Heavy metals in follicular fluid affect the ultrastructure of the human mature cumulus-oocyte complex

It is known that exposure to heavy metals such as lead (Pb) and cadmium (Cd) has several adverse effects, particularly on the human reproductive system. Pb and Cd have been associated with infertility in both men and women. In pregnant women, they have been associated with spontaneous abortion, preterm birth, and impairment of the development of the fetus. Since these heavy metals come from both natural and anthropogenic activities and their harmful effects have been observed even at low levels of exposure, exposure to them remains a public health issue, especially for the reproductive system. Given this, the present study aimed to investigate the potential reproductive effects of Pb and Cd levels in the follicular fluid (FF) of infertile women and non-smokers exposed to heavy metals for professional reasons or as a result of living in rural areas near landfills and waste disposal areas in order to correlate the intrafollicular presence of these metals with possible alterations in the ultrastructure of human cumulus-oocyte complexes (COCs), which are probably responsible for infertility. Blood and FF metals were measured using atomic Citation: Miglietta, S.; Cristiano, L.; Battaglione, E.; Macchiarelli, G.; Nottola, S.A.; De Marco, M.P.; Costanzi, F.; Schimberni, M.; Colacurci, N.; Caserta, D.; Familiari, G. Heavy Metals in Follicular Fluid Affect the Ultrastructure of the Human Mature Cumulus-Oocyte Complex. Cells 2023, 12, 2577. https://doi.org/10.3390/cells12212577 Academic Editor: Elena Llano Received: 11 October 2023 Revised: 30 October 2023 Accepted: 31 October 2023 Published: 5 November 2023 Copyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/license s/by/4.0/). Cells 2023, 12, 2577 2 of 24 absorption spectrometry. COCs corresponding to each FF analyzed were subjected to ultrastructural analyses using transmission electron microscopy. We demonstrated for the first time that intrafollicular levels of Pb (0.66 µg/dL–0.85 µg/dL) and Cd (0.26 µg/L–0.41 µg/L) could be associated with morphological alterations of both the oocyte and cumulus cells’ (CCs) ultrastructure. Since blood Cd levels (0.54 µg/L–1.87 µg/L) were above the current reference values established by the guidelines of the Agency for Toxic Substances and Disease Registry (ATSDR) and the Environmental Protection Agency (EPA) (0.4 µg/L), whereas blood Pb levels (1.28 µg/dL–3.98 µg/dL) were below the ATSDR reference values (≤5 µg/dL), we believe that these alterations could be due especially to Cd, even if we cannot exclude a possible additional effect of Pb. Our results highlighted that oocytes were affected in maturation and quality, whereas CCs showed scarcely active steroidogenic elements. Regressing CCs, with cytoplasmic alterations, were also numerous. According to Cd’s endocrine-disrupting activity, the poor steroidogenic activity of CCs might correlate with delayed oocyte cytoplasmic maturation. So, we conclude that levels of heavy metals in the blood and the FF might negatively affect fertilization, embryo development, and pregnancy, compromising oocyte competence in fertilization both directly and indirectly, impairing CC steroidogenic activity, and inducing CC apoptosis

Hormonal therapy followed by chemotherapy or the reverse sequence as first-line treatment of hormone-responsive, human epidermal growth factor receptor-2 negative metastatic breast cancer patients: Results of an observational study

Introduction Although hormonal-therapy is the preferred first-line treatment for hormone-responsive, HER2 negative metastatic breast cancer, no data from clinical trials support the choice between hormonal-therapy and chemotherapy. Methods Patients were divided into two groups according to the treatment: chemotherapy or hormonal-therapy. Outcomes in terms of clinical benefit and median overall survival (OS) were retrospectively evaluated in the two groups. To calculate the time spent in chemotherapy with respect to OS in the two groups, the proportion of patients in chemotherapy relative to those present in either group was computed at every day from the start of therapy. Results From 1999 to 2013, 119 patients received first-line hormonal-therapy (HT-first group) and 100 first-line chemotherapy (CT-first group). Patients in the CTfirst group were younger and with poorer prognostic factors as compared to those in HT-first group. Clinical benefit (77 vs 81%) and median OS (50.7 vs 51.1 months) were similar in the two groups. Time spent in chemotherapy was significantly longer during the first 3 years in CT-first group (54-34%) as compared to the HT-first group (11-18%). This difference decreased after the third year and overall was 28% in the CT-first group and 18% in the HT-first group. Conclusions The sequence first-line chemotherapy followed by hormonal-therapy, as compared with the opposite sequence, is associated with a longer time of OS spent in chemotherapy. However, despite the poorer prognostic factors, patients in the CTfirst group had a superimposable OS than those in the HT-first group

Effects of simulated microgravity In vitro on human metaphase II oocytes: an electron microscopy-based study

The Gravity Force to which living beings are subjected on Earth rules the functionality of most biological processes in many tissues. It has been reported that a situation of Microgravity (such as that occurring in space) causes negative effects on living beings. Astronauts returning from space shuttle missions or from the International Space Station have been diagnosed with various health problems, such as bone demineralization, muscle atrophy, cardiovascular deconditioning, and vestibular and sensory imbalance, including impaired visual acuity, altered metabolic and nutritional status, and immune system dysregulation. Microgravity has profound effects also on reproductive functions. Female astronauts, in fact, suppress their cycles during space travels, and effects at the cellular level in the early embryo development and on female gamete maturation have also been observed. The opportunities to use space flights to study the effects of gravity variations are limited because of the high costs and lack of repeatability of the experiments. For these reasons, the use of microgravity simulators for studying, at the cellular level, the effects, such as those, obtained during/after a spatial trip, are developed to confirm that these models can be used in the study of body responses under conditions different from those found in a unitary Gravity environment (1 g). In view of this, this study aimed to investigate in vitro the effects of simulated microgravity on the ultrastructural features of human metaphase II oocytes using a Random Positioning Machine (RPM). We demonstrated for the first time, by Transmission Electron Microscopy analysis, that microgravity might compromise oocyte quality by affecting not only the localization of mitochondria and cortical granules due to a possible alteration of the cytoskeleton but also the function of mitochondria and endoplasmic reticulum since in RPM oocytes we observed a switch in the morphology of smooth endoplasmic reticulum (SER) and associated mitochondria from mitochondria-SER aggregates to mitochondria–vesicle complexes. We concluded that microgravity might negatively affect oocyte quality by interfering in vitro with the normal sequence of morphodynamic events essential for acquiring and maintaining a proper competence to fertilization in human oocyte

Unusual Osteoblastic Secondary Lesion as Predominant Metastatic Disease Spread in Two Cases of Uterine Leiomyosarcoma

Uterine smooth muscle tumors range from benign leiomyomata to low- and high-grade leiomyosarcomas. A leiomyosarcoma is a rare malignant smooth muscle tumor that infrequently metastasizes to the bone. In fact, initial presentation or recurrence as osseous metastases is extremely uncommon in patients with a history of leiomyosarcoma. On imaging, these bone lesions generally appear as lytic foci. The authors report here two cases of osteoblastic bone lesions in leiomyosarcoma of the uterus with predominant metastatic lesions localized in the bone

HDL cholesterol is a strong determinant of endothelial progenitor cells in hypercholesterolemic subjects

Endothelial progenitor cells (EPC) can repair the endothelial layer and are considered a component of the cardiovascular system. EPC number and function may change under pathological conditions, including cardiovascular risk factors. The study was carried out to investigate circulating EPC number, in vitro function and relationship with LDL-C, HDL-C and endothelium-dependent vasodilatation in hypercholesterolemic subjects. Forty-one male and 39 female subjects, age>35 and130 mg/dl with normal (>= 50 mg/dl females and >= 40 mg/dl males) or low HDL-C, absence of any concomitant disorders and/or drug treatment, at their first diagnosis of hypercholesterolemia, were consecutively recruited in the Outpatient Service of the Medical Pathophysiology Department of Rome Sapienza University. In high LDL-C patients, circulating EPC number was decreased and EPC capability to migrate was impaired as well. This pattern was far less evident in the normal HDL-C subgroup. The endothelium-dependent vasodilatation (EDV) was significantly decreased according to the HDL-C decrease in male but not in female subjects. Univariate analysis showed a direct correlation between EPC number and EDV, and the association persisted after adjustment for sex, age and HDL-C, which were all significantly correlated to EDV, which may suggest a protective role of EPC on endothelium in vivo. Our study documented that, in hypercholesterolemic subjects, HDL-C is a strong determinant of EPC number and function, and EPC number decrease is an independent risk factor for endothelial dysfunction. (C) 2010 Elsevier Inc. All rights reserved

MALDI-TOF mass spectrometry for the rapid identification of aetiological agents of sepsis

Introduction: The MALDI-TOF has recently become part of the methods of microbiological investigation in many laboratories of bacteriology with advantages both practical and economical.The use of this technique for the rapid identification of the causative agents of sepsis is of strategic importance to the ability to provide the clinician with useful information for a prompt and rapid establishment of an empirical antimicrobial “targeted” therapy. Methods: It was tested a total of 343 positive blood culture bottles from 211 patients. The samples after collection were incubated in the BACTEC FX (Becton Dickinson, USA). From these bottles were taken a few milliliters of broth culture and transferred into a vacutainer tube containing gel. This was centrifuged, the supernatant was decanted, and finally recovered the bacterial suspension on the gel. With micro-organisms recovered in this way, after several washes with distilled water, was prepared a slide for microscopic examination with Gram stain, and a plate for mass spectrometry (MS-Vitek, bioMérieux, France).Then, the same samples were inoculated on solid agar media according to the protocol in use in our laboratory.The next day was checked the possible bacterial growth on solid media; we then proceeded to the identification of the colonies by Vitek MS and / or with the system Vitek2 (bioMérieux, France). Results: 258 (75.2%) positive vials show concordant results between direct identification and identification after growth on agar. For 83 (24.2%) positive bottles there has been full compliance with the microscopic examination but not with culture. In particular, two bottles (0.6%) have given complete discordance between the direct identification and that after growth. Conclusions: The protocol we use for the direct identification of organisms responsible for sepsis, directly on positive bottles, seems to be a quick and inexpensive procedure, which in less than 60 minutes can give valuable feedback to the clinician

Prevalence of Clostridium difficile infections in Prato hospital in the years 2010-2011

Clostridium difficile, a Gram positive, spore-forming, anaerobic bacillus, is an important nosocomial enteric pathogen causing diarrhoea and pseudomembranous colitis. Aim of this study was to evaluate the prevalence of Clostridium difficile in Prato Hospital. Stool samples were collected from 1197 patients hospitalized from January 2010 to December 2011. In all the samples the common antigen GDH was investigated and only in samples positive for the antigen the presence of the A and B toxins was investigated. Our results showed that 170/1197 samples (14%) were positive for the antigen, and of these 170 patients, 84 (49%) were found positive also for the toxins. In addition the percentage of samples positive for toxins was higher in 2010 (8.6%) than in 2011 (5.9%)

Detection of T. vaginalis,M. hominis,M. genitalium, C. trachomatis, N. gonorrhoeae and U. urealyticum using Multiplex PCR

Intoduction. The sexually transmitted diseases include a large group of infections affecting both the sexes. In this study we evaluated the prevalence of Trichomonas vaginalis, Mycoplasma hominis, Mycoplasma genitalium, Chlamydia trachomatis, Neisseria gonorrhoeae and Ureaplasma urealyticum in the Prato area during the period September 2010 – July 2011. Methods.We analysed different kind of samples (urine, endocervical swabs, urethral swabs, seminal fluids) from hospitalized patients or referred to the Prato clinic subjects.The DNA was obtained using EZ1-DNA extraction kit and EZ1 instrument.The DNA was then amplified using the Seeplex STD6 kit (Seegene, Korea), identifying multiple pathogens simultaneously (T. vaginalis, M. hominis, M. genitalium, C. trachomatis, N. gonorrhoeae e U. urealyticum). The revelation was performed by electrophoresis on microchip (instrument Multina, Shimadzu, Japan). Results. 1136 samples from Italian and foreign patients were examined: 876 were endocervical swabs (77%), 103 urethral swabs (9%), 103 seminal fluids (9%), and 54 urines (5%). The number of females was higher than males [894 (78.7%) vs 242 (21.3%)]; the mean age of females was 37.0±11.6 years, whereas that of males was 41.5 ±12.63 years.The prevalence of urogenital pathogens was: 15 positive samples for T. vaginalis (1.3%), 56 for M. hominis (4.9%), 13 for M. genitalium (1.1%), 28 for C. trachomatis (2.5%), 8 for N. gonorrhoeae (0.7%) and 87 for U. urealyticum (7.7%).Among all positive, 25 subjects were positive for more than one pathogen and in particular: one was positive for the presence of 4 pathogens, five presented 3 pathogens simultaneously and the remaining nineteen for 2 pathogens. Conclusions. This study provides data on the prevalence of sexually transmitted diseases in the hospital of Prato

Screening for Klebsiella pneumoniae producing carbapenemase in Prato, Italy

In this study, the prevalence of carbapenemase in the area of Prato (Italy), from January 2011 to August 2013, is reported. Samples showing carbapenem resistance were tested for lactamase production with disc diffusion and molecular methods. Totally, 48 out of 1542 patients with K. pneumoniae showed carbapenem resistance (42 KPC 5 MBL and 1 OXA-48).</p