Search

STUDY QUESTION Do low levels of anti-Müllerian hormone (AMH) or antral follicle count (AFC) properly predict miscarriage in young women conceiving with ART? SUMMARY ANSWER Low ovarian reserve, as indicated by AMH or AFC, is not associated with miscarriage in young women conceiving with ART. WHAT IS KNOWN ALREADY Presently, the impact of low ovarian reserve on the risk of miscarriage remains controversial. Some studies have reported an association between serum AMH levels and AFC and miscarriage, but others have failed to confirm these findings. The main limitation that undermines the reliability and consistency of the results is the confounding effect of female age. Indeed, after 35 years of age, on the one hand, the risk of miscarriage starts increasing because of impaired oocyte quality while, on the other, the physiological decline in AMH and AFC levels continues, thus hampering the possibility to properly explore the real effects of reduced ovarian reserve. Indeed, the two processes, i.e., the gradual loss of resting primordial follicles and the loss of oocyte quality, progress in parallel. In other words, the older the woman becomes, the higher is the risk of miscarriage, but one cannot distinguish between the effects of biological aging on oocyte quality and those mediated by a lower ovarian reserve. STUDY DESIGN, SIZE, DURATION The present retrospective monocentric cohort study was carried out at Fondazione IRCSS Ca Granda Ospedale Maggiore Policlinico, Milan. All women referred to the ART Unit between 2014 and 2021 and who underwent either conventional in vitro fertilization (c-IVF), intracytoplasmatic sperm injection (ICSI) or intrauterine insemination (IUI) were reviewed. Only women younger than 35 were eligible because, up to this age, the risk of miscarriage is steady and not strictly related to age. PARTICIPANTS/MATERIALS, SETTING, METHODS Women younger than 35 who achieved a singleton clinical pregnancy with c-IVF, ICSI or IUI were selected. Women with patent causes of recurrent miscarriage were excluded, as well as those undergoing pregnancy termination for fetal or medical causes. Women who did and did not have a pregnancy loss before 20 weeks’ gestation were compared. Detailed information was obtained from charts of the consulting patients. ART procedures were performed according to the standardized policy of our Unit. All women underwent serum AMH measurement and a transvaginal assessment of AFC prior to initiation of treatment. AMH levels were measured by a commercially available ELISA assay. To assess AFC, all identifiable antral follicles 2–10 mm in diameter at ultrasound were recorded. The primary outcome was the risk of miscarriage for women with serum AMH levels below 5 pmol/l. MAIN RESULTS AND THE ROLE OF CHANCE There were 538 women were included, of whom 92 (17%) had a miscarriage. The areas under the ROC curves for prediction of miscarriage based on AMH levels and AFC were 0.51 (95%CI: 0.45-0.58) and 0.52 (95%CI: 0.45-0.59), respectively. The OR of miscarriage for women with serum AMH levels below 5.0 pmol/l was 1.10 (95%CI: 0.51-2.36); the adjusted OR was 1.12 95%CI: 0.51-2.45). Analyses were repeated considering other thresholds for AMH (2.9, 3.6 and 7.9 pmol/l) and for AFC (thresholds of 7 and 10). No associations emerged. LIMITATIONS, REASONS FOR CAUTION The retrospective design of the study hampered the collection of more precise but potentially relevant clinical information of the couples. We did not exclude women suffering from PCOS, a condition possibly associated with miscarriage. Moreover, the baseline characteristics of women who did and did not have a miscarriage differed in some characteristics. Thus, we adjusted the OR using a multivariate analysis, but we cannot fully exclude residual confounding effects. Finally, our results cannot be inferred to women older than 35. The mechanisms causing premature exhaustion of ovarian reserve may be different in younger and older women and this may lead to a different impact on the risk of miscarriage. WIDER IMPLICATIONS OF THE FINDINGS Women embarking on ART with low ovarian reserve should be informed of their likely poor response to ovarian stimulation but can be reassured that, if conception occurs, their risk of miscarriage is not increased. STUDY FUNDING/COMPETING INTEREST(S) This study was partially funded by Italian Ministry of Health - Current research IRCCS. Prof. Somigliana reports grants from Ferring and honoraria for lectures from Merck-Serono and Gedeon-Richter. All the other authors do not have any competing interest to declare. TRIAL REGISTRATION NUMBER N/A WHAT DOES THIS MEAN FOR PATIENTS? The impact of low ovarian reserve on the risk of miscarriage remains controversial. Serum anti-Müllerian hormone (AMH) and ultrasound assessment of antral follicle count (AFC) are both biomarkers commonly used to assess ovarian reserve. Some studies have reported an association between AMH and/or AFC and miscarriage, but others failed to confirm these findings. In this regard, female age represents the most important limitation for the reliability and consistency of the results. Indeed, after 35 years of age, the risk of miscarriage starts increasing because of impaired oocyte quality but, at the same time, there is a physiological decline in AMH and AFC levels. In other words, the older the woman, the higher the risk of miscarriage. In ART, however, the effects of biological aging on oocyte quality and those mediated by a lower ovarian reserve cannot be distinguished during assisted reproduction technology (ART) treatments. To address this issue, we retrospectively selected young women who had a clinical pregnancy through ART and evaluated whether low levels of AMH or AFC could predict miscarriage. All women under 35 who were referred to our ART Unit and who underwent ART procedures were recruitable. This study included 538 women, of whom 92 (17%) had a miscarriage. No associations emerged in terms of AMH levels and AFC between women who did and did not have a pregnancy loss before 20 weeks’ gestation. Despite some limitations, we can conclude that serum AMH levels are not associated with miscarriage in young women conceiving with ART. Young women embarking on ART with low ovarian reserve should therefore be informed of their poorer response to ovarian stimulation but can be reassured that, if conception occurs, their risk of miscarriage is not increased.

The high-throughput quantification of intact proteoforms using a label-free approach is typically performed on proteins in the 0-30 kDa mass range extracted from whole cell or tissue lysates. Unfortunately, even when high-resolution separation of proteoforms is achieved by either high performance liquid chromatography or capillary electrophoresis, the number of proteoforms that can be identified and quantified is inevitably limited by the inherent sample complexity. Here we benchmark label-free quantification of proteoforms of Escherichia coli by applying gas-phase fractionation (GPF) via field asymmetric ion mobility spectrometry (FAIMS). Recent advances in Orbitrap instrumentation have enabled the acquisition of high-quality intact and fragmentation mass spectra without the need for averaging time-domain transients prior to Fourier transform. The resulting speed improvements allowed for the application of multiple FAIMS compensation voltages in the same liquid chromatography-tandem mass spectrometry experiment without increasing the overall data acquisition cycle. As a result, the application of FAIMS to label-free quantification based on intact mass spectra substantially increases the number of both identified and quantified proteoforms without penalizing quantification accuracy in comparison to traditional label-free experiments that do not adopt GPF.

Terra rossa is a reddish clay soil which is often present on the surface of limestone in regions with a Mediterranean-type climate. Its genesis is a controversial subject in terms of the origin of the parent material, from the residuum of underlying (carbonate/dolomite) bedrock in the absence/presence of an external silicate contribution (e.g., aeolian dust). Within this context the main goal of the present work was the understanding of the geochemical processes leading to the formation of the terra rossa starting from the carbonate bedrock. We report the results of a multi-method analysis on a terra rossa deposit occupying the bottom of a Quaternary karst depression on Mesozoic limestones exposed in the Murge area (Apulia Foreland, southern Italy). Geological, petrographic, textural, and chemical data were collected on karst products (reddish calcite incrustations and nodules, and fine-grained portion of terra rossa) by a detailed field mapping, optical microscopy, XRF and fusion ICP/MS analyses and by scanning electron microscope. New collected data show that the mineralogical composition of reddish incrustations and nodules is comparable, consisting of fibrous and impure calcite, detrital fragments of quartz, K-feldspar, zircon and authigenic minerals as (Mn, Ba, Ca) phases, (Al, Si, Mn, Fe, Mg, Ba, Ca) minerals, Fe-kaolinite and anatase. The prevailing minerals, instead, in the fine-grained portion of terra rossa are hematite, kaolinite, and goethite. Based on the chemical composition, and especially on REE patterns, a progressive interaction of silicate aqueous solutions (with Al, Si, Fe), containing pelite material, with the calcareous bedrock, as a source of carbonic acid, was the process driving the formation of terra rossa. Obtained results add new elements to the definition of the long-lasting question about the genetic processes responsible for the formation of terra rossa, corroborating their polygenetic origin, as result of limestone alteration in conjunction with the chemical interaction with allochthonous siliciclastic material.

Purpose: Recent trends in US healthcare have seen growing consolidation of healthcare providers, including radiology groups, with fewer and larger radiology groups. We assessed recent trends and characteristics in radiology group network market share (NMS) across the US among Medicare beneficiaries. Methods: Using freely available datasets CareSet DocGraph Hop Teaming, Medicare Physician Compare, and Medicare Physician and Other Supplier Public Use File (PUF), all radiologists were identified and associated to group practices annually between 2014 and 2017. Radiology groups outside the US, not present in all three databases, or with only one radiologist were excluded. The annual frequency of radiological exams performed was determined from the PUF and used to calculate the percentage of magnetic resonance and computed tomography (MR/CT) imaging performed as well as the number of beneficiaries undergoing MR/CT per radiology group. Physician referrers without evaluation and management codes for office visits in the PUF file were excluded from the DocGraph file. Provider connections were geospatially mapped and plotted. The percentage of radiology group NMS was calculated as the number of patients a group received, divided by the total number of potential and actual connections, and multiplied by 100. Univariate analysis of radiology group NMS was performed against a variety of characteristics and compared using Kruskal-Wallis and Dunn tests, as appropriate. Univariate linear regression was used to assess the association between NMS and calendar year, as well as average wait time and calendar year. Multivariate linear regression was used to model the radiology group cumulative normalized percentile NMS, with multiple predictors for the year 2017. Results: Between 2014 and 2017, 1,764 unique radiology groups were identified, representing 17,879 radiologists in 2014, 18,143 radiologists in 2015, 20,915 radiologists in 2016, and 22,187 radiologists in 2017, with an average NMS per group for 2014 of 14.8%, 2015 of 14.5%, 2016 of 13.6%, and 2017 of 13.1%, demonstrating a small but statistically significant negative trend over four years (2014-2017), with a 0.6% decrease per year (P < 0.001). Average day weight across years (2014-2017) demonstrated a slight upward trend, with a 0.3% increase per year. The yearly percentage of MR/CT studies across all groups was 24.6-26.6%, with the most performed studies being chest radiography, mammography, and CT of the head without contrast. Univariate analysis of radiology group NMS was not significantly different between academic and nonacademic groups for all years (P > 0.05) but was significantly different for radiology-only versus multispecialty groups across all years (P < 0.05). Multivariate linear regression on 2017 data demonstrated statistically significant independent negative predictors for NMS including larger group size (50-99, >=100), higher practice MR/CT imaging percentage, and South location, while the increasing log of the number of beneficiaries undergoing MR/CT was a positive predictor, with an adjusted R-squared of 0.56 (all P < 0.01). Conclusions: Among Medicare beneficiaries from 2014 to 2017, radiology group NMS (mean 14%, median 9.0%, IQR 4-19.2%), slightly decreased over time by 0.6% per year, despite occurring during a period of widespread practice consolidation. The negative predictors for NMS at the group level included larger group size, South region, increased average wait time, and higher MR/CT imaging percentage, while the positive predictor was the increased number of beneficiaries undergoing MR/CT. Of these predictors, radiology groups are most likely to increase NMS by decreasing average wait times.

Human biology is tightly linked to proteins, yet most measurements do not precisely determine alternatively spliced sequences or posttranslational modifications. Here, we present the primary structures of ~30,000 unique proteoforms, nearly 10 times more than in previous studies, expressed from 1690 human genes across 21 cell types and plasma from human blood and bone marrow. The results, compiled in the Blood Proteoform Atlas (BPA), indicate that proteoforms better describe protein-level biology and are more specific indicators of differentiation than their corresponding proteins, which are more broadly expressed across cell types. We demonstrate the potential for clinical application, by interrogating the BPA in the context of liver transplantation and identifying cell and proteoform signatures that distinguish normal graft function from acute rejection and other causes of graft dysfunction.

Blood serum and plasma are arguably the most commonly analyzed clinical samples, with dozens of proteins serving as validated biomarkers for various human diseases. Top-down proteomics may provide additional insights into disease etiopathogenesis since this approach focuses on protein forms, or proteoforms, originally circulating in blood, potentially providing access to information about relevant post-translational modifications, truncation, single amino acid substitutions and many other sources of protein variation. However, the vast majority of proteomic studies on serum and plasma are carried out using peptide-centric, bottom-up approaches which cannot recapitulate the original proteoform content of samples. Lengthy sample preparation and the need for extensive prefractionation to mitigate proteoform dynamic range issues are likely factors preventing clinical laboratories from routinely performing top-down experiments. In this study, we describe a straightforward protocol for intact proteoform sample preparation based on depletion of albumin and immunoglobulins followed by simplified fractionation of remaining serum proteins via polyacrylamide gel electrophoresis. After molecular weight-based fractionation, we supplemented the traditional liquid chromatography tandem mass spectrometry (LC-MS2) data acquisition with high-field asymmetric waveform ion mobility spectrometry (FAIMS), which served as an additional separation dimension to further simplify serum proteoforms mixtures. This LC-FAIMS-MS2 method led to the identification of over 1,000 serum proteoforms

Cachexia is a debilitating wasting syndrome and highly prevalent comorbidity in cancer patients. It manifests especially with energy and mitochondrial metabolism aberrations that promote tissue wasting. We recently identified nicotinamide adenine dinucleotide (NAD+) loss to associate with muscle mitochondrial dysfunction in cancer hosts. In this study we confirm that depletion of NAD+ and downregulation of Nrk2, an NAD+ biosynthetic enzyme, are common features of severe cachexia in different mouse models. Testing NAD+ repletion therapy in cachectic mice reveals that NAD+ precursor, vitamin B3 niacin, efficiently corrects tissue NAD+ levels, improves mitochondrial metabolism and ameliorates cancer- and chemotherapy-induced cachexia. In a clinical setting, we show that muscle NRK2 is downregulated in cancer patients. The low expression of NRK2 correlates with metabolic abnormalities underscoring the significance of NAD+ in the pathophysiology of human cancer cachexia. Overall, our results propose NAD+ metabolism as a therapy target for cachectic cancer patients.