Clinical heterogeneity and reduced penetrance in DICER1 syndrome: a report of three families

Introduction: DICER1 syndrome is characterized by increased susceptibility to malignancies, mostly occurring in childhood. The range of phenotypic effects of DICER1 variants is under investigation, and the syndrome’s phenotypic spectrum is steadily widening. We report on three Italian families showing heterogeneous clinical presentation and reduced penetrance in family members. Case descriptions: Patient 1 is a 10-year-old girl with a Sertoli-Leydig cell tumor. Although family history was unremarkable, genetic testing identified a DICER1 germline variant, inherited from her healthy father. Benign thyroid nodules were subsequently diagnosed in both the proband and her father. Patient 2 is an 8-month-old boy with type 1 pleuropulmonary blastoma. His sister developed a nephroblastoma at age 2 years. A DICER1 novel variant was identified in both siblings and their healthy father. Patient 3 is a 22-year-old man who developed a spinal extramedullary intradural mass diagnosed as rhabdomyosarcoma with a peculiar tubular, gland-like component. Tumor testing revealed two pathogenic DICER1 variants, one of which was confirmed to be germline and identified in his 17-year-old healthy brother and in his father, who showed multiple thyroid nodules. Conclusions: Among our patients, three developed tumors most frequently associated with DICER1 syndrome (i.e. pleuropulmonary blastoma, nephroblastoma, and Sertoli-Leydig cell tumor). One developed a peculiar sarcoma of the spinal cord not previously described in DICER1 syndrome. Genetic testing in relatives highlighted the paternal origin and reduced penetrance in all families, with thyroid benign lesions as the most common features in otherwise unaffected individuals.

Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens

Background miRNAs regulate circadian patterns by modulating the biological clocks of animals. In our previous study, we found that the clock gene exhibited a cosine expression pattern in the fallopian tube of chicken uterus. Clock-controlled miRNAs are present in mammals and Drosophila; however, whether there are clock-controlled miRNAs in the chicken uterus and, if so, how they regulate egg-laying rhythms is unclear. In this study, we selected 18 layer hens with similar ovipositional rhythmicity (each of three birds were sacrificed for study per 4 h throughout 24 h); their transcriptomes were scanned to identify the circadian miRNAs and to explore regulatory mechanisms within the uterus of chickens. Results We identified six circadian miRNAs that are mainly associated with several biological processes including ion trans-membrane transportation, response to calcium ion, and enrichment of calcium signaling pathways. Verification of the experimental results revealed that miR-449c-5p exhibited a cosine expression pattern in the chicken uterus. Ca2+-transporting ATPase 4 (ATP2B4) in the plasma membrane is the predicted target gene of circadian miR-449c-5p and is highly enriched in the calcium signaling pathway. We speculated that clock-controlled miR-449c-5p regulated Ca2+ transportation during eggshell calcification in the chicken uterus by targeting ATP2B4. ATP2B4 mRNA and protein were rhythmically expressed in the chicken uterus, and dual-luciferase reporter gene assays confirmed that ATP2B4 was directly targeted by miR-449c-5p. The expression of miR-449c-5p showed an opposite trend to that of ATP2B4 within a 24 h cycle in the chicken uterus; it inhibited mRNA and protein expression of ATP2B4 in the uterine tubular gland cells. In addition, overexpression of ATP2B4 significantly decreased intracellular Ca2+ concentration (P < 0.05), while knockdown of ATP2B4 accelerated intracellular Ca2+ concentrations. We found similar results after ATP2B4 knockdown by miR-449c-5p. Taken together, these results indicate that ATP2B4 promotes uterine Ca2+ trans-epithelial transport. Conclusions Clock-controlled miR-449c-5p regulates Ca2+ transport in the chicken uterus by targeting ATP2B4 during eggshell calcification.

Aberrant Expression of Long Non Coding RNA HOTAIR and De-Regulation of the Paralogous 13 HOX Genes Are Strongly Associated with Aggressive Behavior of Gastro-Entero-Pancreatic Neuroendocrine Tumors

Gastro-entero-pancreatic neuroendocrine neoplasms (GEP-NENs) are rare diseases occurring in the gastrointestinal tract and pancreas. They are characterized by the loss of epithelial tubular gland elements, and by the increased expression of neuroendocrine markers. GEP-NENs are subdivided into two histo-pathological types, gastro-entero-pancreatic neuroendocrine tumors (GEP-NETs) and gastro-entero-pancreatic neuroendocrine carcinomas (GEP-NECs). According to WHO 2017 and 2019 classification criteria are graded and staged in four categories, NET-G1, NET-G2, NET-G3, and NEC-G3. The molecular characterization of these tumors can be fundamental for the identification of new diagnostic, prognostic and predictive biomarkers. The main purpose of this study was to analyze the expression of the paralogous 13 HOX genes, normally involved in embryogenic development and frequently deregulated in human cancers, and of the HOX regulating lncRNA HOTAIR in GEP-NENs. The expression of HOX genes is gradually lost in the transition from GEP NET G1 to NET/NEC G3 tumors, while HOTAIR expression, inversely correlated with HOX genes expression and weakly expressed in low-grade GEP NENs, becomes aberrant in NET G3 and NEC G3 categories. Our data highlights their potential role in the molecular stratification of GEP-NENs by suggesting new prognostic markers and potential therapeutic targets.

Effect of Fermented Kangkong Seed (Ipomoea reptans Poir.) on Characteristics and Histology of Female Quail Reproductive Organs

Quail has a high reproductive ability, which can reach 3 to 4 generations a year. The quail also have high eggs productivity and capable of producing 250-300 eggs/quail/year. Egg production is closely related to the reproductive system and feed. It is necessary to know the effect of feeding fermented kangkong on the characteristics and histology of the quail reproductive organs. The increase in livestock population should be balanced with an increase in the availability of feed. Kangkong seeds waste (Ipomoea reptans Poir.) is rejected seeds from seed factories that have the potential to become alternative feed ingredients because it contains about 13.46% protein. The purpose of this study was to determine the effect of fermented kangkong seed feed (Bikafer) on the characteristics of reproductive tract organs and histology of quail oviducts because feeds affect the development of the reproductive tract and quail egg production. This study used a completely randomized design method (CRD) with 5 treatments and 4 replications. The treatments were as follows: R0 = commercial feed, R1 = 0% (Bikafer), R2 = 7.5% (Bikafer), R3 = 15% (Bikafer) ,R4 = 22.5% (Bikafer). Data were analyzed using one-way ANOVA with a significance level of 95% (α = 0.05) and the Kruskal-Wallis test, while histological observations were described descriptively. The results showed that there were significant differences in the characteristics of the uterus, vaginal length, and isthmus weights. The histology of the quail reproductive tract developed well and showed the tunica mucosal tissue, serous tunica, muscular tunica, goblet cells, epithelium, and tubular gland cells.

Circadian miR-449c-5p regulates uterine Ca2+ transport during eggshell calcification in chickens

Background: miRNAs regulate circadian patterns by modulating animal biological clock. Clock genes exhibited a cosine expression pattern in the fallopian tube of chicken uterus in our previous study. Clock-controlled miRNAs are present in mammals and Drosophila; however, whether there are clock-controlled miRNAs in chicken uterus and, if so, how they regulate egg-laying rhythms are not clear. Here, we selected 18 layer hens with similar ovipositional rhythmicity (three birds were sacrificed for study per at 4 h intervals throughout 24 h); their transcriptomes were scanned to identify the circadian miRNAs and to explore regulatory mechanisms within the uterus of chickens. Results: We identified six circadian miRNAs mainly associated with several biological processes including ion trans-membrane transportation, response to calcium ion, and enrichment of calcium signaling pathways. Verification of experimental results revealed that miR-449c-5p exhibited a cosine expression pattern in chicken uterus. Ca2+-transporting ATPase 4 (ATP2B4) in the plasma membrane is the predicted target gene of circadian miR-449c-5p and is highly enriched in the calcium signaling pathway. We speculated that clock-controlled miR-449c-5p regulated Ca2+ transportation during eggshell calcification in chicken uterus by targeting ATP2B4. ATP2B4 mRNA and protein were rhythmically expressed in chicken uterus, and dual-luciferase reporter gene assays confirmed that ATP2B4 was directly targeted by miR-449c-5p. miR-449c-5p showed an opposite expression profile with ATP2B4 within a 24h cycle in chicken uterus; it inhibited mRNA and protein expressions of ATP2B4 in uterine tubular gland cells. Additionally, overexpression of ATP2B4 significantly decreased intracellular Ca2+ concentration (P < 0.05), while knockdown of ATP2B4 accelerated intracellular Ca2+ concentrations. Similar results were found after ATP2B4 knockdown by miR-449c-5p. These results indicated that ATP2B4 promoted uterine Ca2+ trans-epithelial transport. Conclusions: Clock-controlled miR-449c-5p regulates Ca2+ transport in chicken uterus by targeting ATP2B4 during eggshell calcification.

Circadian miR-449c-5p Regulates Uterine Ca2+ Transport During Eggshell Calcification in Chickens

Background: miRNAs regulate circadian patterns by modulating animal biological clock. Clock genes exhibited a cosine expression pattern in the fallopian tube of chicken uterus in our previous study. Clock-controlled miRNAs are present in mammals and Drosophila; however, whether there are clock-controlled miRNAs in chicken uterus and, if so, how they regulate egg-laying rhythms are not clear. Here, we selected 18 layer hens with similar ovipositional rhythmicity (three birds were sacrificed for study per at 4 h intervals throughout 24 h); their transcriptomes were scanned to identify the circadian miRNAs and to explore regulatory mechanisms within the uterus of chickens.Results: We identified six circadian miRNAs mainly associated with several biological processes including ion trans-membrane transportation, response to calcium ion, and enrichment of calcium signaling pathways. Verification of experimental results revealed that miR-449c-5p exhibited a cosine expression pattern in chicken uterus. Ca2+-transporting ATPase 4 (ATP2B4) in the plasma membrane is the predicted target gene of circadian miR-449c-5p and is highly enriched in the calcium signaling pathway. We speculated that clock-controlled miR-449c-5p regulated Ca2+ transportation during eggshell calcification in chicken uterus by targeting ATP2B4. ATP2B4 mRNA and protein were rhythmically expressed in chicken uterus, and dual-luciferase reporter gene assays confirmed that ATP2B4 was directly targeted by miR-449c-5p. miR-449c-5p showed an opposite expression profile with ATP2B4 within a 24h cycle in chicken uterus; it inhibited mRNA and protein expressions of ATP2B4 in uterine tubular gland cells. Additionally, overexpression of ATP2B4 significantly decreased intracellular Ca2+ concentration (P < 0.05), while knockdown of ATP2B4 accelerated intracellular Ca2+ concentrations. Similar results were found after ATP2B4 knockdown by miR-449c-5p. These results indicated that ATP2B4 promoted uterine Ca2+ trans-epithelial transport. Conclusions: Clock-controlled miR-449c-5p regulates Ca2+ transport in chicken uterus by targeting ATP2B4 during eggshell calcification.

A novel ability of endocytoscopy to diagnose histological grade of differentiation in T1 colorectal carcinomas

Background and study aims Endocytoscopic images closely resemble histopathology. We assessed whether endocytoscopy could be used to determine T1 colorectal cancer histological grade. Patients and methods Endocytoscopic images of 161 lesions were divided into three types: tubular gland lumens, unclear gland lumens, and fused gland formations on endocytoscopy (FGFE). We retrospectively compared endocytoscopic findings with histological grade in the resected specimen superficial layer, and examined the incidence of risk factors for lymph node metastasis. Results Of the 118 eligible lesions, the sensitivity, specificity, accuracy, negative predictive value, and positive likelihood ratio of tubular or unclear gland lumens to identify well-differentiated adenocarcinomas were 91.0 %, 93.1 %, 91.5 %, 77.1 %, and 13.20, respectively. To identify moderately differentiated adenocarcinomas for FGFE, these values were 93.1 %, 91.0 %, 91.5 %, 97.6 %, and 10.36, respectively. In the 35 lesions with FGFE, the rates of massive invasion, lymphovascular infiltration, and tumor budding were 97.1 %, 60.0 %, and 37.1 %, respectively. Conclusions Endocytoscopy could be used to diagnose T1 colorectal cancer histological grade, and FGFE was a marker for recommending surgery.

3. MORFOLOGI DAN KANDUNGAN KARBOHIDRAT KELENJAR LINGUALIS DAN SUBLINGUALIS ITIK (Anas plathyrinchos) (Morphology and Carbohydrates Content in Lingual and Sublingual Glands of Duck (Anas plathyrinchos))

This study was aimed to find out the morphology and carbohydrates content of lingual and sub-lingual glands of duck (Anas plathyrinchos). Sample used in this study was lingual and sublingual glands of a one year old female duck. Anatomy morphology and histology of lingual and sublingual glands were observed after processed into histological preparation and stained with hematoxylin-eosin (HE) then stained further with alcian blue (AB) pH 2.5 and periodic acid Schiff (PAS) to detect carbohydrate contents. The results showed that the lingual gland of duck cannot be observed due to its location below the epithelial layer of the tongue. While the sublingual gland of duck has an elongated shape and not granulated like typically glands, located at ventral tongue (dorsoventral lingual) between os mandibulae and covered with musculus digastricus venter oral. Staining with AB pH 2.5 and PAS showed that cytoplasm of secretory cell and secrete of lingual and sublingual glands contain acidic and neutral carbohydrates which indicated a positive reaction with intensity ranging from weak (+) to strong (+++). This study concluded that the lingual and sublingual gland found in female duck is a complex tubular gland with mucous secretory cells type. ____________________________________________________________________________________________________________________

Histological remarks of the oviduct and the oviducal gland of Sympterygia acuta Garman, 1877

The elasmobranchs constitute an important resource in Argentinian fishing and they show reproductive characteristics that make them susceptible to the pressure of fishing. In spite of the importance of the resource our knowledge about the species of the Southwestern Atlantic is scarce. In this work we study the microanatomy of the oviduct and the oviducal gland of Sympterygia acuta. The results show a very folded oviduct with mucous and ciliated cells. The oviducal gland depicts the same zonation as other batoids, showing the typical four regions (club, papillary, baffle and terminal). The epithelium lining the glandular lumen is simple columnar with ciliated and glandular cells. The four zones show simple or ramified tubular gland the secretions of which constitute the egg's envelopes. The club and baffle zones are similar to those present in other species. The papillary one depicts a different pattern of secretions that other Rajiformes and the last zone of the gland is characterized by mixed adenomers. The information here exposed constitutes the first report on the microanatomy of the genital tract of S. acuta.