Hysteroscopic Intact Removal of an Angular Pregnancy with a 5Fr Electrode.

STUDY OBJECTIVE: Angular pregnancy is a rare and life-threatening condition in which the embryo is implanted in the lateral angle of the uterine cavity, medial to the uterotubal junction and round ligament. Angular pregnancy is associated with a high risk of uterine rupture of about 23% [1]. No consensus has been achieved regarding the diagnostic and therapeutic approach of angular pregnancy [2]. Thus, the aim of this study was to report a case of hysteroscopic treatment of an angular pregnancy in a 34-year-old women. DESIGN: Step-by-step video presentation of the surgical treatment (Canadian Task Force classification III). SETTING: Department of Neuroscience, Reproductive Sciences and Dentistry, School of Medicine, University of Naples Federico II, Naples, Italy. PATIENT: A 34-year-old woman. Written informed consent was obtained from the patient. INTERVENTION: Hysteroscopy. MEASUREMENTS AND MAIN RESULTS: A 34-year-old woman was admitted to our Department with pelvic pain at 6 weeks of gestation. ß-Human chorionic gonadotropin (ß-hCG) was 5331 mIU/mL. The transvaginal ultrasound showed a gestational sac of 15 × 11 mm in the left uterine angle of an embryo without cardiac activity. The woman opted for a conservative approach with multiple-dose methotrexate [3]. Five days later the ß-hCG increased to 7589 mIU/mL with no regression of pregnancy at the transvaginal ultrasound. Therefore, a surgical approach was offered to the patient [4,5]. Laparoscopy showed normal salpinges, whereas hysteroscopy identified the gestational sac in the left uterine angle. A 5Fr bipolar electrode was used to open the gestational capsular decidua. The chorionic villi were progressively separated from the implantation site. Using grasping forceps we removed the specimen for histologic examination. Histologic examination confirmed the diagnosis of angular pregnancy. On the second postoperative day ß-hCG was 1131 mIU/mL, and the patient was discharged the day after. At the 1-month follow-up visit, ß-hCG and transvaginal ultrasound were negative for pregnancy. The office hysteroscopy showed an empty uterine cavity at 3-months' follow-up. CONCLUSION: Our case shows that hysteroscopy may be used as a diagnostic and therapeutic tool for angular pregnancy, providing a unique image of the intact removal of the gestational sac.

Evaluation of the Efficacy of Ultrasound-Guided Local Lauromacrogol Injection Combined with Aspiration for Cesarean Scar Pregnancy: A Novel Treatment.

OBJECTIVES: To evaluate the efficacy of ultrasound-guided local lauromacrogol injection combined with aspiration for treating cesarean scar pregnancy (CSP). METHODS: From July 2016 to December 2016, 18 patients diagnosed with CSP were treated with ultrasound-guided local lauromacrogol injection combined with aspiration. Clinical data and outcome were analysed. RESULTS: All patients were treated successfully. The amount of bleeding ranged between 10 and 50 mL. The duration of hospitalization ranged between 2 and 11 days. Serum ß-human chorionic gonadotropin (ß-hCG) decreased to the nondetectable level within 19-41 days. Menstruation recovery occurred after 10-24 days of normalization of serum ß-hCG level. Reproductive functions were preserved, and there were no untoward effects or complications. CONCLUSIONS: Ultrasound-guided local lauromacrogol injection combined with aspiration is an effective CSP therapy, as it was associated with a high success rate, short hospitalization and fast recovery. However, its wider application and popularization have to be validated on a larger patient population affected by CSP.

Efficient isolation of encoded microparticles in a degassed micromold for highly sensitive and multiplex immunoassay with signal amplification.

Multiplex detection of low-abundance protein biomarkers in biofluids can contribute to diverse biomedical fields such as early diagnosis and precision medicine. However, conventional techniques such as digital ELISA, microarray, and hydrogel-based assay still face limitations in terms of efficient protein detection due to issues with multiplexing capability, sensitivity, or complicated assay procedures. In this study, we present the degassed micromold-based particle isolation technique for highly sensitive and multiplex immunoassay with enzymatic signal amplification. Using degassing treatment of nanoporous polydimethylsiloxane (PDMS) micromold, the encoded particles are isolated in the mold within 5 min absorbing trapped air bubbles into the mold by air suction capability. Through 10 min of signal amplification in the isolated spaces by fluorogenic substrate and horseradish peroxidase labeled in the particle, the assay signal is amplified with one order of magnitude compared to that of the standard hydrogel-based assay. Using the signal amplification assay, vascular endothelial growth factor (VEGF) and chorionic gonadotropin beta (CG beta), the preeclampsia-related protein biomarkers, are quantitatively detected with a limit of detection (LoD) of 249 fg/mL and 476 fg/mL in phosphate buffer saline. The multiplex immunoassay is conducted to validate negligible non-specific detection signals and robust recovery rates in the multiplex assay. Finally, the VEGF and CG beta in real urine samples are simultaneously and quantitatively detected by the developed assay. Given the high sensitivity, multiplexing capability, and process simplicity, the presented particle isolation-based signal amplification assay holds significant potential in biomedical and proteomic fields.

The efficacy of different treatments for type 2 cesarean scar pregnancy.

OBJECTIVE: To study the efficacy of 3 treatment options for type 2 cesarean scar pregnancy (CSP) and establish an optimal treatment strategy for type 2 CSP. DESIGN: Retrospective cohort study. SETTING: A tertiary hospital. PATIENTS: The study examined 160 women with type 2 CSP. INTERVENTIONS: Ultrasound-guided vacuum aspiration after local injection of lauromacrogol, ultrasound-guided vacuum aspiration after uterine artery embolization (UAE), and transabdominal resection or hysteroscopy combined with laparoscopic resection. MAIN OUTCOME MEASURES: The success rates, duration of hospitalization, hospitalization cost, amount of blood loss, recovery time, and menstruation resuming after recovery. RESULTS: The success rates of the UAE, lauromacrogol, and surgical groups were 87.1%, 92.5%, and 95.5%, respectively, with no significant differences. The cost and duration of hospitalization in the lauromacrogol group were significantly lower than those in the UAE and surgical groups. Analysis of the causes of treatment failure revealed a significant difference in the gestational age. The area under the receiver operating characteristic curve was 0.660 (95% confidence interval, 0.533-0.788). When the gestational age was 48.5 days, Youden index was the highest. Furthermore, when the diagnostic thresholds were selected as 49, 56, and 63 days of pregnancy, the corresponding areas under the receiver operating characteristic curve were 0.652, 0.541, and 0.510, respectively. CONCLUSION: Ultrasound-guided vacuum aspiration after local injection of lauromacrogol is recommended for patients with type 2 CSP at <49 days of gestation. Laparotomy or laparoscopy combined with hysteroscopy is suitable for patients with gestation of >49 days, especially for those with >56 days of gestation.

Kinetic analysis of a human chorionic gonadotropin-beta epitope-paratope interaction.

Kinetics of protein-protein or ligand-ligate interaction has predominantly been studied by optical spectroscopy (particularly fluorescence) and surface plasmon resonance biosensors. Almost all such studies are based on association kinetics between ligand-ligate and suffer from certain methodological and interpretational limitations. Therefore, kinetic analyses of dissociation data of such interactions become indispensable. In the present investigation, the radiolabeled human chorionic gonadotropin-beta ((125)IhCGbeta) was employed as a probe and nitrocellulose (NC) as a solid support to immobilize monoclonal antibody (MAb) G(1)G(10).1. The NC-G(1)G(10).1-(125)IhCGbeta complex (NC(com)) was prepared and the dissociation of radiolabeled hCGbeta was carried out in the presence of excess unlabeled ligate. From the experimental dissociation data under varying ionic strength, dissociation constants (k(- 1)), association constants (k(+1)) and affinity constants (k(a)) were calculated. The values obtained were utilized in exploring the amino acid residues constituting an epitopic region of hCGbeta involved in interaction with the complementary paratope on MAb G(1)G(10).1. Kinetic data of the present study supported our recently published findings [using single step-solid phase radioimmunoassay (SS-SPRIA)] that the core region of hCGbeta epitope consists of Arg (94,95) and Asp (99) while a Lys (104) and a His (106) are in proximity to the core epitopic region. Based on the results of present investigation, we conclude that dissociation kinetics coupled with SS-SPRIA unequivocally provides considerable insight into the study of ligand-ligate interactions and epitope analysis.

Biochemical markers for the prediction of preterm delivery.

Preterm delivery is the largest contributor to perinatal morbidity and mortality throughout the world. In the United States, nearly 1 in every 8 infants is born prematurely. Although a portion of these births are indicated preterm deliveries, the frequency of spontaneous preterm birth has remained largely constant over the past 50 years.

Simplifying the preevacuation testing strategy for patients with molar pregnancy.

OBJECTIVE: To review our institution's preevacuation testing strategy for suspected molar pregnancy to determine whether a simplified approach might be indicated. STUDY DESIGN: Patients diagnosed with molar pregnancy from 1999 to 2004 were identified. Clinical data were retrospectively extracted from medical records. RESULTS: One hundred fifty-eight women diagnosed (mean age, 24 years) underwent dilatation and curettage. Molar pregnancy was suspected at presentation in 111 (70%); 47 (30%) cases were presumed miscarriages, and the diagnosis was confirmed only after histologic evaluation of the specimen. Initial testing included complete blood count (CBC) (87%), liver function tests (LFT) (63%), thyroid-stimulating hormone (TSH) level (72%), clotting function studies (26%) and chest radiograph (84%). One patient with right upper quadrant pain had elevated LFTs and a coagulopathy that resolved after evacuation. One woman with a palpably enlarged goiter and elevated TSH level was diagnosed with thyroid carcinoma. No chest radiograph demonstrated metastatic disease. CONCLUSION: We advocate a simplified approach to preevacuation testing for suspected molar pregnancy that includes a CBC and blood type with antibody screen. Clinical assessment should prompt additional evaluation in the rare patient with suspicious signs and symptoms.

Evaluation of cytotoxic and tumor targeting capability of (177)Lu-DOTATATE-nanoparticles: a trailblazing strategy in peptide receptor radionuclide therapy.

OBJECTIVE: We propose an innovative strategy of nanoparticle-mediated-peptide receptor radionuclide therapy (PRRT) employing PLGA-nanoparticles together with anti-ß-hCG antibodies that can protect kidneys from radiation damage while simultaneously enhancing its tumor targeting and cytotoxic ability for somatostatin receptor (SSR) positive tumors. METHODS: PEG-coated-(177)Lu-DOTATATE-PLGA-nanoparticles (PEG-LuD-NP) were formulated and characterized. In vitro toxicity of these particles was tested on human glioblastoma cell line U87MG over a radiation dose range of 19-78 Gy, using MTT assay and flow cytometry. To further enhance cytotoxicity and test the feasibility of active tumor targeting, apoptosis-inducing anti-ß-hCG monoclonal antibodies were employed in vitro, after confirming expression of ß-hCG on U87MG. In vivo tumor targeting ability of these particles, in comparison to uncoated particles and un-encapsulated (177)Lu-DOTATATE, was assessed by intravenous administration in tumor-induced wistar rats. Rats were first imaged in a gamma camera followed by euthanasia for organ extraction and counting in gamma counter. RESULTS: The particles were spherical in shape with mean diameter of 300 nm. Highest cytotoxicity that could be achieved with PEG-LuD-NP, on radio-resistant U87MG cells, was 35.8 % due to complex cellular response triggered by ionizing radiation. Interestingly, synergistic action of antibodies and PEG-LuD-NP doubled the cytotoxicity (80 %). PEG-LuD-NP showed the highest tumor uptake (4.3 ± 0.46 % ID/g) as compared to (177)Lu-DOTATATE (3.5 ± 0.31 %) and uncoated-(177)Lu-DOTATATE-nanoparticles (3.4 ± 0.35 %) in tumor-inoculated wistar rats (p < 0.001). Renal uptake/retention was decreased 3-4 folds with these particles, resulting in the highest tumor-to-kidney ratio (8.58; p < 0.01) while tumor-to-liver and tumor-to-bone ratios were comparable to un-encapsulated-drug. CONCLUSION: Nanocarrier-mediated-PRRT is an effective way of targeting SSR positive tumors for enhanced cytoxicity and reduced renal radiation dose associated with conventional PRRT. To our knowledge of literature, this is the first study to establish in vitro and in vivo efficacy profile of nanoparticles in PRRT providing a stepping-stone for undergoing and future research endeavors in the direction of abating associated radiation concerns of radionuclide therapy and may offer a paradigm shift in PRRT strategy.

Effects of a beta-human chorionic gonadotropin subunit immunogen administered in aqueous solution with a novel nonionic block copolymer adjuvant in patients with advanced cancer.

The clinical and immunological effects of a vaccine consisting of CTP37, a synthetic peptide corresponding to the COOH-terminal peptide (CTP) of beta-human chorionic gonadotropin (beta-hCG) conjugated to diphtheria toxoid, combined with CRL 1005, a novel synthetic nonionic block copolymer adjuvant, were examined. Twenty-one patients with metastatic, nontrophoblastic cancers received up to four immunizations by i.m. injection of a fixed dose of CTP37 and escalating doses of CRL 1005. Doses of CRL 1005 adjuvant as high as 75 mg were administered with 1 mg of CTP37 without evidence of significant local or systemic toxicity. Immunizations resulted in the production of IgG antibody to beta-hCG. CRL 1005 doses of 3-25 mg appeared to be optimal for antibody induction. Immunizations also resulted in increases in the cellular response of peripheral blood mononuclear cells (PBMCs) to the unconjugated CTP, hCG, and diphtheria toxoid. Responding PBMCs specifically secreted the TH1-associated cytokines IFN-gamma and interleukin (IL)-2 as well as the TH2-associated IL-5 and IL-10. Increased expression of IFN gamma and IL-5 mRNAs by PBMCs 4 h after immunization was also observed. CRL 1005 administered with CTP37 in aqueous solution is well tolerated. The CTP37-CRL 1005 subunit vaccine has the capacity to stimulate potentially beneficial humoral and cellular immune responses in patients with advanced cancer.

Collection of buccal cell DNA in seventh-grade children using water and a toothbrush.

We developed a simple and effective method for collecting a large quantity of buccal cell DNA in school-based studies of seventh-grade and older children. Seventh-grade students at schools in Wuhan, China brushed each buccal surface with a soft toothbrush and then rinsed with 10 ml of water. We added 5 ml of 99% ethanol to preserve the sample. Among 1563 samples transported at room temperature over 1 week and then stored for 13-14 months at -70 degrees C before extraction, using a modified Gentra Puregene protocol, the median total DNA yield was 108 microg, range of 14 to 416 microg. We assayed every 20th sample (n = 77) for NAT2 by the PCR, and all samples gave a 1093-bp product. From the 1563 samples, we obtained a result for single nucleotide polymorphisms in the interleukin-13 gene (at +2044) by RFLP-PCR on 98.8% and in the promoter of the myeloperoxidase gene (at -463) by real-time PCR on 99.7%. A water-rinse method, that we used among 12th-grade students in Southern California, gave a lower total DNA yield than the toothbrush rinse (median of 17 microg) and a slightly reduced ability to generate a PCR product. However, 26 of 27 water-rinse samples gave a result for two genes, albumin and CYP1A1, using real-time PCR methods. We did not quantify human, versus bacterial, DNA in our samples. However, given the amounts of total DNA required for genotyping, a sample with the median yield of 108 microg should suffice for approximately 2160 genotypes by RFLP-PCR methods or five times as many by real-time PCR. We recommend the toothbrush-rinse method, combined with a modified Gentra Puregene DNA extraction protocol, for large-scale, in-person collections of buccal cell DNA in children. The method requires only inexpensive, readily available materials and produces a large quantity of high-quality DNA for PCR analyses.

ELISA solid phase: stability and binding characteristics.

Stability of solid-phase antibody (SP-Ab) was investigated using polystyrene surface coated, by simple physical adsorption, with a rabbit antibody against the beta subunit of human chorionic gonadotropin. SP-Ab stored wet in 0.1 M Tris-HCl buffer, pH 7.2, or in 10% ammonium sulphate solution in the Tris buffer did not lose any activity upto the entire period of the study, i.e., 95 days at 6 degrees C and room temperature and 53 days at 40 degrees C. However, SP-Ab lost some activity upon drying and decrease in activity continued when air-dried SP-Ab was stored at any of these temperatures. Secondary coating of SP-Ab with sucrose or lactose before drying offered only partial protection to the activity of SP-Ab. Using radiolabeled antibody it was found that the loss of activity was not caused by desorption of the antibody; rather it could be the result of surface denaturation of the molecules. Binding of the antibody to polystyrene was so strong that week-long storage of SP-Ab in 6 M urea or 0.1% acetic acid could not bring about desorption by more than 1.8%. It is recommended that SP-Ab should be stored in a buffer and should not be allowed to dry.

High frequency of in situ hybridization on thin plastic sections of human placenta with a cDNA probe for beta hCG.

We describe two different techniques with plastic embedding in in situ hybridization histochemistry (ISHH). Their applicability was demonstrated by use of human placenta of the tenth gestational week and a tritium-labeled cDNA probe for the beta-subunit of hCG. In the first method, ISHH was performed on whole pieces of tissue (en bloc ISHH) pretreated with a weak acid solution, embedded in methacrylate, and sectioned at 3 microns for autoradiography. In the second technique, en bloc ISHH was carried out on tissue pre-treated with the weak acid and thereafter with detergent to further facilitate probe penetration. An acrylic resin was used for embedding, and section thickness was reduced to 1 microns. With both techniques, beta hCG cDNA/mRNA hybrids were localized exclusively to the syncytiotrophoblast (ST), in agreement with a previous study using sections of frozen placentas for hybridization to the same probe. However, owing to the higher resolution of the plastic sections the reliability of this localization was greatly increased. The number of autoradiographic grains over the acrylic resin 1-microns sections was found to be considerably higher than that over the methacrylate 3-microns sections. This study showed that treatment of tissue with detergent before en bloc ISHH, with subsequent embedding in acrylic resin and sectioning at 1 microns, gives high resolution in combination with a high signal-to-noise ratio after autoradiography. As the acrylic resin permits cutting of ultrathin sections, the results suggest that the technique may become useful for ISHH studies at the subcellular level.

A comparison of serial quantitative serum and urine tests in early pregnancy.

Serial urinary beta human chorionic gonadotropin (beta-hCG) immunoassay was performed on 60 patients during early pregnancy. Results were expressed as tube dilutions positive and were compared with quantitative serum beta human chorionic gonadotropin radioimmunoassay (beta-hCG-RIA) values. The parallel rise of human chorionic gonadotropin (hCG) measured by serum beta-hCG-RIA and a macroflocculation beta-specific urinary immunoassay in early pregnancy was confirmed. The ability of each to predict abortion prior to the onset of patient symptoms or clinical signs of disturbed gestation was quantitated. Results were expressed for each method according to standard regression lines or doubling time for individuals. Utilizing either statistical method, urinary testing was as accurate as serum testing for the prediction of normal pregnancy (about 90% for each). Serum testing was 78% correct in predicting abortion; urinary testing was 63% accurate.

[Infantile and maternal choriocarcinoma]. / Choriocarcinome infantile et maternel.

Infantile or congenital choriocarcinoma is a very uncommon complication of gestational choriocarcinoma. We report such a case with fatal outcome in a 3-week-old newborn, admitted for a hemorrhagic syndrome. Lungs, liver and brain masses were discovered and suggested an angiomatous process. The diagnosis was made later on gingival biopsy with necropsic confirmation. The mother's B-HCG level was elevated. She had asymptomatic pulmonary nodules and a uterine mass. This case report highlights characteristic but non specific clinical findings leading to the diagnosis. Chemotherapy must be undertaken as soon as possible to be effective. It is also necessary to assay maternal serum B-HCG when infantile choriocarcinoma is disclosed.

Prenatal sonographic features of fetuses in trisomy 13 pregnancies. IV.

Prenatal ultrasound is a powerful tool to detect structural abnormalities associated with the fetuses in trisomy 13 pregnancies. This article provides a comprehensive review of the prenatal sonographic markers of trisomy 13 in the first trimester, including fetal nuchal translucency thickness, fetal heart rate, fetal nasal bone, fetal tricuspid regurgitation, ductus venous flow, fetal crown-rump length, fetal trunk and head volume, fetal frontomaxillary facial angle, gestational sac volume and umbilical cord diameter, along with biochemical markers such as maternal serum free beta-human chorionic gonadotropin, maternal serum pregnancy-associated plasma protein-A, maternal serum placental growth factor, and the fetal and total cell-free DNA concentration in the maternal circulation.

Effect of variations in globulin concentrations on serum radioimmunoassay results, as exemplified by choriogonadotropin.

Variations in serum protein concentrations can affect results of radioimmunoassay of human choriogonadotropin involving either polyethylene glycol or double-antibody methods of separation. Relative to the maximal binding of choriogonadotropin in the presence of normal (30 g/L) globulin concentrations, globulin concentrations of 10 to 60 g/L were associated with maximal binding ranging from 84.9 to 115.6% in the polyethylene glycol system, and from 109.9 to 92.5% in the double-antibody system. This effect was quantitatively much the same throughout the standard curve in the polyethylene glycol system, but diminished with the higher standard concentrations in the double-antibody system. In assays involving polyethylene glycol separation, an increase in globulin was associated with a linear increase in nonspecific binding, but no detectable effect of globulin on nonspecific binding was observed in the double-antibody system. Determination and compensation for nonspecific binding of individual samples (rather than for mean assay nonspecific binding) eliminated the globulin effect in the polyethylene glycol system but not in the double-antibody system.

The measurement of the peptide hormones in saliva. 2. Chorionic gonadotropin subunits in pregnancy.

The alpha and beta subunits of the human chorionic gonadotropin (HCG) were measured during pregnancy in the saliva of a first group (no. = 23) and in the serum and saliva of a second group (no. = 25). Only ten pregnant women had normal gestation, the remaining 39 being diagnosed as miscellaneous pathologies, the threatened (or impending) abortion being the most frequent (no. = 19). The mean serum levels of alpha-HCG increase from 75.97 +/- 18.59 ng/ml (X +/- SEM) during the first trimester to 341.98 +/- 65.09 ng/ml during the third trimester of the gestation. The mean salivary concentration of the alpha HCG seems unmodified during pregnancy (approximately 10 ng/ml), but large interindividual variations were observed (limits 0.10-28.78 ng/ml) possibly due to the non-homogeneity of the investigated groups. The presence of the beta-HCG subunit in saliva could not be assessed, the great part of the values being aggregated around the sensitivity limit of the RIA technique (0.2 ng/ml). The physiological significance of the presence of the HCG and of its subunits in saliva as well as the ways to elucidate the possible selective role of the blood-saliva barrier are discussed.

An unusual localisation of Kaposi's sarcoma: the external auditory canal.

The epidemic form of Kaposi's sarcoma is associated with human immunodeficiency virus infection. Cutaneous and mucosal manifestations are frequently reported in the ENT sphere, mostly involving the oral cavity. The external and middle ear are only rarely concerned with only one case of a mastoid lesion without extension to the external auditory canal (EAC) being reported to this day. The present article describes the first case of involvement of the EAC with extension to adjacent structures. This patient presented other Kaposi lesions and had been treated by systemic hormonal therapy. Thereafter local injection of a cytotoxic agent was given without effect. Finally, radiotherapy resulted in a 50% regression of the tumour mass. The epidemiologic factors and therapeutic modalities with their results are described.