The contribution of TEM to solving issues in the oogenesis of lower metazoans: a comparison between Acoela and rhabditophoran Platyhelminthes

The chapter reports data on the oogenesis and eggshell formation in some Acoela obtained by our group over the years by means of transmission electron microscopy (TEM) and cytochemical techniques. In this review we show how TEM and appropriate cytochemical methods have been helpful in clarifying issues related to female germ cell differentiation and more in general to the cellular aspects of the reproductive biology in these lower metazoans. The ultrastructural findings have been compared with those from other Acoelomorpha (Acoela + Nemertodermatida) and rhabditophoran Platyhelminthe

Complete intraperitoneal displacement of a double J stent: a first case.

OBJECTIVES: Ureteral double-J stents are known to migrate proximally and distally within the urinary tract, while perforation and stent displacement are uncommon. Possible mechanisms of displacement are either original malpositioning with ureteral perforation or subsequent fistula and erosion of the excretory system, due to infection or long permanence of the device. We present the unique case of complete intraperitoneal stent migration in a 59-year-old caucasian male without evidence of urinary fistula at the moment of diagnosis, so far an unreported complication. MATERIALS AND METHODS: Eight months after the placement of a double-J stent for lower right ureteral stricture at a district hospital, the patient came at our observation for urosepsis and hydro-uretero-nephrosis. A CT scan demonstrated intraperitoneal migration of the stent outside the urinary tract. Cystoscopy failed to visualize the lower extremity of the stent, a percutaneous nephrostomy was placed to drain the urinary system and the stent was removed through a small abdominal incision on the right lower quadrant. RESULTS: In our case we presume that during the positioning manoeuvre the guide wire perforated simultaneously the lower ureteral wall and the pelvic peritoneum, and that once the upper end of the stent was coiled, the lower extremity was also attracted intraperitoneally. The lack of pain due to the spinal lesion concurred to this unusual complication. CONCLUSIONS: We must be aware that ureteral double J stents may be found displaced even inside the peritoneal cavity, and that the use of retrograde pyelography during placement is of paramount importance to exclude misplacement of an apparently normally coiled upper extremity of the stent

Innovative Interprofessional Collaboration: Addressing the Need for Education for Health Center Workers around Patients\u27 Sexual Orientation and Gender Identity (SOGI)

BACKGROUND: University of Rhode Island (URI) students in a graduate-level course, Interdisciplinary Teamwork in Health and Human Services, collaborated with a local health provider, Wood River Health Services (WRHS), to create training materials for its staff on the topic of sexual orientation/gender identity (SOGI). WRHS is a federally-funded, comprehensive community health center located in a rural area of Rhode Island. It provides medical, dental, and mental health services. WRHS had been advised to enhance its attention to issues related to SOGI. OBJECTIVES The aims for this project were: 1. Assess needs and priorities of WRHS staff around knowledge and attitudes about SOGI and health care; 2. Create a training tool for staf fto enable them to facilitate patient comfort in disclosing SOGI-related information that would assist them in addressing individual needs of patients; 3. Provide students an opportunity to effectively learn and practice interprofessionally. METHODS The students--representing three different health fields (Occupational Therapy, Human Development and Family Studies, and Pharmacy)--researched SOGI issues related to their fields. Students met with WRHS staff to develop a clear understanding of the center’s barriers to adequately provide care to LGBTQ+ patients. They conducted a survey to assess staff attitudes and knowledge about SOGI terminology, health disparities, and the importance of a welcoming environment that informed the development of a training presentation. Staf can complete it at their convenience, and it can be used as part of onboarding new staff. Retrospective pre/post-tests were administered to students and WRHS staff. CONCLUSION Informational presentations are a useful tool for agencies looking to build capacity and comfort among staff around SOGI. Additionally, providing students with opportunities to learn with, from, and about each other is a critical component of health professions education.https://jdc.jefferson.edu/sexandgenderhealth/1020/thumbnail.jp

Unexpected thymoma in a challenging case of hyperparathyroidism

We report the case of a woman with primary hyperparathyroidism suspected of mediastinal ectopic parathyroid adenoma revealed to be a thymoma. Our aim was to focus on some possible criticisms in distinguishing between ectopic parathyroid and thymus

Connexin 26 Expression in Mammalian Cardiomyocytes

Connexins are a family of membrane-spanning proteins named according to their molecular weight. They are known to form membrane channels mediating cell-cell communication, which play an essential role in the propagation of electrical activity in the heart. Cx26 has been described in a number of tissues but not in the heart, and its mutations are frequently associated with deafness and skin diseases. The aim of this study was to assess the possible Cx26 expression in heart tissues of different mammalian species and to demonstrate its localization at level of cardiomyocytes. Samples of pig, human and rat heart and H9c2 cells were used for our research. Immunohistochemical and molecular biology techniques were employed to test the expression of Cx26. Interestingly, this connexin was found in cardiomyocytes, at level of clusters scattered over the cell cytoplasm but not at level of the intercalated discs where the other cardiac connexins are usually located. Furthermore, the expression of Cx26 in H9c2 myoblast cells increased when they were differentiated into cardiac-like phenotype. To our knowledge, the expression of Cx26 in pig, human and rat has been demonstrated for the first time in the present paper

Inflammatory role of extracellular sphingolipids in Cystic Fibrosis

Ceramide is emerging as one of the players of inflammation in lung diseases. However, data on its inflammatory role in Cystic Fibrosis (CF) as part of the extracellular machinery driven by lung mesenchymal stem cells (MSCs)-derived extracellular vesicles (EVs) are missing. We obtained an in vitro model of CF-MSC by treating control human lung MSCs with a specific CFTR inhibitor. We characterized EVs populations derived from MSCs (ctr EVs) and CF-MSCs (CF-EVs) and analyzed their sphingolipid profile by LC-MS/MS. To evaluate their immunomodulatory function, we treated an in vitro human model of CF, with both EVs populations. Our data show that the two EVs populations differ for the average size, amount, and rate of uptake. CF-EVs display higher ceramide and dihydroceramide accumulation as compared to control EVs, suggesting the involvement of the de novo biosynthesis pathway in the parental CF-MSCs. Higher sphingomyelinase activity in CF-MSCs, driven by inflammation-induced ceramide accumulation, sustains the exocytosis of vesicles that export new formed pro-inflammatory ceramide. Our results suggest that CFTR dysfunction associates with an enhanced sphingolipid metabolism leading to the release of EVs that export the excess of pro-inflammatory Cer to the recipient cells, thus contributing to maintain the unresolved inflammatory status of CF

Connexin Expression in Human Minor Salivary Glands: An Immunohistochemical Microscopy Study

Connexins (Cxs) are transmembrane proteins involved in the formation of hemichannels and gap junctions (GJs). GJs are involved in various physiological functions, including secretion in glandular tissue. It has been demonstrated that Cx26, Cx32, and Cx43 are mainly expressed in glands, but no data are available in human salivary glands to date. The aim of our study was to investigate the presence and the localization of Cxs in human minor labial salivary glands. Immunofluorescence and immunoelectron microscopy were employed to evaluate the Cx26, Cx32, and Cx43 protein in human labial salivary gland biopsies (hLSGBs). RT-PCR was also used to detect their mRNA expression. Cx expression was found at both the mRNA and protein levels in all hLSGBs analysed. Cxs were observed at the level of the duct and acinar cells, as well as in myoepithelial cells. The localization of the three Cx types was very similar, suggesting colocalization of these Cxs in the same connexons. These results demonstrated the presence of Cxs in human salivary glands for the first time. Moreover, the few samples with primary Sjogren's Syndrome analysed only by immunofluorescence showed an alteration of the Cx expression, indicating that these proteins could be involved in salivary gland dysfunctions

Therapeutic Effect of Iron Citrate in Blocking Calcium Deposition in High Pi-Calcified VSMC: Role of Autophagy and Apoptosis

In chronic kidney disease (CKD), the first cause of mortality is cardiovascular disease induced mainly by vascular calcification (VC). Recently, iron-based phosphate binders have been proposed in advanced CKD to treat hyperphosphatemia. We studied the effect of iron citrate (iron) on the progression of calcification in high-phosphate (Pi) calcified VSMC. Iron arrested further calcification when added on days 7\u201315 in the presence of high Pi (1.30 \ub1 0.03 vs 0.61 \ub1 0.02; OD/mg protein; day 15; Pi vs Pi + Fe, p < 0.01). We next investigated apoptosis and autophagy. Adding iron to high-Pi-treated VSMC, on days 7\u201311, decreased apoptotic cell number (17.3 \ub1 2.6 vs 11.6 \ub1 1.6; Annexin V; % positive cells; day 11; Pi vs Pi + Fe; p < 0.05). The result was confirmed thorough analysis of apoptotic nuclei both in VSMCs and aortic rings treated on days 7\u201315 (3.8 \ub1 0.2 vs 2.3 \ub1 0.3 and 4.0 \ub1 0.3 vs 2.2 \ub1 0.2; apoptotic nuclei; arbitrary score; day 15; Pi vs Pi + Fe; VSMCs and aortic rings; p < 0.05). Studying the prosurvival axis GAS6/AXL, we found that iron treatment on days 9\u201314 counteracted protein high-Pi-stimulated down-regulation and induced its de novo synthesis. Moreover, iron added on days 9\u201315 potentiated autophagy, as detected by an increased number of autophagosomes with damaged mitochondria and an increase in autophagic flux. Highlighting the effect of iron on apoptosis, we demonstrated its action in blocking the H2O2-induced increase in calcification added both before high Pi treatment and when the calcification was already exacerbated. In conclusion, we demonstrate that iron arrests further high Pi-induced calcium deposition through an anti-apoptotic action and the induction of autophagy on established calcified VSMC

Suitability of nanoparticles to face benzo(A)pyrene-induced genetic and chromosomal damage in m. galloprovincialis. an in vitro approach

Benzo(a)pyrene (B(a)P) is a well-known genotoxic agent, the removal of which from environmental matrices is mandatory, necessitating the application of cleaning strategies that are harmless to human and environmental health. The potential application of nanoparticles (NPs) in the remediation of polluted environments is of increasing interest. Here, specifically designed NPs were selected as being non-genotoxic and able to interact with B(a)P, in order to address the genetic and chromosomal damage it produces. A newly formulated pure anatase nano-titanium (nano-TiO2), a commercial mixture of rutile and anatase, and carbon black-derived hydrophilic NPs (HNP) were applied. Once it had been ascertained that the NPs selected for the work did not induce genotoxicity, marine mussel gill biopsies were exposed in vitro to B(a)P (2 µg/mL), alone and in combination with the selected NPs (50 µg/mL nano-TiO2, 10 µg/mL HNP). DNA primary reversible damage was evaluated by means of the Comet assay. Chromosomal persistent damage was assessed on the basis of micronuclei frequency and nuclear abnormalities by means of the Micronucleus-Cytome assay. Transmission Electron Microscopy (TEM) was performed to investigate the mechanism of action exerted by NPs. Pure Anatase n-TiO2 was found to be the most suitable for our purpose, as it is cyto-and genotoxicity free and able to reduce the genetic and chromosomal damage associated with exposure to B(a)P