Circadian rhythm and melatonin in liver carcinogenesis: updates on current findings

: Liver cancer, including hepatocellular carcinoma and cholangiocarcinoma, can be devastating if not treated early. The risk factors of liver cancer include alcoholic liver disease, non-alcoholic fatty liver disease, disruption of melatonin levels, and dysregulated circadian rhythm. The circadian rhythm is a 24-hour biological clock that regulates the physiological activities at both central and peripheral levels. Its molecular mechanism exists in every cell in mammals. Disruption of the circadian rhythm has found in liver cancers as an independent risk factor. This review summarized the most recent findings about the molecular mechanisms of circadian rhythm, the crosstalk between core clock genes and melatonin, as well as the role of circadian rhythm and melatonin played in chronic liver diseases and liver cancer. Finally, we discussed the potential clinical application of circadian rhythm and melatonin for the treatment of liver cancer and discussed future perspectives of how understanding the circadian rhythm in liver cancer progression could provide new clinical applications for liver cancer treatment and diagnosis

Mast cells selectively target large cholangiocytes during biliary injury via H2HR-mediated cAMP/pERK1/2 signaling

Bile ducts are heterogenous in structure and function, and primary sclerosing cholangitis (PSC) damages specific bile ducts leading to ductular reaction (DR), mast cell (MC) infiltration, increased histamine release, inflammation, and fibrosis. Bile duct ligation (BDL) induces large duct damage via cyclic adenosine monophosphate (cAMP)/extracellular signal-related protein kinase (ERK) signaling, and large cholangiocytes express H2 histamine receptor (H2HR). We evaluated how MCs interact with large cholangiocytes during cholestasis. Male wild-type (WT) and MC-deficient (Kit(W-sh)) mice 10-12 weeks of age were subjected to BDL for 7 days. Select Kit(W-sh) mice were injected with MCs pretreated with control or H2HR antagonist (ranitidine, 25 mu m, 48 h) via tail vein injection. In vitro, MC migration toward small mouse cholangiocytes (SMCCs) and large mouse cholangiocytes (LMCCs) treated with lipopolysaccharide or histamine (+/- ranitidine) was measured. LMCCs were stimulated with MC supernatants pretreated with control, alpha-methyl-dl-histidine (to block histamine release), or ranitidine. Liver damage, large duct DR/senescence, inflammation, fibrosis, and cAMP/ERK immunoreactivity increased in BDL WT and Kit(W-sh)+MC mice but decreased in BDL Kit(W-sh) and Kit(W-sh)+MC-H2HR mice. In vitro, MCs migrate toward damaged LMCCs (but not SMCCs) blocked by inhibition of H2HR. Loss of MC histamine or MC-H2HR decreases LMCC proliferation, senescence, H2HR, and cAMP/ERK levels. Human PSC livers have increased MC number found near DR, senescent ducts, and H2HR-positive ducts. Conclusion: Infiltrating MCs preferentially interact with large ducts via H2HR signaling promoting biliary and liver damage. Mediation of MCs may be a therapeutic strategy for PSC

Amelioration of Ductular Reaction by Stem Cell Derived Extracellular Vesicles in MDR2 Knockout Mice via Lethal-7 microRNA

Cholangiopathies are diseases that affect cholangiocytes, the cells lining the biliary tract. Liver stem cells (LSCs) are able to differentiate into all cells of the liver and possibly influence the surrounding liver tissue by secretion of signaling molecules. One way in which cells can interact is through secretion of extracellular vesicles (EVs), which are small membrane-bound vesicles that contain proteins, microRNAs (miRNAs), and cytokines. We evaluated the contents of liver stem cell–derived EVs (LSCEVs), compared their miRNA contents to those of EVs isolated from hepatocytes, and evaluated the downstream targets of these miRNAs. We finally evaluated the crosstalk among LSCs, cholangiocytes, and human hepatic stellate cells (HSCs). We showed that LSCEVs were able to reduce ductular reaction and biliary fibrosis in multidrug resistance protein 2 (MDR2)−/− mice. Additionally, we showed that cholangiocyte growth was reduced and HSCs were deactivated in LSCEV-treated mice. Evaluation of LSCEV contents compared with EVs derived from hepatocytes showed a large increase in the miRNA, lethal-7 (let-7). Further evaluation of let-7 in MDR2−/− mice and human primary sclerosing cholangitis samples showed reduced levels of let-7 compared with controls. In liver tissues and isolated cholangiocytes, downstream targets of let-7 (identified by ingenuity pathway analysis), Lin28a (Lin28 homolog A), Lin28b (Lin28 homolog B), IL-13 (interleukin 13), NR1H4 (nuclear receptor subfamily 1 group H member 4) and NF-κB (nuclear factor kappa B), are elevated in MDR2−/− mice, but treatment with LSCEVs reduced levels of these mediators of ductular reaction and biliary fibrosis through the inhibition of NF-κB and IL-13 signaling pathways. Evaluation of crosstalk using cholangiocyte supernatants from LSCEV-treated cells on cultured HSCs showed that HSCs had reduced levels of fibrosis and increased senescence. Conclusion: Our studies indicate that LSCEVs could be a possible treatment for cholangiopathies or could be used for target validation for future therapies

The oral and gut microbiota: beyond a short communication

Introduction. The current treatment and prevention of oral disorders, dental caries, periodontal and gum diseases, follow a very non-specific control of plaque as the main causative factor. The main therapeutically approach is carried out on the sole perspective to keep the levels of oral bacteria in an acceptable range compatible with one-way vision of oral-mouth health, as something completely separated from a systemic microbial homeostasis (dysbiosis) concomitant present in the gut. A sealed compartmental view which sees separate and incommunicable responses to a specific condition without considering the presence of interacting confounding factors can negatively influence the diagnosis a diseases and of course its progression. A general non-specific antimicrobial with more general antiplaque therapy based mainly on oral care products together with surgery interventions represent at the moment the only mechanical responses in treating oral diseases. Material and method. The present paper is a narrative review concening interractions between oral and gut microbiota, with a focus on the interdisciplinary approach in antimicrobial treatment. Pubmed, Cochrane Library database were used for searching engines. Key words used were as follows: "inflammatory bowel syndrome (IBS)", "ulcerative colitis", "oral dysbiosis", "gut dysbiosis", "probiotics", "periodontitis". Results and discussions. Literature research showed that there are few issues to be discussed the ever increasing resistance to antibiotics, the high consumption of industrial food and sugars and their negatively effect on gut and oral microbiota. There is a need to highlight and develop a novel philosophical approach in the treatments for oral diseases that will necessarily involve non-conventional antimicrobial solutions. Such approaches should preferably reduce the consumption of both intestinal and oral microbiota, that are intimately connected and host approximately well over 1000 different species of bacteria at 108–109 bacteria per mL of mucous and saliva. Preventive approaches based upon the restoration of the microbial ecological balance, rather than elimination of the disease associated species, have been proposed. Conclusions. Having both oral-gut microbiota screened is an essential moment that influence the healthy immune modulatory and regenerative capacity of the body and, the new proposed formula integrates a wider screen on the patients where oral condition is strictly evaluated together with gut screen; therefore any proposed treatment will be inevitably sustained by the use of prebiotics and probiotics to promote health-associated bacterial growth. Keywords: inflammatory bowel syndrome (IBS), ulcerative colitis, oral dysbiosis, gut dysbiosis, probiotics, periodontitis

Mast cells in liver disease progression: An update on current studies and implications

: Mast cells (MCs) induce the progression of liver diseases including, but not limited to, hepatocellular carcinoma (HCC), cholangiocarcinoma (CCA), alcoholic and non-alcoholic fatty liver disease (ALD/NAFLD), primary biliary cholangitis (PBC), and primary sclerosing cholangitis (PSC). The effects of MCs during disease progression includes alterations in ductular reaction, steatosis, hepatic fibrosis and inflammation. In addition, there is significant crosstalk between MCs, MC mediators (histamine, tryptase, chymase) and MC-derived cytokines (transforming growth factor beta, tumor necrosis factor alpha, interleukins). Studies have been performed in rodent models, cultured cells, and human tissues to demonstrate the intracellular signaling implications of MC infiltration during liver disease. Targeting MCs may offer novel therapeutic strategies to treat liver disease. Our concise review will encompass the most recent studies involving MCs, their mediators and liver disease with the overall goal to inform the reader about the diverse role of these inflammatory immune cells in liver damage

Essential oils utility implications in symptomatic Burning Mouth Syndrome

Introduction. Burning mouth syndrome (BMS) is a clinical condition characterized by the presence of chronic pain in absence of clinically visible lesions of the oral mucosa. The etiology is uncertain and the therapeutic strategies still controversial. The objective of this prospective study is to analyze the efficacy of essential oils-based mouthwashes in the therapy of BMS. Material and method. This study included 16 patients affected by BMS who were treated with essential oils-based mouthwashes and glucose solution on alternated days for 30 days. Symptomatology was evaluated after 15, 30 and 90 days. Results and discussions. A the end of the treatment, most of the patients (67%) referred an improvement of symptoms up to complete remission in 90 days. Conclusions. Based on this study, essential oils-based mouthwashes could represent a valid aid in the treatment of BMS. Further studies are necessary in order to identify effective and standardized therapeutic protocols. Keywords: Burning Mouth Syndrome; oral rinse; essential oils; therapeutic strategies

Biostimulation with low-level laser therapy and its effects on soft and hard tissue regeneration. Literature review

Objective. Low-Level Laser Therapy encourages the healing process, reduces inflammation and pain. The aim of this study is to identify the impact of Low-Level Laser Therapy on tissue regeneration with special attention to hard tissues and to compare the effect of several wave lengths in the proliferation and differentiation of cells. Methods. The keywords used were “bone regeneration”, “laser therapy”, “photobiomodulation” OR “bio-stimulation”, “Low-Level Laser therapy” OR “LLLT”, “osteoblast proliferation” AND “differentiation”. Results. The bio-stimulation with Low-Level Laser Therapy also seems to interfere with the osseous integration of implants, by increasing its adherence on the bone-implant surfaces. Evidence has shown that Low-Level Laser Therapy influences the cellular proliferation and differentiation. Conclusions. Low-Level Laser Therapy is a promising therapy in the field of regeneration, but further studies are needed in order to define the standard protocol

Impacted Central Incisors in the Upper Jaw in an Adolescent Patient: Orthodontic-Surgical Treatment—A Case Report

The inclusion of both maxillary permanent central incisors is uncommon. This condition compromises face aesthetics, phonation andmasticatory function. Therefore, early diagnosis is essential to avoid complications and failures. There are various reasons for inclusion, but supernumerary teeth are the leading cause. Early causes of removal and rapid expansion of the palate determine a high probability of success with the spontaneous eruption of the impacted elements. However, it is often necessary to proceed with a surgical–orthodontic treatment. The inclination of teeth in relation to the midline and the root maturation degree determine prognosis and therapeutic timing. In this case report, the orthopantomogram (OPG) X-ray of a 9-year-old boy revealed two impacted supernumerary teeth in the anterior maxillary region, preventing the eruption of the permanent upper central incisors. The impacted supernumerary teeth were surgically removed at different times. A straight wire multibrackets technique associated with a fixed palatal appliance was used. The palatal appliance featured an osteomucous resin support at the level of the retroincisal papilla. Subsequently, surgical exposure was carried out using the closed eruption technique and elastictraction, bringing 11 and 21 back into the arch

Downregulation of p16 Decreases Biliary Damage and Liver Fibrosis in the Mdr2 / Mouse Model of Primary Sclerosing Cholangitis

Biliary senescence and hepatic fibrosis are hallmarks of cholangiopathies including primary sclerosing cholangitis (PSC). Senescent cholangiocytes display senescence-associated secretory phenotypes [SASPs, e.g., transforming growth factor-1 (TGF-1)] that further increase biliary senescence (by an autocrine loop) and trigger liver fibrosis by paracrine mechanisms. The aim of this study was to determine the effect of p16 inhibition and role of the TGF-1/microRNA (miR)-34a/sirtuin 1 (SIRT1) axis in biliary damage and liver fibrosis in the Mdr2/ mouse model of PSC. We treated (i) in vivo male wild-type (WT) and Mdr2/ mice with p16 Vivo-Morpholino or controls before measuring biliary mass [intrahepatic bile duct mass (IBDM)] and senescence, biliary SASP levels, and liver fibrosis, and (ii) in vitro intrahepatic murine cholangiocyte lines (IMCLs) with small interfering RNA against p16 before measuring the mRNA expression of proliferation, senescence, and fibrosis markers. p16 and miR-34a increased but SIRT1 decreased in Mdr2/ mice and PSC human liver samples compared to controls. p16 immunoreactivity and biliary senescence and SASP levels increased in Mdr2/ mice but decreased in Mdr2/ mice treated with p16 Vivo-Morpholino. The increase in IBDM and hepatic fibrosis (observed in Mdr2/ mice) returned to normal values in Mdr2/ mice treated with p16 Vivo-Morpholino. TGF-1 immunoreactivity and biliary SASPs levels were higher in Mdr2/ compared to those of WT mice but returned to normal values in Mdr2/ mice treated with p16 Vivo-Morpholino. The expression of fibrosis/senescence markers decreased in cholangiocytes from Mdr2/ mice treated with p16 Vivo-Morpholino (compared to Mdr2/ mice) and in IMCLs (after p16 silencing) compared to controls. Modulation of the TGF-1/miR-34a/SIRT1 axis may be important in the management of PSC phenotypes

Elastodontic Therapy of Hyperdivergent Class II Patients Using AMCOP® Devices: A Retrospective Study

Background: The management of a hyperdivergent growth pattern is one of the most challenging in orthodontics and different treatments are advocated. The present study analyses the effectiveness of elastodontic therapy with AMCOP® devices in treating children with hyperdivergent class II malocclusion and the effect on the upper airway patency. Methods: The study group included 21 patients (10 males and 11 females, mean age 8.22 ± 1.17 years) with a hyperdivergent growth and a class II malocclusion treated with AMCOP® devices. Cephalometric analysis was performed before treatment (T0) and after treatment (T1). Results: After treatment, the cephalometric analysis revealed a correction of the class II malocclusion and a modification of the growth pattern with a divergence reduction. The improvement of the upper airway space was also observed. Conclusion: The elastodontic therapy effectively corrected hyperdivergent class II malocclusion in growing patients over a short period