Influence of Hashimoto thyroiditis on the development of thyroid nodules and cancer in children and adolescents

It is unclear whether patients with Hashimoto thyroiditis (HT) are predisposed to develop thyroid nodules and/or thyroid cancer. The objective of our study was therefore to assess the prevalence of thyroid nodules and/or cancer in patients with HT and to look for possible prognostic factors. A retrospective survey of 904 children/adolescents with HT (709 females, 195 males) regularly followed in nine Italian centers of pediatric endocrinology was performed. Median period of follow-up was 4.5 years (1.2 to 12.8 years). We evaluated free T4, TSH, thyroid peroxidase antibody (TPOAb), thyroglobulin antibodies, and thyroid ultrasound yearly. One hundred seventy-four nodules were detected, with an annual incidence rate of 3.5%. Ten nodules were malignant (8 papillary and 2 papillary follicular variant), giving a 5.7% prevalence of cancer among patients with nodules. The severity of hypo-echogenity at ultrasound, TPOAb, and free T4 serum concentrations were predictive for the appearance of new nodules. Furthermore, a positive correlation was observed between TPOAb titer and the development of thyroid cancer. In conclusion, HT seems to influence the development of thyroid nodules, but not cancer in children and adolescents

Final height in Italian patients with congenital hypothyroidism detected by neonatal screening: A 20-year observational study

Background: Linear growth and final height are reported as normal in congenital hypothyroid patients in the neonatal screening era. Methods: We evaluated the final height in 215 patients with congenital hypothyroidism to assess if it improved over the last 2 decades. Results: Final height (-0.1∈±∈1.0 SDS) was higher than target height (-0.8∈±∈1.0 SDS, p∈<∈0.001) and not different among the 4 quartiles for birthdate. It was correlated with target height (r2∈=∈0.564, p∈<∈0.001) and height at puberty onset (r2∈=∈0.685, p∈<∈0.001), but not with age at diagnosis or the starting LT4/kg/day dose. The curve fitting analysis showed that the age at diagnosis progressively decreased during the 20-year study period, while the target height and the starting LT4/kg/day increased. Final height was not affected by the birthdate, the age at diagnosis, the starting LT4 dose. Conclusions: The final height is higher than the target height, but despite the improvement in the screening and the treatment, it did not improve over the last 20 years. These findings are in keeping with the described secular trend and suggest that earlier diagnosis and replacement therapy do not significantly modify final height in these patient

Transcriptome based identification of mouse cumulus cell markers that predict the developmental competence of their enclosed antral oocytes

BACKGROUND: The cumulus cells (CCs) enveloping antral and ovulated oocytes have been regarded as putative source of non-invasive markers of the oocyte developmental competence. A number of studies have indeed observed a correlation between CCs gene expression, embryo quality, and final pregnancy outcome. Here, we isolated CCs from antral mouse oocytes of known developmental incompetence (NSN-CCs) or competence (SN-CCs) and compared their transcriptomes with the aim of identifying distinct marker transcripts. RESULTS: Global gene expression analysis highlighted that both types of CCs share similar transcriptomes, with the exception of 422 genes, 97.6% of which were down-regulated in NSN-CCs vs. SN-CCs. This transcriptional down-regulation in NSN-CCs was confirmed by qRT-PCR analysis of CC-related genes (Has2, Ptx3, Tnfaip6 and Ptgs2). Only ten of the 422 genes were up-regulated with Amh being the most up-regulated in NSN-CCs, with an average 4-fold higher expression when analysed by qRT-PCR. CONCLUSIONS: The developmental incompetence (NSN) or competence (SN) of antral oocytes can be predicted using transcript markers expressed by their surrounding CCs (i.e., Has2, Ptx3, Tnfaip6, Ptgs2 and Amh). Overall, the regulated nature of the group of genes brought out by whole transcriptome analysis constitutes the molecular signature of CCs associated either with developmentally incompetent or competent oocytes and may represent a valuable resource for developing new molecular tools for the assessment of oocyte quality and to further investigate the complex bi-directional interaction occurring between CCs and oocyte

Case Report - Multinodular goiter in a patient with Congenital Hypothyroidism and Bannayan-Riley-Ruvalcaba syndrome: the possible synergic role of TPO and PTEN mutation

We report the case of a paediatric female patient affected by Bannayan-Riley-Ruvalcaba syndrome (BRRS) and congenital hypothyroidism (CH) with homozygous mutation of the TPO gene. She underwent total thyroidectomy at the age of seven years because of the development of a multinodular goiter. BRRS patients present an increased risk of benign and malignant thyroid disease since childhood because of inactivating mutation of PTEN, an onco-suppressor gene. Instead, homozygous mutations in the TPO gene can be associated with severe forms of hypothyroidism with goiter; previous studies have described cases of follicular and papillary thyroid cancer in CH patients with TPO mutation despite a perfectly controlled thyroid function with Levothyroxine therapy. To our knowledge, this is the first case that describes the possible synergic role of coexisting mutation of both TPO and PTEN in the development of multinodular goiter underlining the importance of a tailored surveillance program in these patients, especially during childhood

The role of immune suppression in COVID-19 hospitalization: clinical and epidemiological trends over three years of SARS-CoV-2 epidemic

Specific immune suppression types have been associated with a greater risk of severe COVID-19 disease and death. We analyzed data from patients &gt;17 years that were hospitalized for COVID-19 at the “Fondazione IRCCS Ca′ Granda Ospedale Maggiore Policlinico” in Milan (Lombardy, Northern Italy). The study included 1727 SARS-CoV-2-positive patients (1,131 males, median age of 65 years) hospitalized between February 2020 and November 2022. Of these, 321 (18.6%, CI: 16.8–20.4%) had at least one condition defining immune suppression. Immune suppressed subjects were more likely to have other co-morbidities (80.4% vs. 69.8%, p &lt; 0.001) and be vaccinated (37% vs. 12.7%, p &lt; 0.001). We evaluated the contribution of immune suppression to hospitalization during the various stages of the epidemic and investigated whether immune suppression contributed to severe outcomes and death, also considering the vaccination status of the patients. The proportion of immune suppressed patients among all hospitalizations (initially stable at &lt;20%) started to increase around December 2021, and remained high (30–50%). This change coincided with an increase in the proportions of older patients and patients with co-morbidities and with a decrease in the proportion of patients with severe outcomes. Vaccinated patients showed a lower proportion of severe outcomes; among non-vaccinated patients, severe outcomes were more common in immune suppressed individuals. Immune suppression was a significant predictor of severe outcomes, after adjusting for age, sex, co-morbidities, period of hospitalization, and vaccination status (OR: 1.64; 95% CI: 1.23–2.19), while vaccination was a protective factor (OR: 0.31; 95% IC: 0.20–0.47). However, after November 2021, differences in disease outcomes between vaccinated and non-vaccinated groups (for both immune suppressed and immune competent subjects) disappeared. Since December 2021, the spread of the less virulent Omicron variant and an overall higher level of induced and/or natural immunity likely contributed to the observed shift in hospitalized patient characteristics. Nonetheless, vaccination against SARS-CoV-2, likely in combination with naturally acquired immunity, effectively reduced severe outcomes in both immune competent (73.9% vs. 48.2%, p &lt; 0.001) and immune suppressed (66.4% vs. 35.2%, p &lt; 0.001) patients, confirming previous observations about the value of the vaccine in preventing serious disease

Acquisition of the oocyte developmental competence

Oocyte developmental competence: in search of a transcriptional signature Maurizio Zuccotti1, Valeria Merico2, Martina Belli2, Giulia Vigone2, Silvia Garagna2 1. Dipartimento di Scienze Biomediche, Biotecnologiche e Traslazionali (S.Bi.Bi.T.), Sezione di Anatomia, Istologia ed Embriologia, University of Parma, Italy. 2. Dipartimento di Biologia e Biotecnologie ‘Lazzaro Spallanzani’, Laboratorio di Biologia dello Sviluppo, University of Pavia, Italy. In our laboratory we try to unravel the molecular signature that lays behind the developmental competence of a mammalian egg, aiming at finding markers that could help to the evaluation of the quality of the female gamete. Here, we will present a series of studies that led to the identification of transcriptional networks (TN) differentially expressed in developmentally competent or incompetent oocytes, and also we will show that one of these TNs is maintained during preimplantation and in embryonic stem cells (ESCs), representing a fil rouge of developmental continuity that links the egg to the blastocyst. To this end, we made use of a model study in which a type specific of metaphase II (MII) oocyte ceases development at the 2-cell stage. Based on their chromatin organisation – as observed after staining with the Hoechst 33342 supravital fluorochrome - fully-grown mouse antral oocytes are classified into surrounded nucleolus (SN) or not surrounded nucleolus (NSN) oocytes (Debey et al., 1992; Mattson and Albertini, 1990; Zuccotti et al., 1995). Following a different time-course and chromatin rearrangements (Belli et al., 2014) both oocytes mature in vitro to MII (MIISN and MIINSN), but, when fertilised, only MIISN may reach full-term development, whereas MIINSN arrest at the 2-cell stage (Zuccotti et al., 1998, 2005). To understand the developmental incompetence of MIINSN oocytes, we investigated into their transcriptional legacy. Gene expression was compared by whole-transcriptome microarrays analysis, which brought up 380 differentially expressed genes, 77 down-regulated and 303 up-regulated. Further bioinformatics, RT-PCR and immunocytochemistry analyses of these differentially expressed genes and proteins showed an emerging network of 25 genes mostly up-regulated in MIINSN oocytes and assigned to adverse biochemical pathways such as apoptosis and mitochondrial dysfunction (Zuccotti et al., 2008; 2009, 2011a). Importantly, most of these genes are known to be regulated by the transcription factor OCT4, one of a handful of master transcription factors that regulate cell pluripotency. In a next step, we compared the whole-transcriptional profile of developmentally competent vs. incompetent oocytes and that of their derived 2-cell embryos; the OCT4-TN was further expanded to 80 transcripts, mostly expressed in cancer cells and 37 notable companions of the OCT4 transcriptome in ESCs (Zuccotti et al., 2011b; 2012). For the first time, these results indicate that the OCT4-TN may represent a developmental link between eggs, early preimplantation embryos and ESCs, indicating that the molecular signature that characterises the ESCs pluripotency may be rooted in oogenesis. Bibliography Mattson BA, Albertini DF: Oogenesis: chromatin and microtubule dynamics during meiotic prophase. Mol Reprod Dev 25: 374-383, 1990. Debey P, Szöllösi MS, Szöllösi D, Vautier D, Girousse A, Besombes D. Competent mouse oocytes isolated from antral follicles exhibit different chromatin organization and follow different maturation dynamics. Mol Reprod Dev 36: 59-74, 1993. Zuccotti M, Piccinelli A, Giorgi Rossi P, Garagna S, Redi C: Chromatin organization during mouse oocyte growth. Mol Reprod Dev 41: 479-485, 1995. Zuccotti M, Giorgi Rossi P, Martinez A, Garagna S, Forabosco A, Redi CA. Meiotic and developmental competence of mouse antral oocytes. Biol Reprod 58: 700-704, 1998. Zuccotti M, Garagna S, Merico V, Monti M, Alberto Redi C: Chromatin organisation and nuclear architecture in growing mouse oocytes. Mol Cell Endocrinol 234: 11-17, 2005. Zuccotti M, Merico V, Sacchi L, Bellone M, Brink TC, Stefanelli M, Redi CA, Bellazzi R, Adjaye J, Garagna S. Oct-4 regulates the expression of Stella and Foxj2 at the Nanog locus: implications for the developmental competence of mouse oocytes. Hum Reprod. 24: 2225-2237, 2009. Zuccotti M, Merico V, Cecconi S, Redi CA, Garagna S. What does it take to make a developmentally competent mammalian egg? Hum Reprod Update 17: 525-540, 2011a. Zuccotti M, Merico V, Bellone M, Mulas F, Sacchi L, Rebuzzini P, Prigione A, Redi CA, Bellazzi R, Adjaye J, Garagna S. Gatekeeper of pluripotency: a common Oct4 transcriptional network operates in mouse eggs and embryonic stem cells. BMC Genomics 12: 1-13, 2011. Zuccotti M, Merico V, Belli M, Mulas F, Sacchi L, Zupan B, Redi CA, Prigione A, Adjaye J, Bellazzi R, Garagna S. OCT4 and the acquisition of oocyte developmental competence during folliculogenesis. Int J Dev Biol. 56: 853-858, 2012. Belli M, Vigone G, Merico V, Redi CA, Garagna S, Zuccotti M. Time-lapse dynamics of the mouse oocyte chromatin organisation during meiotic resumption. Biomed Res Int. 2014:207357, 2014

Time-Lapse Dynamics of the Mouse Oocyte Chromatin Organisation during Meiotic Resumption

In the mammalian oocyte, distinct patterns of centromeres and pericentromeric heterochromatin localisation correlate with the gamete’s developmental competence. Mouse antral oocytes display two main types of chromatin organisation: SN oocytes, with a ring of Hoechst-positive chromatin surrounding the nucleolus, and NSN oocytes lacking this ring. When matured to MII and fertilised, only SNoocytes develop beyond the 2-cell, and reach full term. To give detailed information on the dynamics of the SN or NSN chromatin during meiosis resumption, we performed a 9 hr time-lapse observation.Themain significant differences recorded are: (1) reduction of the nuclear area only in SN oocytes; (2) ∼17 min delay of GVBD in NSN oocytes; (3) chromatin condensation, after GVBD, in SN oocytes; (4) formation of 4-5 CHCs in SN oocytes; (5) increase of the perivitelline space, ∼57 min later in NSN oocytes; (6) formation of a rosette-like disposition of CHCs, ∼84 min later in SN oocytes; (7) appearance of the MI plate ∼40 min later in NSN oocytes. Overall, we described a pathway of transition from the GV to the MII stage that is punctuated of discrete recordable events showing their specificity and occurring with different time kinetics in the two types of oocytes

Towards a 3D culture of mouse ovarian follicles

The ovarian follicle has a three-dimensional (3D) structure in which the oocyte is surrounded by tightly connected follicle cells that mediate the action of external signals and nourish the gamete during its maturation. Thus, the maintenance of the follicle organization during the whole growth process is crucial for the correct acquisition of the developmental competence. In recent years, much attention has been given to in vitro culture systems capable of maintaining the follicle architecture. With the aim of providing a quick reference guide, in this review we will summarize the techniques developed for the 3D culture of mouse follicles