Follicular Tissue Research Articles

8049 Beyond Metastasis: Parasitic Thyroid Tissue Mimicking Thyroid Cancer

Abstract Disclosure: M.S. Shah: None. S. Humayon: None. N.A. Mungo: None. This case study details a complex clinical scenario involving a patient with diaphoresis and a familial history of thyroid cancer. Initial suspicions of functionally active metastatic thyroid cancer led to a thyroidectomy and cervical lymph node dissection, only to reveal unexpected parasitic thyroid tissue. This case highlights the importance of meticulous evaluation and follow up similar presentations, shedding light on the rarity of this phenomenon.The patient, experiencing a year of diaphoresis unresponsive to estradiol therapy, underwent imaging revealing multiple thyroid nodules. A left superior nodule (TI-RADS 4) and a more concerning right mid-posterior nodule (TI-RADS 5) prompted fine needle aspiration (FNA). FNA of the right thyroid nodule exhibited Bethesda category III atypia, no molecular samples were taken at that time, while cervical lymph node FNA yielded nondiagnostic results.Subsequent ultrasound showed no interval change in the right thyroid nodule but raised concerns about increased solid components and decreased echogenicity in other nodules. Due to patient concerns of positive family history of thyroid cancer, the lymph node was biopsied. The result yielded a low cellular sample, but follicular thyroid tissue was present. The patient elected to undergo thyroidectomy as she did not want to continue monitoring. Pathology revealed parasitic thyroid tissue in the lymph node and nodular follicular thyroid disease, with no evidence of papillary thyroid carcinoma in the gland. Notably, the patient had no history of thyroid surgeries, ruling out traumatic rupture or implantation from a follicular adenoma. However, the possibility of follow-up metastatic follicular carcinoma cannot be entirely excluded on histologic grounds.Follow-up tests were recommended, encompassing mutational analysis, whole-body scans, and thyroglobulin (Tg) monitoring. Molecular analysis did not favor cancer. Tg levels remained at (18 ng/ml, n< 35ng/ml) of initial evaluation six weeks post surgery and repeat testing is pending. Ablative radioactive iodine therapy may be appropriate as resection of all parasitic thyroid tissue can prove difficult.This case emphasizes the diagnostic challenges posed by the rare occurrence of parasitic thyroid tissue, often mistaken for metastatic thyroid cancer. The findings contribute to our understanding of uncommon thyroid pathologies. The clinical implications of this case extend to the importance of heightened clinical suspicion in intricate endocrine cases. Presentation: 6/3/2024

New insights into the ovulatory process in the human ovary.

Successful ovulation is essential for natural conception and fertility. Defects in the ovulatory process are associated with various conditions of infertility or subfertility in women. However, our understanding of the intra-ovarian biochemical mechanisms underlying this process in women has lagged compared to our understanding of animal models. This has been largely due to the limited availability of human ovarian samples that can be used to examine changes across the ovulatory period and delineate the underlying cellular/molecular mechanisms in women. Despite this challenge, steady progress has been made to improve our knowledge of the ovulatory process in women by: (i) collecting granulosa cells across the IVF interval, (ii) creating a novel approach to collecting follicular cells and tissues across the periovulatory period from normally cycling women, and (iii) developing unique in vitro models to examine the LH surge or hCG administration-induced ovulatory changes in gene expression, the regulatory mechanisms underlying the ovulatory changes, and the specific functions of the ovulatory factors. The objective of this review is to summarize findings generated using in vivo and in vitro models of human ovulation, with the goal of providing new insights into the mechanisms underlying the ovulatory process in women. This review is based on the authors' own studies and a search of the relevant literature on human ovulation to date using PubMed search terms such as 'human ovulation EGF-signaling', 'human ovulation steroidogenesis', 'human ovulation transcription factor', 'human ovulation prostaglandin', 'human ovulation proteinase', 'human ovulation angiogenesis' 'human ovulation chemokine', 'human ovulatory disorder', 'human granulosa cell culture'. Our approach includes comparing the data from the authors' studies with the existing microarray or RNA-seq datasets generated using ovarian cells obtained throughout the ovulatory period from humans, monkeys, and mice. Current findings from studies using in vivo and in vitro models demonstrate that the LH surge or hCG administration increases the expression of ovulatory mediators, including EGF-like factors, steroids, transcription factors, prostaglandins, proteolytic systems, and other autocrine and paracrine factors, similar to those observed in other animal models such as rodents, ruminants, and monkeys. However, the specific ovulatory factors induced, their expression pattern, and their regulatory mechanisms vary among different species. These species-specific differences stress the necessity of utilizing human samples to delineate the mechanisms underlying the ovulatory process in women. The data from human ovulation in vivo and in vitro models have begun to fill the gaps in our understanding of the ovulatory process in women. Further efforts are needed to discover novel ovulatory factors. One approach to address these gaps is to improve existing in vitro models to more closely mimic in vivo ovulatory conditions in humans. This is critically important as the knowledge obtained from these human studies can be translated directly to aid in the diagnosis of ovulation-associated pathological conditions, for the development of more effective treatment to help women with anovulatory infertility or, conversely, to better manage ovulation for contraceptive purposes. N/A.

Local Regulatory Changes of HSD11B1 and NR3C1 in the Follicular and Luteal Tissue During Experimentally Induced Ovulation in the Cow.

The objective of the study was to characterise the expression patterns of the two key components of cortisol action namely HSD11B1 (11-beta-hydroxysteroid dehydrogenase type 1) and NR3C1 (nuclear receptor subfamily 3, group C, member 1, also known as the glucocorticoid receptor) in superovulation induced bovine follicles during the periovulation and subsequent corpus luteum (CL) formation. Bovine ovaries containing preovulatory follicles or CL were timely defined during induced ovulation as follows: 0 h before GnRH (Gonadotropin-releasing hormone) application, and 4, 10, 20, 25 (follicles) and 60 h (early CL) after GnRH. The low mRNA expression of HSD11B1 and NR3C1 in the follicle group before the GnRH application increased significantly in the follicle group 20 h after GnRH and remained high afterward also in the early CL group. In contrast, the high NR3C1 mRNA decreased in follicles 25 h after GnRH (close to ovulation) and significantly increased again after ovulation (early CL). Our results indicated the involvement of HSD11B1 and NR3C1 as the two key components of cortisol action in the local mechanisms coordinating final follicle maturation, ovulation, follicular-luteal transition and CL development in the cow.

A Novel Treatment of Acne Vulgaris Using a 1927 nm Fractional Thulium Laser: A Case Series.

Acne vulgaris remains one of the most common and problematic dermatological conditions. Recently, a fractional 1927 nm thulium laser has been developed with specific water absorption characteristics which may be of interest in the treatment of acne. Nine consecutive Korean subjects, 6 females and 3 males, ages ranging from 13 to 33 yr, presented with a mixture of inflammatory and noninflammatory acne. Baseline clinical photography, image analysis and lesion counts were performed. A fractional 1927 nm thulium laser (FTL) delivered 6 treatment sessions in 5 subjects and 5 sessions in 4 subjects, 4 weeks between sessions. Pain during treatment was assessed. At 32 weeks after the last treatment session, an independent Investigator Global Assessment (IGA) performed lesion counts and graded the severity of the acne at baseline and the final assessment on a quintile scale. Data were analyzed statistically. All 9 subjects completed the study with significant reductions in the inflammatory and noninflammatory lesions (P values 0.0012 and 0.0081, respectively) with overall lesion counts at the final assessment ranging from 60% to 97.1%, and acne grades in the IGA dropping by an average of 1.67 (range 1 to 3 grades). There was no significant difference in lesion counts or acne grades between the subjects who had 6 treatments and those who had 5 (P = 0.7695). Mild pain was reported during treatment, and no adverse events were reported by either the subjects or investigator. The FTL at the parameters used in the present study caused disruption to the upper portion of the affected follicles and sebaceous glands under an intact stratum corneum, thereby destroying or damaging the causative Cutibacterium acnes. The superficial controlled coagulation additionally induced follicular remodeling and tissue regeneration, potentially contributing to the noticeable results in inflammatory and noninflammatory acne lesions.

Age-related changes in the mitochondrial, synthesis of steroids, and cellular homeostasis of the chicken ovary

In poultry reproduction, the decline of ovarian function due to aging is related to dysfunction of mitochondria exacerbated by a reduction in antioxidant capacity, ultimately leading to follicle atresia and decreased egg production. However, the mechanisms of mitochondrial dysfunction in the chicken ovary in aging have remained to be understood. Hence, this study aims to investigate the effects of aging on mitochondrial function and cellular homeostasis. We collect ovarian tissue, small white follicles (SWF), large white follicles (LWF), and small yellow follicles (SYF) from three different laying periods of hens. The transmission electron microscopy (TEM) results showed that mitochondrial damage occurred in ovarian tissue during the late laying period (LP), characterized by structural swelling, scattered mitochondrial cristae, and an increase in the vacuoles. At the same time, with age, the synthesis of steroid hormones in the ovaries and follicular tissues is reduced. The levels of autophagy and cell apoptosis in ovarian tissues were both increased in the LP. In addition, aging adversely impacts mitochondrial function, leading to a decrease in mitochondrial unfolded protein response (UPRmt) functions. This study will expand the knowledge about regressing ovarian aging in hens and increasing egg production in older layers for poultry production.

Optimizing decellularization protocols for human thyroid tissues: a step towards tissue engineering and transplantation

Hypothyroidism is caused by insufficient stimulation or disruption of the thyroid. However, the drawbacks of thyroid transplantation have led to the search for new treatments. Decellularization allows tissue transplants to maintain their biomimetic structures while preserving cell adhesion, proliferation, and differentiation. This study aimed to decellularize human thyroid tissues using a structure-preserving optimization strategy and present preliminary data on recellularization. Nine methods were used for physical and chemical decellularization. Quantitative and immunohistochemical analyses were performed to investigate the DNA and extracellular matrix components of the tissues. Biomechanical properties were determined by compression test, and cell viability was examined after seeding MDA-T32 papillary thyroid cancer (PTC) cells onto the decellularized tissues. Decellularized tissues exhibited a notable decrease (<50 ng mg−1 DNA, except for Groups 2 and 7) compared to the native thyroid tissue. Nonetheless, collagen and glycosaminoglycans were shown to be conserved in all decellularized tissues. Laminin and fibronectin were preserved at comparatively higher levels, and Young’s modulus was elevated when decellularization included SDS. It was observed that the strain value in Group 1 (1.63 ± 0.14 MPa) was significantly greater than that in the decellularized tissues between Groups 2–9, ranging from 0.13 ± 0.03–0.72 ± 0.29 MPa. Finally, viability assessment demonstrated that PTC cells within the recellularized tissue groups successfully attached to the 3D scaffolds and sustained metabolic activity throughout the incubation period. We successfully established a decellularization optimization for human thyroid tissues, which has potential applications in tissue engineering and transplantation research. Our next goal is to conduct recellularization using the methods utilized in Group 1 and transplant the primary thyroid follicular cell-seeded tissues into an in vivo animal model, particularly due to their remarkable 3D structural preservation and cell adhesion-promoting properties.

Investigation into the association of FNDC1 and ADAMTS12 gene expression with plumage coloration in Muscovy ducks.

To elucidate the molecular genetic mechanisms underpinning feather color in Muscovy ducks. A cohort of 100 Muscovy ducks was meticulously selected for this research. Follicular tissues from ducks exhibiting black and white plumage served as the experimental samples. From these tissues, RNA and proteins were extracted for further analysis. The RNA underwent reverse transcription polymerase chain reaction amplification, followed by validation through western blot assays. The data revealed a significant upregulation in the expression of FN domain-containing protein 1 (FNDC1) and ADAMTS12 genes in Muscovy ducks with white plumage traits as opposed to those with black plumage traits. Specifically, individuals with pure white plumage demonstrated a markedly elevated expression of the FNDC1 gene in comparison to their pure black counterparts. Conversely, expression levels of the ADAMTS12 gene were found to be reduced in ducks with pure black plumage relative to those with pure white plumage. Notably, the expression patterns of FNDC1 and ADAMTS12 genes exhibited inconsistencies between mRNA and protein levels. This study offers significant insights into the molecular genetic mechanisms underlying feather color variation in Muscovy ducks. FNDC1 and ADAMTS12 could be considered potential targets for genetic manipulation or selective breeding strategies aimed at achieving specific feather color phenotypes in Muscovy ducks.

Neuroectodermal influence in odontogenic cyst and tumor: evidence through CD56 immunoexpression.

CD56, associated with neuroectodermal differentiation of the embryonal cells, is often considered a marker of neural lineage. Odontogenic keratocysts (OKCs) are of particular interest because of their characteristic histopathologic features, high recurrence rate, and aggressive behavior. CD56 immunoreactivity in these lesions has been reported with very high frequency. The present study analyzes the immunoexpression of CD56 in ameloblastoma (AM) and OKC to infer neuroectodermal influence in the pathogenesis of odontogenic lesions and its correlation with clinicopathologic parameters. Fifty histopathologically confirmed cases of OKC and AM, 25 from tooth-bearing (TB) and molar-ramus (MR) regions each, and 5 dental follicular tissues as control were collected from the department archives and immunohistochemical analysis with CD56 was carried out. CD56 immunopositivity was seen in 64% AM and 36% OKC cases. The majority of AM cases showed cytoplasmic expression in the peripheral cells of odontogenic islands; similarly, OKC cases showed continuous and uniform cytoplasmic expression in the basal and parabasal cells of the cystic lining. CD56 immunopositivity was found in more AM cases as compared to OKC cases in both the TB and MR regions. The assessment of CD56 immunoexpression in odontogenic cyst and tumor (AM) may aid in understanding the role of neuroectodermal influence in the etiopathogenetic pathways and a possible influence of CD56 on the clinical behavior and aggressiveness of the odontogenic lesions. A correlation of CD56 expression with the clinical outcome of the disease (site, perforation, root resorption, and tooth displacement) can help envisage possible prognostic assessment for these lesions.

ADPN Regulates Oxidative Stress-Induced Follicular Atresia in Geese by Modulating Granulosa Cell Apoptosis and Autophagy.

Geese are susceptible to oxidative stress during reproduction, which can lead to follicular atresia and impact egg production. Follicular atresia is directly triggered by the apoptosis and autophagy of granulosa cells (GCs). Adiponectin (ADPN), which is secreted by adipose tissue, has good antioxidant and anti-apoptotic capacity, but its role in regulating the apoptosis of GCs in geese is unclear. To investigate this, this study examined the levels of oxidative stress, apoptosis, and autophagy in follicular tissues and GCs using RT-qPCR, Western blotting, immunofluorescence, flow cytometry, transcriptomics and other methods. Atretic follicles exhibited high levels of oxidative stress and apoptosis, and autophagic flux was obstructed. Stimulating GCs with H2O2 produced results similar to those of atretic follicles. The effects of ADPN overexpression and knockdown on oxidative stress, apoptosis and autophagy in GCs were investigated. ADPN was found to modulate autophagy and reduced oxidative stress and apoptosis in GCs, in addition to protecting them from H2O2-induced damage. These results may provide a reasonable reference for improving egg-laying performance of geese.

The Role of Mesenchymal Stem Cells Secretome in Regeneration of Thyroid Tissue Damage after Thyroid Lobectomy: A Literature Review

Mesenchymal Stem Cells (MSC) are a type of stem cell that is multipotent and originates from the mesoderm and ectoderm at an early stage of development and can differentiate into: 1. Ectodermal derivatives such as: neurons, keratocytes and keratinocytes; 2. Endodermal derivatives such as: hepatocytes and pancreatic β cells. MSCs were first discovered in the bone marrow, but in recent research they have been found in many other body tissues, such as the lungs, pancreas, liver, adipose tissue, synovial tissue, muscles, bones, amniotic fluid and umbilical cord blood. MSCs vary in content, accessibility, proliferation ability, immunomodulatory capacity, and the cytokines they secrete, and have different therapeutic potential based on different sources. Recent evidences showed that even the secreted factor including secretome, microvesicles, or exosomes alone without the cell may repair damaged tissue in various conditions. In this review, we summarize the latest research which aims to provide an overview of the influence of the mesenchymal stem cells secretome in regenerating damaged follicular thyroid tissue after thyroid lobectomy, so that in the future research can be carried out on this matter to minimize complications after thyroid lobectomy, can even regenerate follicular thyroid tissue, and can be an option treatment to help overcome the current challenges of hormone replacement therapy.

Does a Single Episode of Pain from Impacted Third Molars Necessitate Their Removal?

Pericoronitis induces oxidative stress in the dental follicular (DF) tissue, which can lead to histopathological alterations. It is a fact that an impacted tooth with a history of recurrent pericoronitis should be extracted. But there is a limited research on the levels of nitric oxide (NO) in DF tissues of impacted mandibular third molars with even a single episode of pain, necessitating their removal biochemically at a younger age. A cross-sectional study was designed to assess and compare the levels of nitric oxide in DF tissues of impacted mandibular third molars without a history of pain to those with a history of a single episode of pain in the past six months. Sixty individuals with impacted third molars were enrolled in a single-blind cross-sectional study. Group A (n = 30) had patients with a history of single episode of pain in the last six months, and Group B (n = 30) had patients with no history of pain. Impacted third molar extractions were carried out for consenting individuals in the concerned groups. Dental follicle tissue was collected to assess their nitric oxide levels through a spectrophotometric assay biochemically. Nitric oxide levels were significantly higher for patients with a history of a single episode of pain in the past six months than those without any history of pain (p < 0.030). The findings of this study suggest that NO could be used as an indicator of oxidative stress and therefore emphasizes the importance of removing impacted mandibular third molars in patients who have had even a single episode of pain.

NR5A1 and NR5A2 regulate follicle development in chicken (Gallus gallus) by altering proliferation, apoptosis, and steroid hormone synthesis of granulosa cells

Chicken ovarian follicle development is regulated by complex and dynamic gene expression. Nuclear receptor 5A1 and 5A2 (NR5A1 and NR5A2, respectively) are key genes that regulate steroid hormone production and gonadal development in mammals; however, studies on follicular development in the chicken ovary are scarce. In this study, we investigated the functions of NR5A1 and NR5A2 on follicle development in chickens. The results showed that the expression of NR5A1 and NR5A2 was significantly higher in small yellow follicles and F5. Furthermore, the expression of NR5A1 and NR5A2 was significantly higher in follicular tissues of peak-laying hens (30 wk) than in follicular tissues of late-laying hens (60 wk), with high expression abundance in granulosa cells (GC). The overexpression of NR5A1 and NR5A2 significantly promoted proliferation and inhibited apoptosis of cultured GC; upregulated STAR, CYP11A1, and CYP19A1 expression and estradiol (E2) and progesterone (P4) synthesis in GC from preovulatory follicles (po-GC); and increased STAR, CYP11A1, and CYP19A1 promoter activities. In addition, follicle-stimulating hormone treatment significantly upregulated NR5A1 and NR5A2 expression in po-GC and significantly promoted FSHR, CYP11A1, and HSD3B1 expression in GC from pre-hierarchical follicles and po-GC. The core promoter region of NR5A1 was identified at the -1,095- to -483-bp and -2,054- to -1,536-bp regions from the translation start site (+1), and the core promoter region of NR5A2 was at -998 to -489 bp. Two single nucleotide polymorphisms (SNP) were identified in the core promoter region of the NR5A1 gene, which differed between high- and low-yielding chicken groups. Our study suggested that NR5A1 and NR5A2 promoted chicken follicle development by promoting GC proliferation and E2 and P4 hormone synthesis and inhibiting apoptosis. Moreover, we identified the promoter core region or functional site that regulates NR5A1 and NR5A2 expression.

EphB3 protein is a potential ancillary diagnostic biomarker for thyroid cancers

ObjectiveTo investigate the expression of ephrin type B receptor 3 (EphB3) in thyroid tumors and its usage as an ancillary diagnostic biomarker for thyroid tumors. MethodsFormalin-fixed and paraffin-embedded (FFPE) tissue samples (78 cases) and FNAC samples (57 cases) were assessed with the EphB3 antibody using immunohistochemistry. PTC and other thyroid follicular tumors were compared regarding their EphB3 expression. Sanger sequencing was used to assess for the presence of a BRAF V600E mutation. ResultsEphB3 was positive in 81.8 % (27/33) of papillary thyroid carcinoma (PTC), 83.3 % (5/6) of medullary thyroid carcinoma (MTC), 25 % (1/4) of hyperplastic/adenomatoid nodule (HN), 14.3 % (1/7) of follicular adenoma (FA), and negative in follicular tumors of uncertain malignant potential (FT-UMP) (0/13), noninvasive follicular neoplasm with papillary-like nuclear features (NIFTP) (0/7), thyroid follicular carcinoma (TFC) (0/4), Hashimoto's thyroiditis (0/4), and normal thyroid follicular tissues (0/33). In cellular blocks, EphB3 was positive in 87.1 % (20/23) of PTC, 75 % (3/4) of MTC, 20 % (2/10) of HN, and negative in atypia of undetermined significance/follicular lesion of undetermined significance (AUS/FLUS) (0/20) and normal thyroid follicular cells (0/10). ConclusionEphB3 is expressed in the majority of PTC, but less so in benign follicular nodules. EphB3 expression in fine needle aspiration cytology (FNAC) specimens can be used as a diagnostic tool to differentiate thyroid cancer from other follicular lesions in its differential diagnosis, especially AUS/FLUS and PTC.

PTEN inhibitor and kit ligand increase in vitro activation and survival of primordial follicles in alpaca.

In mammals, activation of primordial follicles to primary follicle is a progressive and highly regulated process. There is evidence in mice that phosphatase and tensin homologue deleted on Chromosome 10 (PTEN) silencing is an important negative regulator of phosphatidylinositol 3-kinase (PI3K), which initiates activation of dormant follicles. The objective of the study was to evaluate the effect of the addition of PTEN inhibitor (bpV(HOpic)) (10 μM) and/or Kit Ligand (KL) (100 ng/mL) on the in vitro activation and survival of alpaca primordial follicles. Ovarian cortical fragments from 11 adult alpacas were cultured for 24 h in tissue culture medium (α-MEM+ ) supplemented with KL and bpV or the association of both. Subsequently, each sample was processed by classical histology and follicular counting and classification were performed. The results obtained show a reduction (p < 0.05) of primordial follicles in more than 50% in follicular tissue cultured in vitro in α-MEM+ or supplemented with bpV and/or KL versus the control (not cultured). Further, >25% increase in primary follicles in follicular tissue cultured in vitro in α-MEM+ or supplemented with KL and/or bpV versus control. However, the follicular survival rate showed a decrease of 20% in the cultured tissues, except for the α-MEM+ supplemented with KL and bpV. In conclusion, supplementation of bpV (HOpic) (10 μM) and KL (100 ng/mL) increased the activation in vitro of primordial follicles and survival after in vitro culture of alpaca ovarian tissue.

Thyroiditis – A Clinical Update and Review

Purpose: Thyroiditis refers to a set of inflammatory disorders involving the destruction of normal thyroid follicular tissue. Each disorder has distinctive histology and pathology. Understanding each condition in the thyroiditis frame is crucial for physicians. Main: This review categorizes thyroiditis into two groups based on the presence or absence of tenderness: painful and painless. This paper reviews the primary etiologies, diagnostic modalities, and treatment options for each condition. The painful thyroiditis comprises subacute, infectious, radiation, trauma-induced thyroiditis, and rarely Hashimoto’s thyroiditis. The painless group consists of subacute lymphocytic thyroiditis, postpartum thyroiditis, drug-induced thyroiditis, and fibrous thyroiditis. Findings: In the painful group, the primary etiology of subacute thyroiditis is viral, including SARS-CoV- 2, which has been reported recently, and the main etiology of the infectious subgroup is bacterial infections. Symptom management and pain relief are the mainstays of treatment for painful conditions. The painless group typically progresses from transient hyperthyroidism to euthyroidism to hypothyroidism before resolving. Autoimmune and genetics, HLA-DR3, likely contribute to subacute lymphocytic thyroiditis. Certain medications, including interferon-alpha (HVC management), IL2 Tyrosine-kinase inhibitors (cancer management), amiodarone, lithium, and check-point inhibitor immunotherapy (CTLA-4 and PCD-1) are found to be related to thyroiditis. Fibrous thyroiditis is usually associated with systemic fibrous disease. Conclusion: A comprehensive understanding of each thyroiditis condition’s etiology and clinical presentation is important to accurately diagnose, appropriately manage, and counsel patients on the risk for permanent hypothyroidism that may require long-term thyroid replacement therapy

Detection of La Crosse Virus In Situ and in Individual Progeny to Assess the Vertical Transmission Potential in Aedes albopictus and Aedes aegypti.

La Crosse virus (LACV) is circulating in the midwestern and southeastern states of the United States and can cause human encephalitis. The main vector of the virus is the eastern tree-hole mosquito, Aedes triseriatus. Ae. albopictus has been also described as a natural LACV vector, while Ae. aegypti has been infected with the virus under laboratory conditions. Here, we compare the vertical transmission potential of LACV in Ae. albopictus and Ae. aegypti, with emphasis given to the ovarian infection patterns that the virus generates in both species. Both mosquito species received artificial bloodmeals containing LACV. At defined time points post-infection/bloodmeal, midguts, head tissue, and ovaries were analyzed for the presence of virus. Viral infection patterns in the ovaries were visualized via immunofluorescence confocal microscopy and immunohistopathology assays using an LACV-specific monoclonal antibody. In Ae. aegypti, LACV was confronted with midgut infection and escape barriers, which were much less pronounced in Ae. albopictus, resulting in a significantly higher prevalence of infection in the latter. Following the ingestion of a single virus-containing bloodmeal, no progeny larvae were found to be virus-infected. Regardless, females of both species showed the presence of LACV antigen in their ovariole sheaths. Furthermore, in a single Ae. albopictus female, viral antigen was associated with the nurse cells inside the primary follicles. Following the ingestion of a second non-infectious bloodmeal at 7- or 10-days post-ingestion of an LACV-containing bloodmeal, more progeny larvae of Ae. albopictus than of Ae. aegypti were virus-infected. LACV antigen was detected in the egg chambers and ovariole sheaths of both mosquito species. Traces of viral antigen were also detected in a few oocytes from Ae. albopictus. The low level of vertical transmission and the majority of the ovarian infection patterns suggested the transovum rather than transovarial transmission (TOT) of the virus in both vector species. However, based on the detection of LACV antigen in follicular tissue and oocytes, there was the potential for TOT among several Ae. albopictus females. Thus, TOT is not a general feature of LACV infection in mosquitoes. Instead, the TOT of LACV seems to be dependent on its particular interaction with the reproductive tissues of a female.

Differentiating Thyroid Follicular Adenoma from Follicular Carcinoma via G-Protein Coupled Receptor-Associated Sorting Protein 1 (GASP-1).

Follicular neoplasms are classified as benign or malignant depending on the presence or absence of capsular and/or vascular invasion. Due to incomplete capsular penetration or equivocal vascular invasion, the evaluation of these features can be challenging using histologic examination. In the current study, we analyzed the involvement of G-protein coupled receptor-associated sorting protein 1 (GASP-1) in the development and progression of thyroid neoplasms. Affinity-purified anti-GASP-1 polyclonal antibodies were used for routine immunohistochemistry (IHC) analysis. Thyroid tissue microarrays containing normal thyroid tissue, follicular adenoma, follicular carcinoma, papillary thyroid carcinoma, and anaplastic carcinoma were analyzed. We found that the level of GASP-1 expression can differentiate follicular adenoma from follicular carcinoma. When numerous cases were scored for GASP-1 expression by a board-certified pathologist, we found that GASP-1 expression is 7-fold higher in thyroid malignant neoplasms compared to normal thyroid tissue, and about 4-fold higher in follicular carcinoma compared to follicular adenoma. In follicular adenoma tissues, we observed the presence of many mini-glands that are enriched in GASP-1 and some mini-glands contain as few as three cells. GASP-1 IHC also possesses several advantages over the conventional H&E and can be used to identify early thyroid cancer and monitor cancer progression.

P-219 Single-follicle spatial proteomics of the whole ovary by MALDI mass spectrometry imaging: towards the identification of protein-markers of the follicle's quality

Abstract Study question Can we improve our knowledge of the single-follicle proteome signature by MALDI mass spectrometry imaging? Summary answer We identified follicle-type proteins that function during follicle growth (NUMA1, TPM2), GV-to-MII transition (SFPQ, ACTBL, MARCS, NUCL), ovulation (GELS, CO1A2) and preimplantation development (TIF1B, KHDC3). What is known already The acquisition of oocytes developmental competence occurs through a bidirectional exchange of molecular information between the oocyte, the companion follicle cells, the stroma, and the vascular network. This reciprocal relationship is still poorly understood and particularly scarce is our knowledge of the proteins that are playing a regulative role during folliculogenesis. Matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) is a powerful tool that allows the study of the proteomic landscape of small morphological structures inside a tissue, but in the field of reproduction has been only used to analyse rodent spermatogenesis. Study design, size, duration Nano-scale liquid chromatography-electrospray ionisation-tandem mass spectrometry (nLC-ESI-MS/MS) was combined with MALDI-MSI to identify the proteomic landscape and to map the changes occurring throughout folliculogenesis. We performed single-follicle spatial proteomics on all the follicles, from the secondary to the fully-grown preovulatory, present in histological serial sections of an entire prepubertal 25-day-old mouse ovary. Cross-referencing our results with those obtained by previous proteomics and transcriptomics analyses allowed to strengthen their significance. Participants/materials, setting, methods A single 25-day-old mouse ovary was fixed in 10% formalin, dehydrated, embedded in paraffin, and sectioned to obtain 133 6-μm serial sections. After paraffin removal, trypsin digestion and matrix deposition, mass spectra were acquired using a rapifleX MALDI Tissuetyper™ (Bruker) with a raster sampling of 20 μm in both x and y axes. Then, tryptic peptides were extracted from sections and analysed with nLC-ESI-MS/MS using a Dionex-UltiMate-3000 LC-nano-system coupled with an Impact HD™ UHR-QqToF (Bruker). Main results and the role of chance This study proposes a spatial proteomics workflow to investigate the proteome of a whole prepubertal 25-day-old mouse ovary, preserving the spatial relationship of the peptides in this complex histological context. A total of 401 proteins were identified by nLC-ESI-MS/MS, 69 with a known function in ovary biology. Enrichment analysis highlighted significant KEGG and Reactome pathways, with apoptosis, developmental biology, PI3K-Akt, epigenetic regulation of gene expression, and extracellular matrix organisation being well represented. Then, correlating these data with the spatial information provided by MALDI-MSI on 276 follicles highlighted 94 proteins that were detected throughout the secondary to the pre-ovulatory transition. Of these, 37 proteins showed a gradual quantitative change during follicle differentiation, comprising 10 with a known role in follicle growth (NUMA1, TPM2), oocyte GV-to-MII transition (SFPQ, ACTBL, MARCS, NUCL), ovulation (GELS, CO1A2) and preimplantation development (TIF1B, KHDC3). The proteome landscape identified includes molecules of known function in the ovary, but also those whose specific role is emerging. Altogether, this work demonstrates the utility of performing spatial proteomics in the context of the ovary and offers sound bases for more in-depth investigations that aim to further unravel its spatial proteome. Limitations, reasons for caution A limitation of MALDI-MSI is its 20 µm/pixel resolution which 1) leads to the acquisition of spectra comprising follicular and extra-follicular tissue, particularly with small follicles (15-30µm in diameter); and 2) does not distinguish among the different follicular cell types, thus precluding its application for single-cell in situ proteomics. Wider implications of the findings MALDI-MSI is a potent technology to study the proteome of ovary specimens, but it could also be used for the analysis of their lipidic or metabolic profiles. Our pipelline aim to identify protein-markers of the follicle's quality and, in turn, of the oocyte's developmental competence. Trial registration number not appplicable

Accurate demarcation of a biased nucleus from H&E-stained follicular lymphoma tissues samples

ABSTRACT In recent days, automated demarcation of biased nuclei of follicular lymphoma has become a standard framework in the pipeline of quantitative histopathology. It has received substantial consideration due to subjective variability between different oncologists. This difference can occasionally result in erroneous conclusions, distinct prognostic reports, and inconsistent treatment. This paper provides additional input to an oncologist to ease the prognosis process. This approach defines a local criterion fitting function in neighbourhood of each point based on image intensity. An assumption is pixel intensity will remain constant next to the point. Integration of these local neighbourhood centres leads us to define the global criterion of image segmentation. Segmentation accuracy is evaluated using region-based measures and the optimal segmentation accuracy of 98.6% is achieved using the DICE coefficient. By using locally formulated energy, execution time is also reduced in comparison to the contour algorithm. This technique is independent of the initialization.

Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model.

In recent years, Hashimoto's thyroiditis (HT) has become the most common autoimmune thyroid disease. It is characterized by lymphocyte infiltration and the detection of specific serum autoantibodies. Although the potential mechanism is still not clear, the risk of Hashimoto's thyroiditis is related to genetic and environmental factors. At present, there are several types of models of autoimmune thyroiditis, including experimental autoimmune thyroiditis (EAT) and spontaneous autoimmune thyroiditis (SAT). EAT in mice is a common model for HT, which is immunized with lipopolysaccharide (LPS) combined with thyroglobulin (Tg) or supplemented with complete Freund's adjuvant (CFA). The EAT mouse model is widely established in many types of mice. However, the disease progression is more likely associated with the Tg antibody response, which may vary in different experiments. SAT is also widely used in the study of HT in the NOD.H-2h4 mouse. The NOD.H2h4 mouse is a new strain obtained from the cross of the nonobese diabetic (NOD) mouse with the B10.A(4R), which is significantly induced for HT with or without feeding iodine. During the induction, the NOD.H-2h4 mouse has a high level of TgAb accompanied by lymphocyte infiltration in the thyroid follicular tissue. However, for this type of mouse model, there are few studies to comprehensively evaluate the pathological process during the induction of iodine. A SAT mouse model for HT research is established in this study, and the pathologic changing process is evaluated after a long period of iodine induction. Through this model, researchers can better understand the pathological development of HT and screen new treatment methods for HT.