Duct Formation Research Articles

Herlyn-Werner-Wunderlich Syndrome: A Case Report on a Congenital Uterine Anomaly With Literature Review

During fetal development, abnormalities during Mullerian duct formation can lead to varied types of uterine, cervical, or vaginal anomalies. Herlyn-Werner-Wunderlich Syndrome (HWWS) is a rare, congenital Mullerian duct anomaly characterized by the triad of uterine didelphys, obstructed hemi-vagina, and ipsilateral renal agenesis. If the diagnosis is delayed or missed, permanent sequalae such as chronic pain, infection, and infertility can result. This is a case of a 16-year-old woman with heavy vaginal bleeding and menstrual cramping in which sonography was used in the diagnosis of HWWS. Sonography plays a vital role in this diagnosis, as it is the most common initial imaging examination. For this reason, it is important that the characteristics of uterine anomalies, like HWWS, be recognized with sonography. It is important that this diagnosis is made early and treatment can be initiated to prevent irreversible complications.

Population differentiation in climate sensitivity of resin duct formation during growth resumption in Pinus pinaster

To counteract the effects of herbivores and pathogens, conifers have developed a sophisticated resin-based defensive system. Since defences are costly, trees must continuously accommodate defensive investment throughout plastic responses to environmental stimuli. However, the extent of such responses can differ at the intra-specific level (i.e. genetic variation in plasticity). Here we examined whether and to what extent year-to-year climate fluctuations, an important source of environmental heterogeneity during the trees' life, drive plasticity in defensive allocation of a widespread pine species. Specifically, we quantified interannual variation in resin duct production along a 31-year-period in 174 Pinus pinaster trees of nine range-wide populations grown in two common gardens in Central Spain. We aimed to explore (i) patterns of interannual variation (i.e., temporal plasticity) in resin duct production among populations and sites, (ii) whether such patterns are linked to plastic responses to interannual variation in climate conditions (i.e., climatic plasticity), and (iii) whether plastic responses to climate differ among populations (i.e., genetic variation in plasticity) and sites. We found large interannual plasticity in resin duct production (22.8 % of total variance), with temporal patterns differing among sites and populations. Climate conditions during the early growth period significantly affected the annual differentiation of resin ducts. Particularly, April precipitation had a positive overall effect on resin duct production. Inversely, warmer conditions in April had a negative effect but only in certain populations, which demonstrates genetic variation in climate sensitivity of resin duct formation. Despite significant effects of certain climate variables on annual resin duct production, climate only accounted for a small proportion of the total interannual variation (up to 3.8 % of interannual variation explained by climate factors). This suggests that alternative factors such as trade-offs with growth and temporal variation in biotic and non-climatic abiotic conditions likely contribute to explain interannual fluctuations in defensive investment.

Mechanistic Drivers of Müllerian Duct Development and Differentiation Into the Oviduct.

The conduits of life; the animal oviducts and human fallopian tubes are of paramount importance for reproduction in amniotes. They connect the ovary with the uterus and are essential for fertility. They provide the appropriate environment for gamete maintenance, fertilization and preimplantation embryonic development. However, serious pathologies, such as ectopic pregnancy, malignancy and severe infections, occur in the oviducts. They can have drastic effects on fertility, and some are life-threatening. Despite the crucial importance of the oviducts in life, relatively little is known about the molecular drivers underpinning the embryonic development of their precursor structures, the Müllerian ducts, and their successive differentiation and maturation. The Müllerian ducts are simple rudimentary tubes comprised of an epithelial lumen surrounded by a mesenchymal layer. They differentiate into most of the adult female reproductive tract (FRT). The earliest sign of Müllerian duct formation is the thickening of the anterior mesonephric coelomic epithelium to form a placode of two distinct progenitor cells. It is proposed that one subset of progenitor cells undergoes partial epithelial-mesenchymal transition (pEMT), differentiating into immature Müllerian luminal cells, and another subset undergoes complete EMT to become Müllerian mesenchymal cells. These cells invaginate and proliferate forming the Müllerian ducts. Subsequently, pEMT would be reversed to generate differentiated epithelial cells lining the fully formed Müllerian lumen. The anterior Müllerian epithelial cells further specialize into the oviduct epithelial subtypes. This review highlights the key established molecular and genetic determinants of the processes involved in Müllerian duct development and the differentiation of its upper segment into oviducts. Furthermore, an extensive genome-wide survey of mouse knockout lines displaying Müllerian or oviduct phenotypes was undertaken. In addition to widely established genetic determinants of Müllerian duct development, our search has identified surprising associations between loss-of-function of several genes and high-penetrance abnormalities in the Müllerian duct and/or oviducts. Remarkably, these associations have not been investigated in any detail. Finally, we discuss future directions for research on Müllerian duct development and oviducts.

Development of Testis Cords and the Formation of Efferent Ducts in Xenopus laevis: Differences and Similarities with Other Vertebrates

The knowledge of testis development in amphibians relative to amniotes remains limited. Here, we used Xenopus laevis to investigate the process of testis cord development. Morphological observations revealed the presence of segmental gonomeres consisting of medullary knots in male gonads at stages 52–53, with no distinct gonomeres in female gonads. Further observations showed that cell proliferation occurs at specific sites along the anterior-posterior axis of the future testis at stage 50, which contributes to the formation of medullary knots. At stage 53, adjacent gonomeres become close to each other, resulting in fusion; then (pre-)Sertoli cells aggregate and form primitive testis cords, which ultimately become testis cords when germ cells are present inside. The process of testis cord formation in X. laevis appears to be more complex than in amniotes. Strikingly, steroidogenic cells appear earlier than (pre-)Sertoli cells in differentiating testes of X. laevis, which differs from earlier differentiation of (pre-)Sertoli cells in amniotes. Importantly, we found that the mesonephros is connected to the testis gonomere at a specific site at early larval stages and that these connections become efferent ducts after metamorphosis, which challenges the previous concept that the mesonephric side and the gonadal side initially develop in isolation and then connect to each other in amphibians and amniotes.

GATA6 modulates the ductular reaction to bile duct ligation.

GATA6, a transcription factor expressed in cholangiocytes, has been implicated in the response to liver injury. In biliary atresia, a disease characterized by extrahepatic bile duct obstruction, liver expression of GATA6 increases with pathological bile duct expansion and decreases after successful Kasai portoenterostomy. The aim of this study was to garner genetic evidence that GATA6 is involved in ductular formation/expansion. The murine Gata6 gene was conditionally deleted using Alb-cre, a transgene expressed in hepatoblasts (the precursors of hepatocytes and cholangiocytes) and mature hepatocytes. Bile duct ligation (BDL) was used to model biliary obstruction. Alb-Cre;Gata6flox/flox mice were viable and fertile. Cre-mediated recombination of Gata6 in hepatocytes had little impact on cellular structure or function. GATA6 immunoreactivity was retained in a majority of biliary epithelial cells in adult Alb-Cre;Gata6flox/flox mice, implying that surviving cholangiocytes were derived from hepatoblasts that had escaped biallelic Cre-mediated recombination. Although GATA6 immunoreactivity was preserved in cholangiocytes, Alb-cre;Gata6flox/flox mice had a demonstrable biliary phenotype. A neutrophil-rich infiltrate surrounded newly formed bile ducts in neonatal Alb-Cre;Gata6flox/flox mice. Foci of fibrosis/necrosis, presumed to reflect patchy defects in bile duct formation, were observed in the livers of 37% of adult Alb-cre;Gata6flox/flox mice and 0% of controls (p < 0.05). Most notably, Alb-cre;Gata6flox/flox mice had an altered response to BDL manifest as reduced survival, impaired bile ductule proliferation, increased parenchymal necrosis, reduced fibrosis, and enhanced macrophage accumulation in the portal space. GATA6 orchestrates intrahepatic biliary remodeling and mitigates liver injury following extrahepatic bile duct obstruction.

TRANSFORMATION OF THE HUMAN PROSTATE GLAND IN INTERMEDIATE AND LATE FETAL PERIODS OF DEVELOPMENT

The purpose of the study was to determinate the morphometric parameters of epithelial cords, prostatic ducts, acini of the human prostate gland and their lining epithelium, the shape of prostate glands, and muscle tissue in fetal periods of development. Material and Method: The study was performed on 19 prostate glands of male sex fetuses of intermediate and late fetal periods. We used the morphometric method of verification of apoptotic cells on stained histoligical preps using the criteria of apoptosis. Morphometric research included measurements of acini and acinar lumen areas, epithelium height, assessment of shape factors. Results: In the late fetal period compared to the intermediate fetal period the acini area was not increased in 1.1 times (p =0,004), the area of acinar lumens increases by 3.8 times (p=0.0005). Changes in the shape of acinar lumens were detected. Conclusion: An insignificant increase of the specific area of the glandular parenchyma in the organ of transformation of the developing glands occurs mainly due to their canalization. The formation of prostatic ducts was not followed by the increase in their total area and was the result of differentiation epithelial cells and apoptosis. Transformation of prostate glands in the prenatal period occurs in a certain sequence: the formation of epithelial buds, their canalization by apoptosis with formation of epithelial ducts and prostatic ducts, and formation of end pieces of prostate glands from these ducts. Apoptotic bodies were removed from the lumen of the prostatic ducts by extrusion. The design and orientation of the smooth muscle bundles around prostatic ducts and acini do not ensure the evacuation of content of the prostate glands in the fetal period.

Neuropilin-1 directs PDGFRα-entry into lung fibroblasts and signaling from very early endosomes.

Platelet-derived growth factor receptor-α (PDGFRα) is absolutely required for the development of secondary pulmonary alveolar septa. Our earlier observations indicated that PDGFRα resides intracellularly as well as on the plasma membrane of murine lung fibroblasts (LF). We have examined how neuropilin-1 (Nrp1), a surface receptor without kinase activity, regulates the intracellular trafficking of PDGFRα in LF obtained from mice, some bearing a targeted deletion of Nrp1 in myofibroblasts. Using the proximity ligation assay, we observed that PDGFRα and Nrp1 colocalized in both early antigen-1 (EEA1) containing sorting endosomes and with adaptor protein containing a pleckstrin homology domain and a phosphotyrosine-binding domain-1 (APPL1) in very early endosomes (VEE). These findings were confirmed using live-cell imaging, which demonstrated that recently internalized PDGFRα was observed in Rab5-containing vesicles residing within 100 nm of the plasma membrane. Nrp1 deletion reduced the phosphorylation of Akt (protein kinase B), the major downstream target of PDGFRα, and limited accumulation of inositol-3 phosphates in APPL1-containing endosomes after exposure to PDGFA. PDGFRα co-immunoprecipitated with APPL1, indicating that PDGFRα enters VEE. Targeted deletion of Nrp1 or APPL1-depletion in control LF reduced the activity of an Akt1 biosensor following stimulation with PDGFA. Our findings demonstrate that Nrp1 enhances the entry of PDGFRα into APPL1 containing VEE and that APPL1 enhances PDGFRα signaling. Therefore, Nrp1 promotes endosomal signaling by PDGFRα offering a potential mechanism to explain our prior observation that Nrp1 supports the formation of alveolar ducts and alveoli during secondary septation in mice.

Effects of Rising Temperature on Microwave Communications in Ducting Environment over the Southern Region of Pakistan in the Northern Arabian Sea

The propagation of microwave (MW) of frequencies above 300 MHz is affected by the existence and properties of the atmospheric duct. Atmospheric ducts exist in many areas of the world ocean, including the Arabian Sea. Located in the Hadley Cell and monsoon region, different seasons bring air masses of different properties into the area under investigation, which has a significant impact on the formation and strength of the atmospheric duct. In this paper, we have done the modeling to analyze the patterns of electromagnetic ducting, which is significant in the southern region of Pakistan in the Arabian Sea. To analyze the real-time scenario, data were collected from three different areas off the southern coast of Pakistan in the northern Arabian Sea to observe the electromagnetic wave's effect on the evaporation ducts. Our analysis results reveal that rising temperature plays a significant role where ducts occur above 30% in the summer months and less than 7% in the spring, autumn, and winter months. It is due to an increase in temperature, especially in summer and autumn months, where humidity gradients play an essential role in creating a higher frequency of duct. The same observations were simulated to view the time analysis of pressure, humidity, and potential temperature in this region, depending upon the refractive index.

Anatomical study of ethylene induced gum duct formation and scientific gum tapping technique in gum Karaya (Sterculiaurens Roxb.)

Gums and resins occupy a prime place among Non-Wood Forest Produce (NWFP/NTFP) and are known to mankind since time immemorial. Gums are metabolic by-products of plant tissues either in normal course or often as a result of disease or injury to the bark or wood of certain plants and it cannot be re-enter with plant system. Gum Karaya (Sterculia urens Roxb.) is a dry deciduous tree belonging to the family Sterculiaceae distributed throughout India and Chhattisgarh. The impact of gum enhance ethephon was found significantly superior, regarding the production of biopolymers. The biological (anatomical), studies were done via taking the sample of soft (sapwood) after injecting the gum enhancer ethephon at different time intervals. It can observed that the application of ethephon enhance the process of gummosis due to formation of gum duct. The gum duct formation was observed in histological analysis of bark section within 2 hrs of ethephon treatment in gum Karaya. The histological changes indicated that the gum ducts lysigenously are present in the pith and cortex of the young stem of Sterculia urens but absent in the xylem. The commercial tapping of Karaya is done by blazing, peeling, or by making deep cuts at the base of the bole using an axe or a cycle. These methods often lead to the death of the tapped trees. On account of crude tapping methods and over exploitation the population of karaya trees has markedly declined. The harvesting method currently used are traditional and injurious due to which often obtained inferior quality of products. Hence, the study was undertaken in ICAR Network Project to develop the scientific tapping technique for sustainable harvesting in major gum producing tree of Chhattisgarh state to enhance the livelihood of the rural areas as well as to protect the plant and generate the revenue of the government. The chemical tapping method using ethephon and IAA injected by battery operated drill machine. However, temperature and relative humidity also play significant role in gum exudation. The ethephon @ 3.9% in 4 ml in two consecutive doses in 45-60 days intervals at high temperature in month of April to June was found significantly effective for maximum gum production during both the year 2015 and 2016. The physiochemical properties of exudated gum were investigated and gum was found to be mild acidic, least soluble in cold water but absorb water and swell, soluble in hot water but insoluble in organic solvent.

Trampling as a major ecological factor affecting the radial growth and wood anatomy of Scots pine (Pinus sylvestris L.) roots on a hiking trail

The impact of hiking in forested areas is one of the main factors affecting the condition of tree growth along hiking trails. Trampling causes common exposure of roots and quantification of the human impact on root radial growth and wood anatomy requires careful assessment. To accurately identify the radial growth changes in Pinus sylvestris roots, we conducted a stepwise cross-dating using trampled roots from a hiking trail and reference roots. Such approach was previously not applied in a lowland area. In addition, we investigated the factors that influence the root-stem radial growth coherency, including soil properties, root and trail morphology. Changes in radial growth and wood anatomy were examined in three parts of the root system: buried, transition and trampling zone. The radial growth for each root zone was compared with corresponding stem and reference root and stem chronology. In total, we investigated 204 roots and 97 tree stems for the common period 1970–2015. Missing rings were found to be a common phenomenon in all root zones, except in the exposed lateral roots in the trampling zone. The highest number of wedging and missing rings was observed in the trampling and transition zones, respectively. The total number of wedging rings increased with an increasing distance from the stem. The events of root exposure in the trampling zone were highly coupled with the formation of scars (r = 0.51, p < 0.001) and pronounced resin ducts (PRDs) (r = 0.52, p < 0.001). The majority of the wood anatomical changes (i.e., 84% of scars and 85% of PRDs) were identified in the trampling zone. The highest degree of correlation between raw stem chronology and raw root chronology was found for the exposed trampled roots (r = 0.69, p < 0.001). In the trampling zone, the root-stem radial growth coherency was determined primarily by root type (77.3%) and root age (17.4%), whereas in the buried zone, by the soil organic matter content (55.4%) and soil compaction (39.7%). It has been proved that the record of radial growth and wood anatomy changes in Scots pine roots serve as a valuable ecological archive of trampling impact with high temporal resolution.

The zonula occludens protein family regulates the hepatic barrier system in the murine liver

The hepatic barrier is indispensable for the physiological functions of the liver and is impaired under various pathological conditions. Tight junctions reportedly play a central role in hepatic barrier regulation; however, there is limited direct evidence supporting this observation, with few in vivo models or confirmations of the implicated molecular mechanisms presented to date. We inactivated the tight junction component gene, Tjp2/ZO-2, and the related molecule, Tjp1/ZO-1, in mouse livers. In humans, TJP2/ZO-2 mutations have been implicated in the development of human progressive familial intrahepatic cholestasis 4 (PFIC4). The mice deficient in either ZO-1 or ZO-2 in the liver did not exhibit major abnormalities. However, the ablation of both molecules impaired the molecular architecture as well as the structure and function of hepatocyte tight junctions, which disrupted the hepatic barrier and was lethal to the mice by 6 weeks of age. In mutant mice, bile canaliculus formation and cellular polarity were compromised; also, transporter expression and localization were deregulated. Moreover, typical hepatic zonation and bile duct formation were inhibited, and sinusoidal vessels were disorganized. These findings clarify the role of tight junctions and polarity in the hepatic barrier as well as the effect that their disruption has on liver tissue. The observations also suggest that liver-specific ZO-1−/− and ZO-2−/− mice could be used as models for PFIC4, and this will provide new insights into liver pathophysiology and clinical applications.

Transcriptional landscape of the embryonic chicken M\xfcllerian duct

BackgroundMüllerian ducts are paired embryonic tubes that give rise to the female reproductive tract in vertebrates. Many disorders of female reproduction can be attributed to anomalies of Müllerian duct development. However, the molecular genetics of Müllerian duct formation is poorly understood and most disorders of duct development have unknown etiology. In this study, we describe for the first time the transcriptional landscape of the embryonic Müllerian duct, using the chicken embryo as a model system. RNA sequencing was conducted at 1 day intervals during duct formation to identify developmentally-regulated genes, validated by in situ hybridization.ResultsThis analysis detected hundreds of genes specifically up-regulated during duct morphogenesis. Gene ontology and pathway analysis revealed enrichment for developmental pathways associated with cell adhesion, cell migration and proliferation, ERK and WNT signaling, and, interestingly, axonal guidance. The latter included factors linked to neuronal cell migration or axonal outgrowth, such as Ephrin B2, netrin receptor, SLIT1 and class A semaphorins. A number of transcriptional modules were identified that centred around key hub genes specifying matrix-associated signaling factors; SPOCK1, HTRA3 and ADGRD1. Several novel regulators of the WNT and TFG-β signaling pathway were identified in Müllerian ducts, including APCDD1 and DKK1, BMP3 and TGFBI. A number of novel transcription factors were also identified, including OSR1, FOXE1, PRICKLE1, TSHZ3 and SMARCA2. In addition, over 100 long non-coding RNAs (lncRNAs) were expressed during duct formation.ConclusionsThis study provides a rich resource of new candidate genes for Müllerian duct development and its disorders. It also sheds light on the molecular pathways engaged during tubulogenesis, a fundamental process in embryonic development.

S2531 A Rare Cause of Cholestasis: Secondary Sclerosing Cholangitis in a Critically Ill Patient

INTRODUCTION: Secondary sclerosing cholangitis in critically ill patient (SSC-CIP) is a newly diagnosed entity of severe biliary disease which can rapidly progress to liver cirrhosis. The mechanism leading to this form of cholangitis with stricture formation and complete obliteration of bile ducts is unknown (1). CASE DESCRIPTION/METHODS: 47 year old woman with history of roux en y gastric bypass was admitted for status epilepticus, pneumonia and shock requiring vasopressors and ventilator support for over a month in ICU. Patient’s LFTs at discharge were normal. Patient was readmitted few weeks later with a seizure and found to have abnormal LFTs (ALP 2500, GGT 4000, direct bilirubin 3.2, ALT 80, and AST 75, lipase 80) which persisted over the course of next ten months. The negative work up included HAV, HBV, HCV, HIV, ANA, AMA, ASMA, Immunoglobulins, Anti LKM, ANCA, Anti SP 100, Anti GP 210, ceruloplasmin, alpha 1 anti-trypsin, TTG IgG, TSH, AFP, CA19-9 and leukemia/lymphoma markers. Her ultrasound and CT scan was unremarkable. Patient was unable to get a MRCP due to a vagal nerve stimulator and ERCP was not done due to difficult anatomy. She underwent a IR guided cholangiogram; subtle stricturing and mild dilation of left intrahepatic ducts seen, followed by a liver biopsy. Biopsy showed large bile duct obstruction with secondary sclerosing cholangitis due to ischemic cholangiopathy with advanced biliary fibrosis. Patient was started on ursodiol with improvement of LFTs however she got readmitted few days later with hemorrhagic shock due to delayed bleed from the liver biopsy site with sepsis and multiorgan failure resulting in death. DISCUSSION: SSC-CIP can be caused by events such as prolonged hypotension and respiratory insufficiency that lead to severe irreversible injury of ducts (2). The trigger event of ischemic cholangoipathy in our patient was likely her initial critical illness (hypotension and respiratory failure) resulting in cholestasis. This variant SSC-CIP is a relatively new disease that is an under recognized cause of cholestasis and should be suspected as a differential in patients with new cholestasis after a critical illness.

Development of Ectopic Livers by Hepatocyte Transplantation Into Swine Lymph Nodes.

Orthotopic liver transplantation continues to be the only effective therapy for patients with end‐stage liver disease. Unfortunately, many of these patients are not considered transplant candidates, lacking effective therapeutic options that would address both the irreversible progression of their hepatic failure and the control of their portal hypertension. In this prospective study, a swine model was exploited to induce subacute liver failure. Autologous hepatocytes, isolated from the left hepatic lobe, were transplanted into the mesenteric lymph nodes (LNs) by direct cell injection. At 30‐60 days after transplantation, hepatocyte engraftment in LNs was successfully identified in all transplanted animals with the degree of ectopic liver mass detected being proportional to the induced native liver injury. These ectopic livers developed within the LNs showed remarkable histologic features of swine hepatic lobules, including the formation of sinusoids and bile ducts. On the basis of our previous tyrosinemic mouse model and the present pig models of induced subacute liver failure, the generation of auxiliary liver tissue using the LNs as hepatocyte engraftment sites represents a potential therapeutic approach to supplement declining hepatic function in the treatment of liver disease.

POLYMORPHOUS LOW-GRADE ADENOCARCINOMA IN THE SUBMANDIBULAR REGION: A CASE REPORT

The objective of the report is to present a case of polymorphic adenocarcinoma that presents with aggressive local behavior and bone invasion. The patient was treated in May 2017 with swelling in the right mandible of hardened consistency, asymptomatic, and 1-year evolution. The panoramic radiograph showed an osteolytic radiolucent lesion with poorly defined margins and cortical rupture. In the incisional biopsy, glandular origin’s malignant neoplastic cells were found, in a predominantly solid invasion pattern and ducts formation, justifying the histopathologic diagnosis of adenocarcinoma not specified. Afterward, the patient was referred to oncology, where he was submitted to a glosso-mandibulectomy, considering the procedure performed (removal of the whole jaw) with neck dissection in June 2017. The surgical specimen’s diagnosis showed polymorphous adenocarcinoma, presenting bone invasion without neural and vascular invasion. The patient has been under care for 2 years and shows no sign of recurrence of the neoplasia.

Occurrence of metastatic eccrine porocarcinoma in an immunosuppressed patient

BackgroundEccrine porocarcinoma is a rare malignancy accounting for 0.005 to 0.01% of all cutaneous tumors. However, its etiology is not well established. Herein we are reporting the occurrence of metastatic porocarcinoma of the back in a patient with a history of multiple cutaneous malignancies and chronic immunosuppression.Case presentationA 79-year-old male with a history of long-term immunosuppressive therapy for left lung transplantation and multiple cutaneous malignancies including melanoma, presented with an enlargening plaque on the right upper back. Biopsy demonstrated an infiltrative epithelial tumor with aggressive histology and ductal formation. Immunohistochemical stains for p63, CK-5/6, and CEA were diffusely positive, while CK-7 and CK-20 were negative. A diagnosis of eccrine porocarcinoma was made. He underwent complete wide excision with negative margins of the right upper back in January 2018. In November 2018, he was found to have a right posterior shoulder mass, positive right axillary lymph nodes, superficial right upper back soft tissue masses and lymphangitic carcinomatosis involving the left lung. He was transitioned to comfort care and subsequently passed from metastatic disease in December 2018.ConclusionThe occurrence of eccrine porocarcinoma in a patient on long term immunosuppressive therapy for lung transplantation suggests a possible role for chronic immunosuppression in the induction of a subset of eccrine porocarcinomas, as in other cutaneous malignancies.

A CASE OF FUNCTIONING NON COMMUNICATING RUDIMENTARY HORN WITH UNICORNUATE UTERUS: AN UNUSUAL CAUSE OF SECONDARY DYSMENORRHEA IN A PERIMENOPAUSAL WOMAN.

Mullerian duct malformations represent a miscellaneous group of congenital anomalies that result from the arrested development, abnormal formation or incomplete fusion of the paramesonephric ducts. Congenital Mullerian abnormalities usually present at adolescent age with menstrual irregulari¬ty, dysmenorrhea, infertility and recurrent pregnancy loss. The Unicornuate uterus is a rare uterine malformation which usually features a rudimentary accessory horn with or without functioning endometrium. We are presenting a case of perimenopausal multiparous patient who had history of secondary dysmenorrhea diagnosed with endometrioma on imaging studies, but on laparotomy incidentally found to have Unicornuate uterus with functioning noncommunicating rudimentary horn leading to hematometra and hematosalpinx. The objective of this clinical case report is to highlight this rare Mullerian malformation as a differential diagnosis of secondary dysmenorrhea, chronic pelvic pain or adnexal mass in perimenopausal age group females.

Novel ex vivo disease model for extramammary Paget’s disease using the cancer tissue-originated spheroid method

BackgroundExtramammary Paget’s disease (EMPD) is a rare skin cancer that frequently occurs in the anogenital region in the elderly. Prognosis in patients with metastatic EMPD is poor as EMPD treatment has advanced little in recent years, primarily because no EMPD cell line has been established. ObjectiveWe aimed to establish an ex vivo EMPD disease model using the cancer tissue-originated spheroid (CTOS) method, which is used to prepare and culture primary cancer cells while maintaining cell–cell contact. MethodsThirteen samples from 12 EMPD patients were obtained. CTOSs were prepared and cultured using CTOS method. Histopathological examination of the CTOSs was performed. We investigated optimum medium conditions and effects of growth factors for CTOS growth. Chemo-sensitivity assays were conducted. ResultsCTOSs were successfully prepared from 3 primary lesions and 2 metastatic lymph nodes. Of these, 2 CTOSs (EMPD-3 and EMPD-4) could be maintained and passaged long term ex vivo. Following transplantation of CTOSs to NOD/Scid mice, CTOS-derived xenotumors exhibited ductal formation, indicating that CTOSs retained the original tumor characteristics. Chemo-sensitivity assays revealed that docetaxel significantly inhibited EMPD-3 growth in a dose-dependent manner, whereas EMPD-4 was not clearly inhibited. These findings indicate the heterogeneity of EMPD and potential use of chemosensitivity assays with patient-derived CTOS to select the most effective drugs for each patient. ConclusionTo our knowledge, this study represents the first establishment of an ex vivo-EMPD disease model involving conventional cell lines. EMPD CTOSs might be useful for developing new therapeutic strategies.

Astragaloside IV regulates the HIF/VEGF/Notch signaling pathway through miRNA-210 to promote angiogenesis after ischemic stroke.

Astragaloside IV (AS-IV) is one of the main active ingredients of Astragalusmembranaceus. Studies have shown that AS-IV stimulates angiogenesis, including cell proliferation, migration, and neovascularization. However, the relevant mechanism remains unclear. This study aims to investigate whether AS-IV activates the HIF/VEGF/Notch signaling pathway through miRNA-210 to promote angiogenesisafter ischemic stroke. The present study established a rat model of middle cerebral artery occlusion (MCAO) and cultured human umbilical vein endothelial cells (HUVECs) under hypoxic conditions in vitro to investigate the role of AS-IV in promoting angiogenesis and reveal its underlying mechanism. Through in vivo studies, the area of cerebral infarction was determined by 2,3,5-triPhenyltetrazolium chloride (TTC) staining. Immunofluorescence staining and RT-qPCR were used to detect the expression changes of miRNA-210 and ephrinA3 in the ischemic cortex after ischemia. Through in vitro studies, cell proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Subsequently, angiogenesis experiments were performed to observe the angiogenic ability. Results revealed that AS-IV significantly reduced infarct size, promoted cell proliferation and ductal formation, and inhibited the expression of the target gene ephrinA3 by increasing the expression of miRNA-210 and inducing activation of the HIF-VEGF-Notch signaling pathway. AS-IV promotes cerebral protection following angiogenesis and ischemic brain injury. The specific mechanism was activating the HIF/VEGF/Notch signaling pathway via miRNA-210.

Monitoring and maintaining the differentiation status of human ipsc derived kidney organoids

Background & Aim Human induced pluripotent stem cell (iPSC) derived kidney organoids resemble first trimester fetal kidney and offer a great model for studying kidney development and disease. During early renal development, the inductive interaction between two intermediate mesoderm (IM)-derived progenitor cell populations, the ureteric bud and the kidney mesenchyme, promotes nephron and collecting duct formation. Protocols are developed to in vitro differentiate human iPSC into ureteric epithelium and metanephric mesenchyme simultaneously, and then aggregate them to generate a complex multicellular kidney organoids. However, this is a complex procedure with unpredictable outcomes. Furthermore, after 25 days of culture, kidney organoids start to degenerate and form fibrotic and cartilage tissue. We investigated how kidney organoid differentiation can be monitored and how the survival of kidney organoids can be prolonged. Methods, Results & Conclusion We treated five human iPSC with CHIR99021 and followed by FGF9 to induce IM for 7 days. These IM like cells were formed into cell pellets and transferred to air-liquid interface for a further 18 up to 32 days. Successfully differentiated organoids demonstrate glomerular and tubular structures in H&E section, while failed differentiation results in amorphous fibrotic tissue. The same iPSC line had variable results among batch to batch. Along the differentiation process, LDH activity was measured to monitor organoid metabolism, which reached a peak on days 7∼9. LDH peaks higher than 50 milliunits/ml indicated excessive necrosis and resulted in abnormal organoid formation. We observed that iPSC with elevated OCT4 expression showed induced capacity to differentiate into kidney organoids. Furthermore high TGFβ expression at day 7 was associated with fibrosis in the kidney organoids. As shortage of oxygen supply to kidney organoids due to lack of functional vasculature limits their long term stability, we split the organoids on day 8 and day 21 and placed the organoids on a shaker on day 21 to allow for better oxygen supply. Using this method, it was possible to preserve the kidney organoids for 2 additional weeks with maintenance of glomerular and tubular structures. Taken together, these data will help us to monitor the differentiation and prolong the culture time of kidney organoids.