The lipidomic and inflammatory profiles of visceral and subcutaneous adipose tissues are distinctly regulated by the SGLT2 inhibitor empagliflozin in Zucker diabetic fatty rats.

The pharmacological inhibition of sodium-glucose cotransporter 2 (SGLT2) has emerged as a treatment for patients with type 2 diabetes mellitus (T2DM), cardiovascular disease and/or other metabolic disturbances, although some of the mechanisms implicated in their beneficial effects are unknown. The SGLT2 inhibitor (SGLT2i) empagliflozin has been suggested as a regulator of adiposity, energy metabolism, and systemic inflammation in adipose tissue. The aim of our study was to evaluate the impact of a 6-week-empagliflozin treatment on the lipidome of visceral (VAT) and subcutaneous adipose tissue (SAT) from diabetic obese Zucker Diabetic Fatty (ZDF) rats using an untargeted metabolomics approach. We found that empagliflozin increases the content of diglycerides and oxidized fatty acids (FA) in VAT, while in SAT, it decreases the levels of several lysophospholipids and increases 2 phosphatidylcholines. Empagliflozin also reduces the expression of the cytokines interleukin-1 beta (IL-1ß), IL-6, tumor necrosis factor-alpha (TNFα), monocyte-chemotactic protein-1 (MCP-1) and IL-10, and of Cd86 and Cd163 M1 and M2 macrophage markers in VAT, with no changes in SAT, except for a decrease in IL-1ß. Empagliflozin treatment also shows an effect on lipolysis increasing the expression of hormone-sensitive lipase (HSL) in SAT and VAT and of adipose triglyceride lipase (ATGL) in VAT, together with a decrease in the adipose content of the FA transporter cluster of differentiation 36 (CD36). In conclusion, our data highlighted differences in the VAT and SAT lipidomes, inflammatory profiles and lipolytic function, which suggest a distinct metabolism of these two white adipose tissue depots after the empagliflozin treatment.

In Vivo Effects of Conditioned Medium from Human Uterine Cervical Stem Cells in an Ovarian Cancer Xenograft Mouse Model.

BACKGROUND/AIM: Ovarian cancer is the most lethal of all gynecological cancers, despite advances in surgical techniques and medical treatments. During the last years, therapies based on mesenchymal stem cells and particularly their secretome (conditioned medium, CM) have emerged as promising treatments for various types of tumors. MATERIALS AND METHODS: In the present study, we evaluated the in vivo antitumor effect of human uterine cervical stem cell conditioned medium (hUCESC-CM) after intraperitoneal administration in an ovarian cancer mouse model. RESULTS: We found that intraperitoneal injection of hUCESC-CM in immunodeficient mice, injected fifty days previously with the human ovarian adenocarcinoma SKOV-3 cell line, significantly reduced abdominal tumor growth, and significantly increased overall survival, compared to control mice. CONCLUSION: hUCESC-CM could be an alternative approach to intraperitoneal treatment of ovarian cancer, either administered alone and/or with conventional chemotherapy.

Is there a correlation between HER2 gene amplification level and response to neoadjuvant treatment with trastuzumab and chemotherapy in HER2-positive breast cancer?

There are contradictory data regarding the correlation between HER2 amplification level determined by in situ hybridization and evolution after treatment with anti-HER2 therapies. The aim of this study was to correlate quantitative results of FISH (ratio HER2/CEP17 and number of HER2 signals/nucleus) with pathological response achieved after neoadjuvant treatment with trastuzumab and chemotherapy. For this purpose, we analysed 100 consecutive HER2-positive cases of breast carcinoma treated with neoadjuvant therapy. HER2 amplification determined by FISH was found in 92 of the 100 cases studied. pCR was obtained in 58% of the patients whose tumours presented amplification. In contrast, no pCR was obtained in the 8 patients with non-amplified tumours. A significant direct correlation between HER2 high amplification (HER2/CEP17 ratio > 5 or HER2 signals/nucleus > 10) and pCR was found. In conclusion, HER2 amplification levels are clinically relevant because they provide oncologists with valuable information on the possibilities of achieving pCR after neoadjuvant treatment.

Merkel cells of human oral mucosa express the pluripotent stem cell transcription factor Sox2.

Merkel cells are neuroendocrine cells associated to a neural sensitive ending and localized primarily in the epidermis, although they are also found in oral mucosa. Sox2 or SRY-box2 is a key transcription factor important in the maintenance of embryonic neural crest stem cell pluripotency. Sox2 has been described in Merkel cells of skin and in Merkel cell carcinomas, but not specifically in oral Merkel cells. The aims of the present study were to analyze the density of Merkel cells in human oral mucosa and to study the expression of Sox2 in these cells. For these purposes, immunohistochemical analyses for Sox2 and CK20 (the best marker for Merkel cells) were automatically performed on sections of normal human oral mucosa. Double immunofluorescence for Sox2 and CK20 was also performed. To analyze the density of Merkel cells, CK20 positive cells were counted in each sample and the length of the epithelial apical edge was measured (cells/mm). Merkel cells, demonstrated by CK20 immunoreactivity, were found in 95% of oral mucosa specimens studied (n=21). Mean density of Merkel cells in oral mucosa was 1.71±2.34 cells/mm. Sox2 immunoreactivity was found in the nuclei of scattered cells located at the basal layer. Serial sections immunostained for Sox2 and CK20 showed that Sox2-positive cells of oral mucosa coexpressed CK20, confirming that they were Merkel cells. Immunofluorescence for Sox2 and CK20 showed colocalization of both markers, demonstrating that virtually all oral Merkel cells expressed Sox2. This transcription factor could play a role in Merkel cell maturation and maintenance.

Association between hospital interval and survival in patients with oral cancer: A waiting time paradox.

BACKGROUND: In early diagnosis studies on symptomatic cancer, survival was the most recommended outcome. The magnitude and impact of the patient interval and primary care interval is well-known in oral cancer; however, the hospital interval and its influence on surviving this neoplasia are not well known. AIMS: To quantify the interval between the first contact with the specialist and the start of treatment for patients with oral cancer and to evaluate whether there was a link between this interval and disease survival. METHODS: We designed a hospital-based study that included 228 patients diagnosed with oral/oropharyngeal squamous cell carcinoma between 1998 and 2008 at A Coruña University Hospital (Spain) who were followed up until 2016. The data were extracted retrospectively from hospital medical charts. The study interval was defined in the context of the "pathways to treatment" model as the interval from the first specialist visit (start point) to the start of treatment (end point). We calculated the total interval (from first symptom to treatment) to evaluate the relative length of the hospital interval, and we considered the variables age, sex, location, comorbidity and tumour classification stage. Survival time was defined as the interval from the first treatment to death or censoring. RESULTS: The median hospital interval was 20 days, with an interquartile range of 15-29.1 days. The most relevant prognostic variable was the tumour stage (III-IV: Exp. ß = 2.8, p = 0.001). The hospital interval was part of the multivariate model, and its association with mortality showed a V-shaped association, where patients with short hospital intervals (3-18 days) and those with long hospital intervals (26-55 days) had significantly higher mortality than those with medium hospital intervals (19-25 days). CONCLUSION: The hospital interval represents a relevant interval for the patient's path towards treatment, has prognostic implications and is subject to a severity bias (waiting time paradox) that should be avoided.

MCH Regulates SIRT1/FoxO1 and Reduces POMC Neuronal Activity to Induce Hyperphagia, Adiposity, and Glucose Intolerance.

Melanin-concentrating hormone (MCH) is an important regulator of food intake, glucose metabolism, and adiposity. However, the mechanisms mediating these actions remain largely unknown. We used pharmacological and genetic approaches to show that the sirtuin 1 (SIRT1)/FoxO1 signaling pathway in the hypothalamic arcuate nucleus (ARC) mediates MCH-induced feeding, adiposity, and glucose intolerance. MCH reduces proopiomelanocortin (POMC) neuronal activity, and the SIRT1/FoxO1 pathway regulates the inhibitory effect of MCH on POMC expression. Remarkably, the metabolic actions of MCH are compromised in mice lacking SIRT1 specifically in POMC neurons. Of note, the actions of MCH are independent of agouti-related peptide (AgRP) neurons because inhibition of γ-aminobutyric acid receptor in the ARC did not prevent the orexigenic action of MCH, and the hypophagic effect of MCH silencing was maintained after chemogenetic stimulation of AgRP neurons. Central SIRT1 is required for MCH-induced weight gain through its actions on the sympathetic nervous system. The central MCH knockdown causes hypophagia and weight loss in diet-induced obese wild-type mice; however, these effects were abolished in mice overexpressing SIRT1 fed a high-fat diet. These data reveal the neuronal basis for the effects of MCH on food intake, body weight, and glucose metabolism and highlight the relevance of SIRT1/FoxO1 pathway in obesity.

POU1F1 transcription factor promotes breast cancer metastasis via recruitment and polarization of macrophages.

Cancer progression requires cells surrounding tumors be reeducated and activated to support tumor growth. Oncogenic signals from malignant cells directly influence stromal composition and activation, but the factors mediating this communication are still not well understood. We have previously shown that the transcription factor POU class 1 homeobox 1 (POU1F1), also known as Pit-1, induces profound changes on neoplastic cell-autonomous processes favoring metastasis in human breast cancer. Here we describe for the first time Pit-1-mediated paracrine actions on macrophages in the tumor microenvironment by using cell lines in vitro, zebrafish and mouse models in vivo, and samples from human breast cancer patients. Through the release of CXCL12, Pit-1 in tumor cells was found to mediate the recruitment and polarization of macrophages into tumor-associated macrophages (TAMs). In turn, TAMs collaborated with tumor cells to increase tumor growth, angiogenesis, extravasation and metastasis to lung. Our data reveal a new mechanism of cooperation between tumor cells and macrophages favoring metastasis and poor clinical outcome in human breast cancer, which suggests that Pit-1 and CXCL12 should be further studied as potential prognostic and therapeutic indicators. © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Mechanical characteristics of the maxillary sinus Schneiderian membrane ex vivo.

OBJECTIVES: It has been speculated that certain Schneiderian membrane thickness (SMT) might be more prone to perforation. This investigation was aimed at studying the mechanical characteristics of the Schneiderian membrane under one- and two-dimensional tests and their correlation to the histological SMT in human samples. MATERIAL AND METHODS: Sixteen Schneiderian membranes were collected from 11 cadaver heads treated with Thiel's embalming method. The samples were processed and analyzed clinically and histologically. One-dimensional maximum elongation until perforation and two-dimensional resistance to ball penetration were performed after the biopsy. Data was analyzed by using the Wilcoxon rank test and the Spearman's rank correlation. RESULTS: The histological SMT was 1.36 ± 0.42 mm, whereas the clinical thickness was 0.27 ± 0.21 mm, yielding statistical significance (p = 0.000). The resistance under ball penetration was 0.59 ± 0.43 N and the mean maximum elongation in the one-dimension test 11.19 ± 7.14 mm. Expressed in percentage, the mean stretch was 241.36 ± 227.97% (range 31.5 up to 947%). A weak positive correlation was found between the ball penetration test and the SMT (r = 0.10, p = 0.711), while a weak negative correlation was found between stretching test and the SMT (r = -0.021, p = 0.94). CONCLUSIONS: Mechanical tests seem to indicate that SMT might not significantly predispose to Schneiderian membrane perforation. Hence, other anatomical and operator's factors should be considered of surpassing importance. CLINICAL RELEVANCE: Thinner SM might be more prone to perforation when detaching it from the maxillary sinus antrum; however, a thick membrane is not prevented to tear, as their resistance under elastic forces is not higher than thinner ones.

Pseudoepitheliomatous hyperplasia after diode laser oral surgery. An experimental study.

BACKGROUND: to examine the process of epithelial reparation in a surgical wound caused by diode laser. MATERIAL AND METHODS: An experimental study with 27 Sprage-Dawley rats was undertaken. The animals were randomly allocated to two experimental groups, whose individuals underwent glossectomy by means of a diode laser at different wattages, and a control group treated using a number 15 scalpel blade. The animals were slaughtered at the 2nd, 7th, and 14th day after glossectomy. The specimens were independently studied by two pathologists (blinded for the specimens' group). RESULTS: at the 7th day, re-epithelisation was slightly faster for the control group (conventional scalpel) (p=0.011). At the 14th day, complete re-epithelization was observed for all groups. The experimental groups displayed a pseudoepitheliomatous hyperplasia. CONCLUSIONS: it is concluded that, considering the limitations of this kind of experimental studies, early re-epithelisation occurs slightly faster when a conventional scalpel is used for incision, although re-epithelisation is completed in two weeks no matter the instrument used. In addition, pseudoepitheliomatous hyperplasia is a potential event after oral mucosa surgery with diode laser. Knowledge about this phenomenon (not previously described) may prevent diagnostic mistakes and inadequate treatment approaches, particularly when dealing with potentially malignant oral lesions.

Unifocal orofacial granulomatosis in retromolar mucosa: surgical treatment with Er,Cr:YSGG laser.

Orofacial granulomatosis is defined by permanent or recurrent swelling of orofacial tissues with different multiform and multifocal clinical patterns. An 11-year old boy presented with a 2-month history of mucosa enlargement. Intraoral examination revealed an erythematous, polylobulated, exophytic lesion with a smooth surface located in retromolar mucosa, non-tender and non-infiltratated to palpation. The diagnosis was inflammatory lesion compatible with pyogenic granuloma and laser excision was decided. Haematological parameters were within normal range, as well as chest Xrays. These findings lead to a diagnosis of non-symptomatic orofacial granulomatosis, whose early diagnosis can minimize the impact of systemic-related disorders, like Chron's disease. Key words:Laser, orofacial granulomatosis, childhood, oral lesions, diagnosis.

Membrane perforation in sinus floor elevation - piezoelectric device versus conventional rotary instruments for osteotomy: an experimental study.

PURPOSE: Sinus membrane perforation is the most common intraoperative complication of maxillary sinus floor elevation (MSFE) procedures and frequently causes postoperative problems. Piezoelectric devices have been claimed to reduce the frequency of membrane perforations although no clear evidence supports this view. MATERIALS AND METHODS: Ten surgeons with different expertise levels performed 80 MSFEs in selected lamb heads, with rotary and piezoelectric instruments following standard protocols. After the procedures, specimens were coded and perforations or tears determined through a microscope. RESULTS: No significant differences in terms of thickness either of the sinus lateral wall (xi -xj = 73.2; 95% confidence interval [CI] = 45.3-191.8) or the membrane (xi -xj = 24.2; 95% CI = -29.4 to 77.9) were identified between the specimens allocated to each group. Nine membrane perforations (11.2%) occurred during the study, all within the lower expertise group. Membrane elevation by hand instruments caused five perforations (40%) in the rotary instrument group and one in the piezoelectric group. Expert surgeons produced no membrane perforations, the size of the antrostomy that was smaller in the piezoelectric group being the only significant difference between the rotary and piezoelectric groups. CONCLUSIONS: The use of piezoelectric material for MSFE reduces the frequency of membrane perforation among surgeons with a limited experience.

Lamb ex vivo model for training in maxillary sinus floor elevation surgery: a comparative study with human standards.

BACKGROUND: Many models have been suggested for surgical training in sinus floor elevation (SFE), but information on key features that provide feedback to the trainee is scarce. The aim of this study is to compare the thickness of the lateral wall of the maxillary sinus (TLWMS) and the thickness of the Schneiderian membrane (TSM) between an animal model and the human standard. METHODS: The observational study used 20 fresh lamb heads. Cone-beam computed tomography (CT) was performed for recording the following: TLWMS, sinus lateral floor width, distance from the lateral border of the anteroposterior bone crest to the sinus wall, and height of lateral maxillary sinus. Thirty human radiologic exploration cone-beam CTs were used for comparisons. The sinus membrane thickness was measured at three points per section and compared to those of 10 formalin-preserved human membranes. RESULTS: TLWMS in the animal model scored 2.25 ± 0.3 mm and was thicker than that of humans with subsinusal edentulism (Xi - Xj = 0.3; 95% confidence interval = 0.1 to 0.4), although this difference was not considered clinically relevant. The average thickness of the sinus membrane was 485.5 ± 137.1 µm with no significant differences between sides (P = 0.12) or between the thickness of human and lamb Schneiderian membranes. CONCLUSIONS: The model based on lambs is potentially useful for training in SFE techniques because of the similarities in the TLWMS and particularly the TSM. Additional studies are needed to validate this model within a teaching environment.

Unsuspected foreign body aspiration.

Aspiration of foreign bodies is a serious complication that may occur during the course of dental treatment. A case report of a 60-year-old man with recurrent pneumonia is presented. One year after the onset of his initial symptoms, a hard substance that made a complete mold of the bronchial tree at the inferior right lobe was extracted with a rigid bronchoscopy. This green material of elastic consistency was dental impression material (polyvinylsiloxane). On careful questioning, the patient indicated that he had dental impressions taken 2 months before the onset of the symptoms. Surgery was indicated and lobectomy of the inferior right lobe was performed without incident. To avoid this complication, some preventive precautions such as identifying high-risk patients; using rubber dam; tethering any small instrument with a ligature; placing a gauze screen to protect the oropharynx in sedated patients; and using custom impression trays to minimize the amount of impression material required have been suggested. In case of a suspected aspiration, the patient must be referred to appropriate medical care.