sinais decorrentes dos níveis de ßhCG (hipertiroidismo, hiperemese). O diagnóstico é histológico, e suspeitado por um teste de gravidez positivo, sendo normalmente realizado um teste urinário qualitativo. A limitação deste advém da incapacidade de se tornar positivo na presença de níveis exageradamente altos de ßhCG, que satura os antigénios utilizados ­ 'efeito hook'. Com a ecografia ginecológica os casos de gravidez molar têm sido diagnosticados mais atempadamente. Descrevemos um caso referenciado como um mioma degenerescente, com teste de gravidez urinário negativo. A ecografia transvaginal realizada foi altamente sugestiva de gravidez molar, confirmada com um teste quantitativo de ßhCG e permitindo tratamento atempado. A importância de um elevado índice de suspeição para esta patologia é fulcral para evitar as consequências devastadoras de um diagnóstico tardio.

Characterizing urinary hCGßcf patterns during pregnancy.

OBJECTIVE: Elevated concentrations of hCG beta core fragment (hCGßcf) are known to cause false-negative results in qualitative urine pregnancy test devices, but the pattern of urinary hCGßcf during normal pregnancy has not been well characterized. Here, we evaluate the relationship between urine hCG, hCGßcf, and hCG free ß subunit (hCGß) during pregnancy. DESIGN AND METHODS: Banked second trimester urine specimens from 100 pregnant women were screened for high concentrations of hCGßcf using a qualitative point-of-care device known to demonstrate false-negative results in the presence of elevated hCGßcf concentrations. Additional first and third trimester specimens from the same pregnancy were obtained from 10 women who generated negative/faint positive results, 5 women who generated intermediate positive results, and 10 women who generated strong positive results on the point-of-care device. Intact hCG, hCGßcf, hCGß, and specific gravity were quantified in these 75 specimens. RESULTS: Urinary hCGßcf concentrations were greater than intact hCG concentrations at all times. A strong correlation (r(2)=0.70) was observed between urine intact hCG and hCGßcf concentrations. A poor correlation was observed between specific gravity and intact hCG (r(2)=0.32), hCGß (r(2)=0.32), and hCGßcf (r(2)=0.32). The highest hCGßcf concentrations were observed between 10 and 16weeks gestation but individual women demonstrated very different patterns of hCGßcf excretion. CONCLUSIONS: Urine specimens with elevated hCGßcf are frequently encountered during pregnancy but hCGßcf excretion patterns are unpredictable. Manufacturers and clinicians must appreciate that hCGßcf is the major immunoreactive component in urine during pregnancy and must design and interpret qualitative urine hCG test results accordingly.

Estimating the hCGßcf in urine during pregnancy.

OBJECTIVE: Elevated urine concentrations of hCG beta core fragment (hCGßcf) are known to cause false negative qualitative point-of-care hCG test results, but limited information is available regarding urine hCGßcf. In this study, we evaluate the relationship between serum and urine hCG concentrations and the frequency of elevated urine hCGßcf concentrations. DESIGN AND METHODS: Paired serum and urine specimens were obtained from 60 women at various stages of pregnancy and hCG was measured using the Abbott Architect and Roche Cobas e602 assays. Urine specimens with the greatest difference in urine hCG concentrations between these two instruments were tested using a qualitative point-of-care device and hCGßcf was quantified using LC-MS/MS. RESULTS: Urine hCG concentrations were lower than serum and the magnitude of the difference depended on whether the hCG assay detected hCGßcf. Elevated hCGßcf concentrations (>280,000pmol/L) were observed in 12% of specimens from an unselected patient population. There was a significant correlation (r=0.97; p<0.0001) between the difference (Roche hCG-Abbott hCG) and the hCGßcf concentration as measured by LC-MS/MS (Roche-Abbott difference IU/L=(hCGßcf (pmol/L)∗0.131+656)). CONCLUSIONS: A correlation exists between serum and urine hCG concentrations but this correlation is variable. hCGßcf concentrations can be estimated using two automated assay reagent platforms that differ in their recognition of hCGßcf.

Twin tubal pregnancy: A large unruptured ectopic pregnancy.

Twin ectopic pregnancy is a rare occurrence, with an estimated incidence of 1 in 20 000 spontaneous pregnancies. We describe a case of unilateral twin ectopic pregnancy in which the gravid fallopian tube showed no signs of tubal rupture despite marked tubal distension. A 25-year-old woman presented with clinical features suggestive of large right-sided tubal ectopic pregnancy. Serum ß-human chorionic gonadotropin was 10 800 IU/mL. Laparotomy revealed markedly distended right fallopian tube. There was no hemoperitoneum. The tube contained twin gestational sacs. The crown-rump length of the embryos was 2 cm. The ectopic gestation was thus unique, in that despite marked tubal distension, the trophoblastic invasion was not significant to cause tubal rupture. There may be a role for medical management based on individual gestational sac size in selected cases of twin tubal pregnancy in which there is no evidence of hemoperitoneum.

Evaluation of a semi-quantitative pregnancy device for susceptibility to interference caused by hCGßcf.

OBJECTIVE: Previous work has documented the ability of the Clearblue Advanced Test with Weeks Estimator, a new over-the-counter (OTC) urine hCG device, to accurately estimate weeks since ovulation in early pregnancy. In this study, the performance of this device in more advanced pregnancy was assessed. METHODS: The Clearblue Advanced Test with Weeks Estimator device was used to test solutions containing purified intact hCG and hCGßcf at concentrations consistent with early, middle and late pregnancy. Urine samples from three normal pregnant patients 9-13 weeks of gestation and from a patient 12 weeks of gestation known to generate false negative results on qualitative urine test devices due to excess hCGßcf were also evaluated. RESULTS: The Clearblue Weeks Estimator device gave expected results using solutions containing purified intact hCG and hCGßcf at concentrations observed throughout pregnancy. The device generated expected results using urine from three of four patients tested between 9 and 13 weeks of gestation. However, when urine from a patient with elevated concentrations of hCGßcf was used, the device correctly indicated pregnancy although the estimate for the date was incorrect. CONCLUSION: This device gave expected "pregnant" results using all samples tested. However, the "Weeks Estimator" should be interpreted with caution when used by patients after seven weeks of pregnancy.

Quantitative analysis of total ß-subunit of human chorionic gonadotropin concentration in urine by immunomagnetic reduction to assist in the diagnosis of ectopic pregnancy.

BACKGROUND: The initial diagnosis of ectopic pregnancy depends on physical examination, ultrasound, and serial measurements of total ß-subunit of human chorionic gonadotropin (hCGß) concentrations in serum. The aim of this study was to explore the possibility of using quantitative analysis of total hCGß in urine rather than in serum by immunomagnetic reduction (IMR) assay as an alternative method to diagnose an ectopic pregnancy. METHODS: We established a standard calibration curve of IMR intensity against total hCGß concentration based on standard hCGß samples, and used an IMR assay to detect total hCGß concentrations in the urine of pregnant women with lower abdominal pain and/or vaginal bleeding. The final diagnosis of ectopic pregnancy was based on ultrasound scans, operative findings, and pathology reports. In this prospective study, ten clinical samples were used to analyze the relationship of total hCGß IMR signals between urine and serum. Furthermore, 20 clinical samples were used to analyze the relationship between urine IMR signals and serum levels of total hCGß. RESULTS: The calibration curve extended from 0.01 ng/mL to 10,000 ng/mL with an excellent correlation (R(2)=0.999). In addition, an excellent correlation of total hCGß IMR signals between urine and serum was noted (R(2)=0.994). Furthermore, a high correlation between urine IMR signals and serum levels of total hCGß was noted (R(2)=0.862). CONCLUSION: An IMR assay can quantitatively analyze total hCGß concentrations in urine, and is a potential candidate for point-of-care testing to assist in the diagnosis of ectopic pregnancy.

Using a simulation model to assess risk of false negative point-of-care urinary human chorionic gonadotropin device results due to high-dose hook interference.

BACKGROUND: It is unclear if the point-of-care (POC) Clinitest hCG device is subject to high-dose hook interference from physiological concentrations of intact human chorionic gonadotropin (hCG), ß-core fragment of hCG (hCGßcf), and hCG free ß-subunit (hCGß) found in urine during pregnancy. We used a simulation model to address this question and related our findings to our institution's pregnant population in order to assess risk for potential false-negative hCG results. METHODS: The expected distribution of days relative to ovulation during routine POC hCG testing was estimated from 182 patients. Clinitest-Clinitek Status hCG device susceptibility to high-dose hook interference from hCG variants and potential risk of false-negative results as it relates to this population were evaluated by testing increasing concentrations of hCG, hCGßcf, hCGß as well as urine simulating physiological hCG, hCGßcf and hCGß concentrations expected during early pregnancy (≤44 days post-ovulation). RESULTS: The Clinitest-Clinitek Status hCG device exhibited high-dose hook interference from hCGßcf alone, but not from hCG, hCGß, or simulated physiological urinary concentrations of combined hCG, hCGßcf and hCGß expected during early pregnancy. The majority of our patient population had urinary hCG testing conducted during early pregnancy. CONCLUSION: The Clinitest-Clinitek Status hCG device is unlikely to exhibit false-negative urinary hCG results due to high-dose hook interference for women in early healthy pregnancy, although additional studies are necessary to determine potential risk in other patient populations. Visual interpretation of POC urinary hCG device results is an important failure mode to consider in risk analyses for erroneous urinary hCG device results.

Direct analysis of hCGßcf glycosylation in normal and aberrant pregnancy by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

The analysis of human chorionic gonadotropin (hCG) in clinical chemistry laboratories by specific immunoassay is well established. However, changes in glycosylation are not as easily assayed and yet alterations in hCG glycosylation is associated with abnormal pregnancy. hCGß-core fragment (hCGßcf) was isolated from the urine of women, pregnant with normal, molar and hyperemesis gravidarum pregnancies. Each sample was subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) analysis following dithiothreitol (DTT) reduction and fingerprint spectra of peptide hCGß 6-40 were analyzed. Samples were variably glycosylated, where most structures were small, core and largely mono-antennary. Larger single bi-antennary and mixtures of larger mono-antennary and bi-antennary moieties were also observed in some samples. Larger glycoforms were more abundant in the abnormal pregnancies and tri-antennary carbohydrate moieties were only observed in the samples from molar and hyperemesis gravidarum pregnancies. Given that such spectral profiling differences may be characteristic, development of small sample preparation for mass spectral analysis of hCG may lead to a simpler and faster approach to glycostructural analysis and potentially a novel clinical diagnostic test.

Significant immunohistochemical expression of human chorionic gonadotropin in high-grade osteosarcoma is rare, but may be associated with clinically elevated serum levels.

Survival rates have plateaued at 70% for osteosarcoma. Proteins ectopically produced by malignant tumors may provide insight into new therapeutic targets. Osteosarcomas secreting human chorionic gonadotropin (hCG) have been suggested to have a worse prognosis. We examined the frequency of expression of ß-subunit of hCG (ß-hCG) in pretreatment osteosarcoma biopsies, and asked if it was associated with various clinical prognostic parameters, and the development of metastases. We subjected 51 pretreatment biopsies of high-grade osteosarcoma, from 51 patients, to ß-hCG immunohistochemistry. In 19 of these patients, postchemotherapy metastatic biopsies also were examined for ß-hCG expression. Clinical information (patient age, sex, survival status, and serum hCG in females only), and tumor characteristics (site, size, and presence of metastases) were recorded. The ß-hCG positive and negative biopsies were separated and compared. Of 49 interpretable pretreatment biopsies, 28 (57%) showed positive cytoplasmic ß-hCG expression: 27 with sparse positivity (1% of tumor cells) and 1 with frequent positivity (10% of tumor cells). The patient with frequent ß-hCG positivity in her pretreatment biopsy had elevated serum hCG (88.2 mIU/mL) at diagnosis, decreasing to undetectable following chemotherapy and definitive resection. There was no difference in clinical parameters or rate of metastasis between ß-hCG positive versus negative groups. Expression of ß-hCG may be seen in high-grade osteosarcoma, but frequent ß-hCG immunohistochemical expression by tumor cells, associated with clinically elevated serum ß-hCG, is rare. Recognition that some nongerm cell tumors may produce ß-hCG can prevent confusion with malignancies containing neoplastic syncytiotrophoblast cells, including germ cell and trophoblastic tumors.

Krukenberg tumor presenting as back pain and a positive urine pregnancy test: a case report and literature review.

A Krukenberg tumor is a rare and potentially deadly cause of elevated serum ß-hCG as part of a paraneoplastic syndrome. This study aims to describe the unusual case of a 36-year-old woman that presented to the Emergency Department (ED) with back pain and a positive urine pregnancy test. Assessment revealed no intrauterine pregnancy and a small left ovarian cyst. Further investigation showed moderately differentiated gastric adenocarcinoma with distant metastases to the spine. The patient died less than 3 months after her first presentation to the ED. Paraneoplastic syndrome, albeit rare, should be considered in the differential diagnosis of elevated ß-hCG due to the high mortality associated with Krukenberg tumors.

[Point-of-care testing in preclinical emergency medicine]. / Point-of-care-Testung in der präklinischen Notfallmedizin.

BACKGROUND: Measurement of biological signals directly at the patient (point-of-care testing, POCT) is an established standard in emergency medicine when test results are needed quickly and within a reliable time frame or if external testing requires a disproportionate effort. OBJECTIVES: Currently, the rapid test for ß-HCG in urine and POCT measurement of lactate, blood gases, cardiac tropinin, haemoglobin, and hematocrit are well established in emergency medicine. POCT of copeptin, fatty acid-binding proteins (FABP), procalcitonin, coagulation values, natriuretic peptides, D-dimer, and toxicological substances are of future interest. In this article, the appropriate use of point-of-care testing in prehospital emergency medicine is discussed. RESULTS: Application of POCT is dependent of the underlying conditions, the availability of appropriate devices, and of suitable reference methods in a central laboratory. In addition, economical and quality aspects play an important role. CONCLUSION: In emergency departments, POCT is currently developing into a standard measuring method for a number of markers because hospital laboratories are increasingly being merged and consequently reduce their emergency-analytic services. In countries with a high density of hospitals, however, preclinical POCT should be reduced to the minimum necessary.