Failure of hCG/LH receptors to stimulate the transmembrane effector adenylyl cyclase in human endometrium

The functional significance of the endometrial hCG/ LH receptors has been related to a rapid release of prostaglandins. However, as compared to gonads and myometrium, in-endometrium mechanisms of transmembrane signalling of the hCG/LH receptors are probably not conventional and remain unclear. Here we investigated, in vivo, the potential of hCG to interact with, and stimulate the membrane effector enzyme, adenylyl cyclase (AC), in human endometrium. Hormonal and nonhormonal activation of AC was tested in membrane fractions prepared from endometrial biopsies obtained from patients undergoing evaluation cycles for hormone replacement therapy (HRT) and controlled ovarian hyperstimulation (COH). AC activity was determined by the direct conversion of the substrate ATP into cAMP under unstimulated conditions and in the presence of the non-hormonal activators guanyl nucleotide and forskolin. Also AC activity was tested in the presence of hCG under conditions allowing maximal enzyme stimulation. Isoproterenol and prostaglandin E2 (PGE2) were included for comparison. Immunoblot analyses demonstrated the presence of hCG/LH receptors and Gsα protein and other members of the G protein family in the membrane fractions. Endometrial membranes also exhibited high levels of AC activity compared to luteal membranes used as control. Stimulation by GMP-P(NH)P alone was 196 ± 63 (n = 8) (pmol/mg/ min ± SD). Neither hCG nor isoproterenol showed stimulation of endometrial AC (210 ± 65, and 197 ± 53, respectively;n = 66 assays). But PGE2 stimulated the enzyme system significantly (264 ± 63, p < 0.05;n = 66 assays). These data show that membrane fractions from human endometrium express all the AC system components, namely, hCG/LH receptors, Gsα protein and AC;however, hCG does not stimulate the endometrial AC system. Our data indicate that, in great contrast to gonadal receptors, endometrial hCG/ LH receptors are not coupled to the transmembrane AC effector. The well known release of eicosanoids in response to hCG suggests that these receptors are functional in human endometrium but throughout a signalling system different from AC. This enzyme is certainly coupled to and directly activated by eicosanoids and other embryonic signals.

Inflammatory bowel disease: A descriptive study of 716 local Chilean patients

AIM: To demographically and clinically characterize inflammatory bowel disease(IBD) from the local registry and update data previously published by our group.METHODS: A descriptive study of a cohort based on a registry of patients aged 15 years or older who were diagnosed with IBD and attended the IBD program at Clínica Las Condes in Santiago, Chile. The registry was created in April 2012 and includes patients registered up to October 2015. The information was anonymously downloaded in a monthly report, and the information on patients with more than one visit was updated. The registry includes demographic, clinical and disease characteristics, including the Montreal Classification, medical treatment, surgeries and hospitalizations for crisis. Data regarding infection with Clostridium difficile(C. difficile) were incorporated in the registry in 2014. Data for patients who received consultations as second opinions and continued treatment at this institution were also analyzed. RESULTS: The study included 716 patients with IBD: 508 patients(71%) were diagnosed with ulcerative colitis(UC), 196 patients(27%) were diagnosed with Crohn's disease(CD) and 12 patients(2%) were diagnosed with unclassifiable IBD. The UC/CD ratio was 2.6/1. The median age was 36 years(range 16-88), and 58% of the patients were female, with a median age at diagnosis of 29 years(range 5-76). In the past 15 years, a sustained increase in the number of patients diagnosed with IBD was observed, where 87% of the patients were diagnosed between the years 2001 and 2015. In the cohort examined in the present study, extensive colitis(50%) and colonic involvement(44%) predominated in the patients with UC and CD, respectively. In CD patients, nonstricturing/non-penetrating behavior was more frequent(80%), and perianal disease was observed in 28% of the patients. There were significant differences in treatment between UC and CD, with a higher use of corticosteroids, and immunosuppressive and biological therapies was observed in the patients with CD(P < 0.05 and P < 0.01). Significant surgical differences were also observed: 5% of the UC patients underwent surgery, whereas 38% of the CD patients required at least one surgery(P < 0.01). The patients with CD were hospitalized more often during their disease course than the patients with UC(55% and 35% of the patients, respectively; P < 0.01). C. difficile infection was acquired by 5% of the patients in each group at some point during the disease course. Nearly half of the patients consulted at the institution for a second opinion, and 32% of these individuals continued treatment at the institution. CONCLUSION: IBD has continued to increase in the study cohort, slowly approaching the level reported in developed countries.

The Role of PET/CT Imaging in Lung Cancer

Lung cancer presents a high prevalence around the world and constitutes the leading cause of death due to cancer in men and women. Proper staging is essential in order to provide the correct treatment and estimate survival. PET/CT imaging is the method of choice for the staging of lung cancer patients with a better performance than the traditional imaging methods. It allows defining the primary tumor more clearly, separating it from the atelectasis and neighboring fibrotic changes, thus achieving an improved T evaluation while optimizing the target definition for radiotherapy. It presents greater sensibility and specificity than Computed Tomography (CT) when assessing the extra-thoracic and thoracic lymph nodes, modifying the N stage with regard to the traditional study in up to 25% of patients. PET/CT imaging is the method with the best performance for detecting extra-encephalic metastasis and for detecting unexpected lesions in up to 30% of patients. In order to achieve proper encephalic staging, a Magnetic Resonance Imaging scan must be performed or, a CT scan of the brain concomitant to the PET/CT. One to eight percent of lung cancer patients who are subjected to PET/CT imaging present a synchronous secondary extra pulmonary or pulmonary cancer when being diagnosed, with significant prognostic and therapeutic implications. PET/CT imaging has proven to be a valuable tool for survival prognosis with regard to the initial uptake of the tumor and the early response (interim) or final response to chemotherapy, although it is not routinely used for evaluating therapy. The PET/MRI is an emerging hybrid technique with certain advantages, such as less exposure to radiation in comparison to PET/CT and greater contrast resolution of the MRI images. However, its usefulness has not been validated yet for lung cancer patients.

Minimal volume vitrification of epididymal spermatozoa results in successful in vitro fertilization and embryo development in mice

This study compared three cryopreservation protocols on sperm functions, IVF outcomes, and embryo development. Epididymal spermatozoa cryopreserved using slow-cooling （18% w/v raffinose, RS-C） were compared with spermatozoa vitrified using 0.25 M sucrose （SV） or 18% w/v raffinose （RV）. The motility, vitality, and DNA damage （TUNEL assay） of fresh control （FC） spermatozoa were compared with post-thawed or warmed RS-C, RV, and SV samples. Mouse oocytes （n = 267） were randomly assigned into three groups for insemination： RV （n = 102）, RS-C （n = 86）, and FC （n = 79）. The number and the proportion of two-cell embryos and blastocysts from each treatment were assessed. Sperm motility （P 〈 0.01） and vitality （P 〈 0.05） were significantly reduced after vitrification compared with slow-cooled spermatozoa. However, DNA fragmentation was significantly reduced in spermatozoa vitrified using sucrose （15 - 1.8% [SV] vs 26 - 2.8% [RV] and 27 - 1.2% [RS-C]; P 〈 0.01）. Although the number of two-cell embryos produced by RS-C, RV, and FC spermatozoa was not significantly different, the number of blastocysts produced from two-cell embryos using RV spermatozoa was significantly higher than FC spermatozoa （P = 0.0053）. This simple, small volume vitrification protocol and standard insemination method allows successful embryo production from small numbers of epididymal spermatozoa and may be applied clinically to circumvent the need for ICSI, which has the disadvantage of bypassing sperm selection.