RESUMO
BACKGROUND: One of the causes of male infertility is associated with altered spermatozoa motility. These sperm features are frequently analyzed by image-based approaches, which, despite allowing the acquisition of crucial parameters to assess sperm motility, they are unable to provide details regarding the flagellar beating forces, which have been neglected until now. RESULTS: In this work we exploit Fluidic Force Microscopy to investigate and quantify the forces associated with the flagellar beating frequencies of human spermatozoa. The analysis is performed on two groups divided according to the progressive motility of semen samples, as identified by standard clinical protocols. In the first group, 100% of the spermatozoa swim linearly (100% progressive motility), while, in the other, spermatozoa show both linear and circular motility (identified as 80 - 20% progressive motility). Significant differences in flagellar beating forces between spermatozoa from semen sample with different progressive motility are observed. Particularly, linear motile spermatozoa exhibit forces higher than those with a circular movement. CONCLUSIONS: This research can increase our understanding of sperm motility and the role of mechanics in fertilization, which could help us unveil some of the causes of idiopathic male infertility.
Assuntos
Infertilidade Masculina , Sêmen , Humanos , Masculino , Motilidade dos Espermatozoides , Análise do Sêmen , EspermatozoidesRESUMO
BACKGROUND: Endometriosis is a gynecological disease characterized by the presence of endometrial tissue in abnormal locations, leading to severe symptoms, inflammation, pain, organ dysfunction, and infertility. Surgical removal of endometriosis lesions is crucial for improving pain and fertility outcomes, with the goal of complete lesion removal. This study aimed to analyze the location and expression patterns of poly (ADP-ribose) polymerase 1 (PARP-1), epithelial cell adhesion molecule (EpCAM), and folate receptor alpha (FRα) in endometriosis lesions and evaluate their potential for targeted imaging. METHODS: Gene expression analysis was performed using the Turku endometriosis database (EndometDB). By immunohistochemistry, we investigated the presence and distribution of PARP-1, EpCAM, and FRα in endometriosis foci and adjacent tissue. We also applied an ad hoc platform for the analysis of images to perform a quantitative immunolocalization analysis. Double immunofluorescence analysis was carried out for PARP-1 and EpCAM, as well as for PARP-1 and FRα, to explore the expression of these combined markers within endometriosis foci and their potential simultaneous utilization in surgical treatment. RESULTS: Gene expression analysis revealed that PARP-1, EpCAM, and FOLR1 (FRα gene) are more highly expressed in endometriotic lesions than in the peritoneum, which served as the control tissue. The results of the immunohistochemical study revealed a significant increase in the expression levels of all three biomarkers inside the endometriosis foci compared to the adjacent tissues. Additionally, the double immunofluorescence analysis consistently demonstrated the presence of PARP-1 in the nucleus and the expression of EpCAM and FRα in the cell membrane and cytoplasm. CONCLUSION: Overall, these three markers demonstrate significant potential for effective imaging of endometriosis. In particular, the results emphasize the importance of PARP-1 expression as a possible indicator for distinguishing endometriotic lesions from adjacent tissue. PARP-1, as a potential biomarker for endometriosis, offers promising avenues for further investigation in terms of both pathophysiology and diagnostic-therapeutic approaches.
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Endometriose , Molécula de Adesão da Célula Epitelial , Receptor 1 de Folato , Poli(ADP-Ribose) Polimerase-1 , Endometriose/metabolismo , Endometriose/cirurgia , Endometriose/genética , Endometriose/diagnóstico , Endometriose/patologia , Feminino , Humanos , Poli(ADP-Ribose) Polimerase-1/metabolismo , Poli(ADP-Ribose) Polimerase-1/genética , Receptor 1 de Folato/genética , Receptor 1 de Folato/metabolismo , Molécula de Adesão da Célula Epitelial/genética , Molécula de Adesão da Célula Epitelial/metabolismo , Adulto , Biomarcadores/metabolismo , Imuno-Histoquímica , Endométrio/metabolismo , Endométrio/patologia , Endométrio/cirurgiaRESUMO
BACKGROUND AND AIMS: Obesity is the most common health issue in women of reproductive age, which profoundly affects maternal-fetal health. Despite progress in understanding key inflammatory and metabolic changes, the pathogenesis of the cardiovascular phenotype of obese pregnant women remains to be fully understood. This study aimed at: (i) evaluating the changes of the renin-angiotensin system (RAS) throughout pregnancy in obese vs normal weight (control) women, and (ii) evaluating the presence of any associations between maternal hemodynamic status and RAS changes. METHODS AND RESULTS: Thirty-eight normal weight and nineteen obese pregnant women were included. Clinical assessment, blood samples and maternal hemodynamic evaluation were performed at 12, 20, 30, and 36 weeks, while ultrasound assessment was scheduled at 20, 30, and 36 weeks of gestation. Measurements of sFlt-1, PlGF, Angiotensinogen, Renin, AngII, Ang1-7, ACE and ACE2 were performed by ELISA. Our data show that normotensive obese women had lower placental blood supply, as assessed by UV-Q and UV-Q/EFW, as compared to controls, and significantly higher levels of AngII and AngII/Ang1-7 ratio, which were inversely related to placental blood supply. CONCLUSIONS: Our study shows for the first time that normotensive obese women exhibited a significant progressive increase of AngII and AngII/Ang1-7 throughout pregnancy, which were inversely related to placental blood supply as assessed by UV-Q and UV-Q/EFW. Our data shed light on the early changes in pregnant obese women and suggest that RAS dysregulation is a prerequisite rather than a consequence of hypertensive disorders of pregnancy and other maternal neonatal complications.
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Angiotensinogênio , Obesidade Materna , Sistema Renina-Angiotensina , Renina , Feminino , Humanos , Recém-Nascido , Gravidez , Ensaio de Imunoadsorção Enzimática , Estudos Longitudinais , Placenta , Obesidade Materna/sangue , Angiotensinogênio/sangue , Renina/sangueRESUMO
BACKGROUND: The primary aim of this study was to compare the quality of life between women with obstetric anal sphincter injury (OASI) and women with intact perineum or minor vaginal tears following their first vaginal birth through a validated urogynaecological questionnaire. As a secondary aim, we wanted to identify the specific symptoms for pelvic floor dysfunction after a vaginal birth. METHODS: One hundred thirty-three cases (III- and IV-degree vaginal tears) and 133 controls (intact perineum or I- and II-degree vaginal tear) were asked to fill the PFDI-20 condition-specific and quality of life survey at three and 12 months after vaginal delivery. The survey evaluates pelvic floor dysfunction symptoms through three subsections: the Pelvic Organ Prolapse Distress Inventory (POPDI), the Colorectal-Anal Distress Inventory (CRADI), and Urinary Distress Inventory, (UDI). The scoring system ranges from 0 (no distress) to 100 (maximum distress) for each subsection, subsequently summed up to obtain the summary score (0 to 300). The patients recruited were asked to complete the survey at 3- and 12-months follow-up visit. Accordingly, data collection started. Categorical variables were subjected to Chi-square test or Fisher's Exact test. Quantitative variables were compared through Student's t-test or Mann-Whitney test. RESULTS: All surveys have shown statistically significant differences when comparing the cases to the control group. Consequently, PFDI-20 has shown a strong correlation between III- and IV-grade lacerations and pelvic floor dysfunction persistence at 12 months after delivery. Intestinal symptoms were the most reported disturbances among women with previous OASI. CONCLUSIONS: Major vaginal tears have demonstrated to have a strong impact on women's quality of life up to a follow-up of 12 months. The use of PFDI-20 questionnaire is a useful and valid tool in the diagnosis and follow-up of genital prolapse, fecal and urinary incontinence in primiparous women with a history of OASI. Thus, its application in clinical practice can help offering the most adequate rehabilitative treatment.
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Incontinência Fecal , Lacerações , Diafragma da Pelve , Doenças Vaginais , Feminino , Humanos , Gravidez , Estudos de Casos e Controles , Incontinência Fecal/etiologia , Diafragma da Pelve/patologia , Qualidade de Vida , Inquéritos e QuestionáriosRESUMO
OBJECTIVE: Demonstrate the usefulness of using indocyanine green after laparoscopic ovarian detorsion to save the ovary. DESIGN: A step-by-step video demonstration of a surgical technique. SETTING: Ovarian torsion is one of the most common gynecological emergencies, mainly affecting patients younger than 20 years of age [1], and causes 2% to 7% of acute abdomens [2]. It is not advisable to routinely perform ovariectomy even with a necrotic ovary appearance [1]. Furthermore only in a small percentage of cases (16%) necrosis has been confirmed histologically [2]. Some studies have demonstrated that using indocyanine green to evaluate ovarian perfusion is inexpensive, low risk, and easily reproducible [3-5]. INTERVENTIONS: A 17-year-old patient was referred to our hospital for acute abdominal pain. Ultrasound revealed ovarian torsion; therefore, the patient underwent surgical treatment. During laparoscopy, the presence of a right ovarian torsion was confirmed. A lesion compatible with a hemorrhagic corpus luteum of 6 cm was present on the ovary affected. Before ovarian detorsion, indocyanine green was administered intravenously at a 0.5 mg/kg dose. The first aspect noted was the total lack of ovarian vascularization; then ovarian detorsion was performed. At this point, using technology of Rubina (KARL STORZ SE & Co. KG, Tuttlingen, Germany), it was possible to highlight the progressive ovarian revascularization. Ovarian reperfusion occurred starting from the ovarian hilum and ending at the periphery. We proceeded with enucleation of the hemorrhagic corpus luteum by stripping technique, with subsequent ovarian reconstruction with continuous 2-0 monofilament suture. Finally, we fixed the ovary to the stump of the right round ligament. The final view highlights good ovarian vascularization. No complications occurred; the patient was discharged on the first postoperative day. A 6-month follow-up ultrasound confirmed the recovery of the vascularization of ovary. CONCLUSION: Using indocyanine green represents a valid option to evaluate ovarian perfusion after detorsion. It could help the surgeon decide to save the ovary and thus allow fertility-sparing surgery in more cases.
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Verde de Indocianina , Laparoscopia , Feminino , Humanos , Adolescente , Torção Ovariana/cirurgia , Perfusão , Laparoscopia/métodos , Anormalidade Torcional/cirurgiaRESUMO
Respiratory infections are common in pregnancy with conflicting evidence supporting their association with neonatal congenital anomalies, especially during the first trimester. We profiled cytokine and chemokine systemic responses in 242 pregnant women and their newborns after SARS-CoV-2 infection, acquired in different trimesters. Also, we tested transplacental IgG passage and maternal vaginal-rectal microbiomes. IgG transplacental passage was evident, especially with infection acquired in the first trimester. G-CSF concentration-involved in immune cell recruitment-decreased in infected women compared to uninfected ones: a beneficial event for the reduction of inflammation but detrimental to ability to fight infections at birth. The later the infection was acquired, the higher the systemic concentration of IL-8, IP-10, and MCP-1, associated with COVID-19 disease severity. All infected women showed dysbiosis of vaginal and rectal microbiomes, compared to uninfected ones. Two newborns tested positive for SARS-CoV-2 within the first 48 h of life. Notably, their mothers had acute infection at delivery. Although respiratory infections in pregnancy are reported to affect babies' health, with SARS-CoV-2 acquired early during gestation this risk seems low because of the maternal immune response. The observed vaginal and rectal dysbiosis could be relevant for neonatal microbiome establishment, although in our series immediate neonatal outcomes were reassuring.
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COVID-19 , Disbiose , Complicações Infecciosas na Gravidez , SARS-CoV-2 , Vagina , Humanos , Feminino , Gravidez , COVID-19/imunologia , Disbiose/imunologia , Disbiose/microbiologia , Adulto , SARS-CoV-2/imunologia , Complicações Infecciosas na Gravidez/imunologia , Complicações Infecciosas na Gravidez/microbiologia , Complicações Infecciosas na Gravidez/virologia , Vagina/microbiologia , Vagina/imunologia , Vagina/virologia , Recém-Nascido , Citocinas/metabolismo , Trimestres da Gravidez/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Microbiota/imunologiaRESUMO
Among gynecological cancers, endometrial cancer is the most common in developed countries. Extracellular vesicles (EVs) are cell-derived membrane-surrounded vesicles that contain proteins involved in immune response and apoptosis. A deep proteomic approach can help to identify dysregulated extracellular matrix (ECM) proteins in EVs correlated to key pathways for tumor development. In this study, we used a proteomics approach correlating the two acquisitions-data-dependent acquisition (DDA) and data-independent acquisition (DIA)-on EVs from the conditioned medium of four cell lines identifying 428 ECM proteins. After protein quantification and statistical analysis, we found significant changes in the abundance (p < 0.05) of 67 proteins. Our bioinformatic analysis identified 26 pathways associated with the ECM. Western blotting analysis on 13 patients with type 1 and type 2 EC and 13 endometrial samples confirmed an altered abundance of MMP2. Our proteomics analysis identified the dysregulated ECM proteins involved in cancer growth. Our data can open the path to other studies for understanding the interaction among cancer cells and the rearrangement of the ECM.
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Neoplasias do Endométrio , Proteínas da Matriz Extracelular , Matriz Extracelular , Vesículas Extracelulares , Proteômica , Humanos , Feminino , Neoplasias do Endométrio/metabolismo , Neoplasias do Endométrio/patologia , Proteômica/métodos , Vesículas Extracelulares/metabolismo , Matriz Extracelular/metabolismo , Linhagem Celular Tumoral , Proteínas da Matriz Extracelular/metabolismo , Pessoa de Meia-Idade , Biologia Computacional/métodos , Metaloproteinase 2 da Matriz/metabolismoRESUMO
OBJECTIVES: To evaluate the clinical significance of nuchal translucency (NT) between the 95th-99th percentile in terms of typical and atypical chromosomal abnormalities (ACAs), associated fetal congenital defects and postnatal outcome. METHODS: A retrospective cohort study of fetuses with NT between the 95th-99th percentile. Data regarding the rate of associated fetal defects, genetic abnormalities and postnatal outcome were collected. RESULTS: A total of 306 cases of fetuses with an NT between the 95th-99th percentiles were included. The overall rate of genetic abnormalities was 12.1% (37/306). Chromosomal abnormalities were found in 10.1% (31/306) of cases and 2% were ACA (6/306). Within this group, two were pathogenic Copy Number Variants (CNVs) and four were single gene disorders. The overall rate of fetal congenital defects was 13.7% (42/306). All ACAs were found in fetuses with congenital defects. Postnatally, a new diagnosis of a single gene disorder was made in 0.85% of cases (2/236). CONCLUSIONS: The presence of an NT between the 95th-99th percentiles carries a 10-fold increased risk of fetal defects, representing an indication for referral for a detailed fetal anatomy evaluation. The risk of ACA is mainly related to the presence of fetal defects, irrespective of the combined test risk.
Assuntos
Aberrações Cromossômicas , Medição da Translucência Nucal , Gravidez , Feminino , Humanos , Primeiro Trimestre da Gravidez , Estudos Retrospectivos , Feto/diagnóstico por imagemRESUMO
Surgical resection is the cornerstone of solid tumour treatment. Current techniques for evaluating margin statuses, such as frozen section, imprint cytology, and intraoperative ultrasound, are helpful. However, an intraoperative assessment of tumour margins that is accurate and safe is clinically necessary. Positive surgical margins (PSM) have a well-documented negative effect on treatment outcomes and survival. As a result, surgical tumour imaging methods are now a practical method for reducing PSM rates and improving the efficiency of debulking surgery. Because of their unique characteristics, nanoparticles can function as contrast agents in image-guided surgery. While most image-guided surgical applications utilizing nanotechnology are now in the preclinical stage, some are beginning to reach the clinical phase. Here, we list the various imaging techniques used in image-guided surgery, such as optical imaging, ultrasound, computed tomography, magnetic resonance imaging, nuclear medicine imaging, and the most current developments in the potential of nanotechnology to detect surgical malignancies. In the coming years, we will see the evolution of nanoparticles tailored to specific tumour types and the introduction of surgical equipment to improve resection accuracy. Although the promise of nanotechnology for producing exogenous molecular contrast agents has been clearly demonstrated, much work remains to be done to put it into practice.
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Neoplasias , Cirurgia Assistida por Computador , Humanos , Meios de Contraste , Neoplasias/diagnóstico por imagem , Neoplasias/cirurgia , Cirurgia Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Imagem ÓpticaRESUMO
The in utero microbiome hypothesis has been long debated. This hypothesis will change our comprehension of the pioneer human microbiome if proved correct. In 60 uncomplicated pregnancies, we profiled the microbiome of chorionic villi (CV) and amniotic fluids (AF) in relation to maternal saliva, rectum, and vagina and the soluble cytokines cascade in the vagina, CV and AF. In our series, 12/37 (32%) AF and 10/23 (44%) CV tested positive for bacterial DNA. CV and AF harbored bacterial DNA of Streptococcus and Lactobacillus, overlapping that of the matched oral and vaginal niches, which showed a dysbiotic microbiome. In these pregnant women, the immune profiling revealed an immune hyporesponsiveness in the vagina and a high intraamniotic concentration of inflammatory cytokines. To understand the eventual role of bacterial colonization of the CV and AF and the associated immune response in the pregnancy outcome, further appropriate studies are needed. In this context, further studies should highlight if the hematogenous route could justify the spread of bacterial DNA from the oral microbiome to the placenta and if vaginal dysbiosis could favor the likelihood of identifying CV and AF positive for bacterial DNA.
Assuntos
Líquido Amniótico , Microbioma Gastrointestinal , Gravidez , Feminino , Humanos , Vilosidades Coriônicas , DNA Bacteriano/genética , Impressões Digitais de DNA , Bactérias/genética , Vagina/microbiologia , Citocinas/genéticaRESUMO
Endometrial cancer (EC) is the most common gynecologic malignancy of the endometrium. This study focuses on EC and normal endometrium phosphoproteome to identify differentially phosphorylated proteins involved in tumorigenic signalling pathways which induce cancer growth. We obtained tissue samples from 8 types I EC at tumour stage 1 and 8 normal endometria. We analyzed the phosphoproteome by two-dimensional differential gel electrophoresis (2D-DIGE), combined with immobilized metal affinity chromatography (IMAC) and mass spectrometry for protein and phosphopeptide identification. Quantities of 34 phosphoproteins enriched by the IMAC approach were significantly different in the EC compared to the endometrium. Validation using Western blotting analysis on 13 patients with type I EC at tumour stage 1 and 13 endometria samples confirmed the altered abundance of HBB, CKB, LDHB, and HSPB1. Three EC samples were used for in-depth identification of phosphoproteins by LC-MS/MS analysis. Bioinformatic analysis revealed several tumorigenic signalling pathways. Our study highlights the involvement of the phosphoproteome in EC tumour growth. Further studies are needed to understand the role of phosphorylation in EC. Our data shed light on mechanisms that still need to be ascertained but could open the path to a new class of drugs that could hinder EC growth.
Assuntos
Neoplasias do Endométrio , Fosfoproteínas , Humanos , Feminino , Fosfoproteínas/metabolismo , Cromatografia Líquida , Espectrometria de Massas em Tandem/métodos , Cromatografia de Afinidade/métodos , ProteomaRESUMO
In this study, we revealed a peculiar morphological feature of 50B11 nociceptive sensory neurons in in vitro culture related to the forskolin-induced differentiation of these cells growing upside-down on cover glass supports. Multi-photon non-linear microscopy was applied to monitor increased neurite arborization and elongation. Under live and unstained conditions, second harmonic generation (SHG) microscopy could monitor microtubule organization inside the cells while also correlating with the detection of cellular multi-photon autofluorescence, probably derived from mitochondria metabolites. Although the differentiated cells of each compartment did not differ significantly in tubulin or multi-photon autofluorescence contents, the upturned neurons were more elongated, presenting a higher length/width cellular ratio and longer neurites, indicative of differentiated cells. SHG originating from the axons' microtubules represented a proper tool to study neurons' inverted culture in live conditions without exogenous staining. This work represents the first instance of examining neuronal cell lines growing and differentiated in an upside-down orientation, allowing a possible improvement of 50B11 as a model in physiology studies of sensory neurons in peripheric nervous system disease (e.g., Fabry disease, Friedreich ataxia, Charcot-Marie-Tooth, porphyria, type 1 diabetes, Guillain-Barré syndrome in children) and analgesic drug screening.
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Axônios , Microscopia , Criança , Humanos , Colforsina/farmacologia , Axônios/fisiologia , Neuritos/fisiologia , Células Receptoras Sensoriais , Microtúbulos , Diferenciação CelularRESUMO
An improved understanding of an ovary's structures is highly desirable to support advances in folliculogenesis knowledge and reproductive medicine, with particular attention to fertility preservation options for prepubertal girls with malignant tumors. Although currently the golden standard for structural analysis is provided by combining histological sections, staining, and visible 2D microscopic inspection, synchrotron radiation phase-contrast microtomography is becoming a new challenge for three-dimensional studies at micrometric resolution. To this aim, the proper use of contrast agents can improve the visualization of internal structures in ovary tissues, which normally present a low radiopacity. In this study, we report a comparison of four staining protocols, based on iodine or tungsten containing agents, applied to bovine ovarian tissues fixed in Bouin's solution. The microtomography (microCT) analyses at two synchrotron facilities under different set-ups were performed at different energies in order to maximize the image contrast. While tungsten-based agents allow large structures to be well identified, Iodine ones better highlight smaller features, especially when acquired above the K-edge energy of the specific metal. Further scans performed at lower energy where the setup was optimized for overall quality and sensitivity from phase-contrast still provided highly resolved visualization of follicular and intrafollicular structures at different maturation stages, independent of the staining protocol. The analyses were complemented by X-ray Fluorescence mapping on 2D sections, showing that the tungsten-based agent has a higher penetration in this type of tissues.
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Imageamento Tridimensional , Iodo , Humanos , Feminino , Animais , Bovinos , Imageamento Tridimensional/métodos , Microscopia , Raios X , Microtomografia por Raio-X/métodos , Ovário , Tungstênio , Meios de Contraste/químicaRESUMO
Although only 0.8-1% of SARS-CoV-2 infections are in the 0-9 age-group, pneumonia is still the leading cause of infant mortality globally. Antibodies specifically directed against SARS-CoV-2 spike protein (S) are produced during severe COVID-19 manifestations. Following vaccination, specific antibodies are also detected in the milk of breastfeeding mothers. Since antibody binding to viral antigens can trigger activation of the complement classical - pathway, we investigated antibody-dependent complement activation by anti-S immunoglobulins (Igs) present in breast milk following SARS-CoV-2 vaccination. This was in view of the fact that complement could play a fundamentally protective role against SARS-CoV-2 infection in newborns. Thus, 22 vaccinated, lactating healthcare and school workers were enrolled, and a sample of serum and milk was collected from each woman. We first tested for the presence of anti-S IgG and IgA in serum and milk of breastfeeding women by ELISA. We then measured the concentration of the first subcomponents of the three complement pathways (i.e., C1q, MBL, and C3) and the ability of anti-S Igs detected in milk to activate the complement in vitro. The current study demonstrated that vaccinated mothers have anti-S IgG in serum as well as in breast milk, which is capable of activating complement and may confer a protective benefit to breastfed newborns.
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COVID-19 , SARS-CoV-2 , Recém-Nascido , Lactente , Feminino , Humanos , Vacinas contra COVID-19 , Lactação , Leite Humano , Proteínas do Sistema Complemento , Imunoglobulina G , Anticorpos AntiviraisRESUMO
Endometrial cancer (EC) is the most frequent gynecologic cancer in postmenopausal women. Pathogenetic mechanisms that are related to the onset and progression of the disease are largely still unknown. A multi-omics strategy can help identify altered pathways that could be targeted for improving therapeutical approaches. In this study we used a multi-omics approach on four EC cell lines for the identification of common dysregulated pathways in type 1 and 2 ECs. We analyzed proteomics and metabolomics of AN3CA, HEC1A, KLE and ISHIKAWA cell lines by mass spectrometry. The bioinformatic analysis identified 22 common pathways that are in common with both types of EC. In addition, we identified five proteins and 13 metabolites common to both types of EC. Western blotting analysis on 10 patients with type 1 and type 2 EC and 10 endometria samples confirmed the altered abundance of NPEPPS. Our multi-omics analysis identified dysregulated proteins and metabolites involved in EC tumor growth. Further studies are needed to understand the role of these molecules in EC. Our data can shed light on common pathways to better understand the mechanisms involved in the development and growth of EC, especially for the development of new therapies.
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Neoplasias do Endométrio , Multiômica , Humanos , Feminino , Neoplasias do Endométrio/metabolismo , Metabolômica , Biologia ComputacionalRESUMO
BACKGROUND: Although X-ray fluorescence microscopy is becoming a widely used technique for single-cell analysis, sample preparation for this microscopy remains one of the main challenges in obtaining optimal conditions for the measurements in the X-ray regime. The information available to researchers on sample treatment is inadequate and unclear, sometimes leading to wasted time and jeopardizing the experiment's success. Many cell fixation methods have been described, but none of them have been systematically tested and declared the most suitable for synchrotron X-ray microscopy. METHODS: The HEC-1-A endometrial cells, human spermatozoa, and human embryonic kidney (HEK-293) cells were fixed with organic solvents and cross-linking methods: 70% ethanol, 3.7%, and 2% paraformaldehyde; in addition, HEK-293 cells were subjected to methanol/ C3H6O treatment and cryofixation. Fixation methods were compared by coupling low-energy X-ray fluorescence with scanning transmission X-ray microscopy and atomic force microscopy. RESULTS: Organic solvents lead to greater dehydration of cells, which has the most significant effect on the distribution and depletion of diffusion elements. Paraformaldehyde provides robust and reproducible data. Finally, the cryofixed cells provide the best morphology and element content results. CONCLUSION: Although cryofixation seems to be the most appropriate method as it allows for keeping cells closer to physiological conditions, it has some technical limitations. Paraformaldehyde, when used at the average concentration of 3.7%, is also an excellent alternative for X-ray microscopy.
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Raios X , Humanos , Células HEK293 , Radiografia , Microscopia de Força AtômicaRESUMO
This review examined whether the absence of a genetic link with one or both parents in families using reproductive donation induced a different quality of parenting from that found in families with spontaneous conception or autologous assisted reproductive technology (AUT-ART), where the genetic mother carries the pregnancy and both parents have a genetic link with their children. MEDLINE, PsycINFO and PubMed were searched for English-language studies published from January 1993 to October 2021. A total of 45 studies were included in the systematic review, and 11 in the meta-analysis. The meta-analysis showed that in reproductive donation families, where there was no genetic link between parents and children, there were higher positive parental values (Pâ¯=â¯0.007) and lower negative parental values (Pâ¯=â¯0.007) than for parents and children in families that had spontaneously conceived. No statistically significant differences emerged when the reproductive donation families were compared with the AUT-ART families. The study showed that the quality of parenting was not conditioned by the presence or absence of a genetic link; instead, it was influenced by the processes underlying family building, such as the desire to have a child, the involvement of both parents in the childcare and the quality of disclosure.
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Relações Pais-Filho , Poder Familiar , Criança , Feminino , Humanos , Gravidez , Revelação , Doação de Oócitos , Pais , Técnicas de Reprodução AssistidaRESUMO
BACKGROUND: Intrauterine devices (IUDs) are commonly used as contraceptives worldwide. However, pregnancies in patients carrying this kind of device may occur. IUD removal when the woman wishes to continue their pregnancy may be very challenging. Only 9 manuscripts in literature reported such similar procedure. CASE PRESENTATION: We report the case of an hysteroscopic removal of IUD in a young woman at 6 weeks of gestation. DISCUSSION: The case reported highlights safety and efficacy of operative hysteroscopy as a method of IUD removal in early pregnancy, although other different methods have been reported in literature. In our opinion, maintaining a low infusion pressure during the procedure may help avoiding potential gestational sac damage and IUD displacement for better grasping.
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Dispositivos Intrauterinos , Gravidez , Feminino , Humanos , Histeroscopia/métodosRESUMO
OBJECTIVE: Many therapies have been proposed for cervical pregnancy (CP) treatment; however, there is no consensus on the best practice to adopt, mainly owing to the rarity of this condition and the lack of randomized controlled trials. Therefore, there are no clinical practice guidelines for the management of this patient set. We presented an English literature review about the hysteroscopic management of CP. DATA SOURCES: The literature review was performed according to the Preferred Reporting Items for Scoping Reviews. The search strategy aimed at identifying cases from the first patients tracked down to those diagnosed in May of 2021. We searched in PubMed, Scopus, Google Scholar, and MEDLINE databases. Mesh terms used included "Cervical Pregnancy," "Hysteroscopy," "Ectopic pregnancy," and "Resectoscopy." METHOD OF STUDY SELECTION: Case reports of randomized controlled trials, prospective controlled studies, prospective cohort studies, retrospective studies, case series, and case reports were considered eligible. Review, Letters to the Editor, and abstracts accepted at conferences were ruled out. TABULATION, INTEGRATION, AND RESULTS: We found a total of 3572 articles in all analyzed databases. A total of 2480 articles viewed were duplicated and therefore ruled out. After screening and excluding nonpertinent articles, 109 were assessed for eligibility, and 19 were included in the analysis. All articles were single case reports, small case series with no criteria selection, randomization, or study planning. We classified them as follows: cases treated with 10 mm resectoscope, with or without pretreatments of previous CP hysteroscopic approach, and cases resolved with 5 mm hysteroscopy, with or without pretreatments of previous CP hysteroscopic approach. CONCLUSION: The hysteroscopic method represents a feasible and safe approach to the CP treatment, although there are still some aspects to be clarified, such as the pretreatment need and the instruments' type and sizes based on the beta-subunit of human chorionic gonadotropin, pregnancy age, and dimension.
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Gravidez Ectópica , Feminino , Humanos , Histeroscopia/métodos , Gravidez , Gravidez Ectópica/terapia , Estudos Prospectivos , Estudos RetrospectivosRESUMO
PURPOSE: The risk of monozygotic twins (MZTs) is increased in couples undergoing assisted reproductive technology (ART) treatments. Several systematic reviews have investigated the possible determinants linked to ART, but results obtained have not been conclusive. The study aims to investigate whether the incidence of MZT differed among ART centers. METHODS: This is a multicenter retrospective cohort study using the Italian ART National Registry database and involving the centers reporting data from individual ART cycles from 2015 to 2019. To investigate the incidence of MZT, only single embryo transfer cycles were considered. Women who had sex-discordant deliveries were excluded. MZT rate was calculated as the number of multiple pregnancies (more than one gestational sac at first ultrasound) out of the total number of clinical pregnancies. A binomial distribution model was used to determine the 95% CI of the frequency of MZT. RESULTS: Eighteen centers were included, and they provided data on 10,433 pregnancies. The total number of MZT was 162, corresponding to an incidence of 1.5% (95% CI: 1.3-1.8%). The rate of MZT among centers varied between 0% (95% CI: 0.0-25.9%) and 3.2% (95% CI: 1.3-8.1%). All the 95% CIs included 1.5%, rejecting the hypothesis that the MZT rate may significantly differ among centers. CONCLUSIONS: The rate of MZT did not significantly vary among ART centers. Local factors are unlikely to explain the increased rate of MZT in ART pregnancies.