Myeloid AMPK signaling restricts fibrosis but is not required for metformin improvements during CDAHFD-induced NASH in mice.

Metabolic programming underpins inflammatory processes of immune cells. In the context of chronic liver disease, liver macrophage activation and response to hepatocellular damage is dependent on profound metabolic changes. Here, we sought to identify the role of an important metabolic regulator, AMP-activated protein kinase (AMPK), specifically within myeloid cells during the progression of non-alcoholic steatohepatitis (NASH) and whether treatment with metformin, a first line therapy for diabetes and activator of AMPK could stem disease progression. Male and female Prkaa1fl/fl/Prkaa2fl/fl (Flox) control and Flox-LysM-Cre+ (MacKO) mice were fed a low-fat control or a choline-deficient, amino acid defined 45% Kcal high fat diet (CDAHFD) for 8 weeks, where metformin was introduced in the drinking water (50 or 250 mg/kg/day) for the last 4 weeks. Hepatic steatosis and fibrosis were dramatically increased in response to CDAHFD-feeding compared to low-fat control. While myeloid AMPK signaling had no effect on markers of hepatic steatosis or circulating markers, fibrosis as measured by total liver collagen was significantly elevated in livers from MacKO mice, independent of sex. Although treatment with 50 mg/kg/day metformin had no effect on any parameter, intervention with 250 mg/kg/day metformin completely ameliorated hepatic steatosis and fibrosis in both male and female mice. While the protective effect of metformin was associated with lower final body weight, decrease expression of lipogenic and Col1a1 transcripts, it was independent of myeloid AMPK signaling. These results suggest that endogenous AMPK signaling in myeloid cells, both liver-resident and infiltrating, acts to restrict fibrogenesis during CDAHFD-induced NASH progression but is not the mechanism by which metformin improves markers of NASH.

Selective outcome reporting in randomized clinical trials using the third molar surgery model.

Selective outcome reporting (SOR) can threaten the validity of results found in clinical trials. Some studies in the literature have analyzed SOR in dentistry, but there is no study that has observed SOR in clinical trials in oral and maxillofacial surgery. Impacted third molar surgery is one of the most used models in clinical trials to study mainly analgesic and anti-inflammatory drug interventions. Our study aimed to evaluate the prevalence of SOR in publications employing the third molar extraction clinical trial model, and to verify whether there was an association between the statistical significance of outcomes and other characteristics that could lead to SOR. A systematic search was performed on the ClinicialTrials.gov platform for randomized clinical trial protocols, using the condition of third molar extraction. The corresponding published articles were sourced in PubMed, Scopus, and Embase databases, and compared with the registered protocols regarding the methodological data, in terms of: sample calculation, primary outcome identification, end-point periods, insertion of new outcomes in the publication, and results of outcomes. 358 protocol records were retrieved; 87 presented their corresponding articles. SOR was identified in 28.74% of the publications, and had a significant relationship with changes in the protocol, insertions of new outcomes, and discrepancies in the types of study. General risk of bias was found to be low. There were associations between SOR and the discrepancies in terms of the type of study, the choice of new outcome, and changes in the history of protocol records. The prevalence of SOR in clinical research using the third molar extraction surgery model is moderate. The quality of the scientific reporting of the results and, consequently, the certainty of evidence relating to the intervention tested can be overstated, increasing the chances of misinterpretation by health professionals.

Schiff bases and their metal complexes to target and overcome (multidrug) resistance in cancer.

Overcoming multidrug resistance (MDR) is one of the major challenges in cancer therapy. In this respect, Schiff base-related compounds (bearing a R1R2CNR3 bond) gained high interest during the past decades. Schiff bases are considered privileged ligands for various reasons, including the easiness of their preparation and the possibility to form complexes with almost all transition metal ions. Schiff bases and their metal complexes exhibit many types of biological activities and are used for the treatment and diagnosis of various diseases. Until now, 13 Schiff bases have been investigated in clinical trials for cancer treatment and hypoxia imaging. This review represents the first collection of Schiff bases and their complexes which demonstrated MDR-reversal activity. The areas of drug resistance covered in this article involve: 1) Modulation of ABC transporter function, 2) Targeting lysosomal ABCB1 overexpression, 3) Circumvention of ABC transporter-mediated drug efflux by alternative routes of drug uptake, 4) Selective activity against MDR cancer models (collateral sensitivity), 5) Targeting GSH-detoxifying systems, 6) Overcoming apoptosis resistance by inducing necrosis and paraptosis, 7) Reactivation of mutated p53, 8) Restoration of sensitivity to DNA-damaging anticancer therapy, and 9) Overcoming drug resistance through modulation of the immune system. Through this approach, we would like to draw attention to Schiff bases and their metal complexes representing highly interesting anticancer drug candidates with the ability to overcome MDR.

ERK1/2 modulation in the central nervous system of male and female thiamine-deficient mice with amprolium.

Thiamine deficiency experimental models focus on using the pyrithiamine analog in male rodents, making the thiamine deficiency effects in females and the use of other thiamine antagonists, such as amprolium, unknown. We investigated the impact of thiamine deficiency with amprolium in the cerebral cortex and thalamus of male and female mice by evaluating the modulation of ERK1/2 phosphorylation. The animals were exposed for 20 days to thiamine-deficient chow with different doses of amprolium (20, 40, 60 and 80 mg/kg) and at different treatment periods (five, 10, 15 or 20 days) at a dose of 60 mg/kg. After treatments, ERK1/2 phosphorylation was analyzed by western blot. In male mice, we observed a progressive increase in ERK1/2 phosphorylation in both the cerebral cortex and thalamus in response to the dose of amprolium. In females, ERK1/2 phosphorylation did not progressively increase in response to the amprolium dosage. However, an increase in phosphorylation at the higher doses of 60 and 80 mg/kg was observed. We observed a more intense increase in ERK1/2 phosphorylation in males' cerebral cortex and thalamus from 10 days onwards. In females, the ERK1/2 modulation profiles were similar. The results show that thiamine deficiency induction with amprolium is efficient, compatible with other recognized models that use pyrithiamine, showing changes in cell signaling in the nervous system. The study showed differences in response to thiamine deficiency with amprolium between male and female mice in relation to ERK1/2 phosphorylation and demonstrated that females respond positively to the method and can also be used as model animals.

Comparison of different DNA preservation solutions for oral cytological samples.

OBJECTIVE: The objective of this study was to compare the DNA preservation capacity of buccal mucosa exfoliated cells when stored in different solutions under varying time and temperature conditions. DESIGN: DNA preservation solutions, including Dimethyl sulphoxide disodium-EDTA-saturated NaCl (DESS), Tris-EDTA-NaCl-Tween20 buffer (TENT), Nucleic Acid Preservation Buffer (NAP), and phosphate-buffered saline (PBS), were prepared. Buccal mucosa cells from a single patient were collected, dispensed into these solutions, and stored at room temperature (RT) and 4 °C for 24 h, 72 h, 30 days, 90 days, and 180 days. DNA was extracted using the salting-out method and the QIAamp DNA Mini Kit. DNA concentration and purity were determined using the QuBit device and NanoDrop, while DNA integrity was assessed using the Agilent 4200 TapeStation system. The ability to amplify the IFNA primer was also evaluated by PCR. RESULTS: The salting-out method yielded better concentration and purity results, with PBS, TENT, and DESS buffers demonstrating superior concentration values when stored at 4 °C, resulting in mean values exceeding 10 ng/µL for up to 30 days. DESS consistently exhibited the best integrity values over time for both temperature conditions. Amplification capacity was enhanced when samples were stored at 4 °C. When stored at RT, PBS achieved 100% amplification within 24 h. NAP yielded the poorest results. CONCLUSION: In the context of long-term preservation, the DESS buffer emerges as the most effective solution, maintaining requisite DNA quality and quantity standards for up to 30 days at RT and up to 3 months at 4 °C.

Choline metabolism underpins macrophage IL-4 polarization and RELMα up-regulation in helminth infection.

Type 2 cytokines like IL-4 are hallmarks of helminth infection and activate macrophages to limit immunopathology and mediate helminth clearance. In addition to cytokines, nutrients and metabolites critically influence macrophage polarization. Choline is an essential nutrient known to support normal macrophage responses to lipopolysaccharide; however, its function in macrophages polarized by type 2 cytokines is unknown. Using murine IL-4-polarized macrophages, targeted lipidomics revealed significantly elevated levels of phosphatidylcholine, with select changes to other choline-containing lipid species. These changes were supported by the coordinated up-regulation of choline transport compared to naïve macrophages. Pharmacological inhibition of choline metabolism significantly suppressed several mitochondrial transcripts and dramatically inhibited select IL-4-responsive transcripts, most notably, Retnla. We further confirmed that blocking choline metabolism diminished IL-4-induced RELMα (encoded by Retnla) protein content and secretion and caused a dramatic reprogramming toward glycolytic metabolism. To better understand the physiological implications of these observations, naïve or mice infected with the intestinal helminth Heligmosomoides polygyrus were treated with the choline kinase α inhibitor, RSM-932A, to limit choline metabolism in vivo. Pharmacological inhibition of choline metabolism lowered RELMα expression across cell-types and tissues and led to the disappearance of peritoneal macrophages and B-1 lymphocytes and an influx of infiltrating monocytes. The impaired macrophage activation was associated with some loss in optimal immunity to H. polygyrus, with increased egg burden. Together, these data demonstrate that choline metabolism is required for macrophage RELMα induction, metabolic programming, and peritoneal immune homeostasis, which could have important implications in the context of other models of infection or cancer immunity.

Synthesis of films based on chitosan and protic ionic liquids to be used as wound dressing on the oral mucosa.

Oral mucosal ulcerations expose connective tissue to different pathogens and this can progress to systemic infection. This study aimed to synthesize environmentally-friendly films with chitosan and protic ionic liquids, possessing mucoadhesive properties, activity against opportunistic microorganisms, enhanced malleability and mechanical resistance to be used as a wound dressing on the oral mucosa. Therefore, films with chitosan and 10, 35, and 50 % (wt/wt) of 2-hydroxy diethylammonium lactate, salicylate, and maleate protic ionic liquids were synthesized. Thickness measurements and mechanical properties analysis were performed. In addition, oral mucoadhesion, antimicrobial activity, and cytotoxicity properties were investigated. Results showed that the addition of 35wt% and 50wt% of all kinds of protic ionic liquids tested presented significant improvements in film thickness and mechanical properties. Films based on chitosan and the protic ionic liquid 2-hydroxy diethylammonium salicylate at percentages of 35 and 50wt% exhibited superior mucoadhesive properties, antimicrobial activity on opportunistic microorganisms and an improvement in their flexibility after immersion in synthetic saliva. Cytotoxicity results suggest that all kinds of chitosan/protic ionic liquids films tested are safe for intra-oral use. Therefore, the results of this study indicate that these materials could be good candidates for efficient and environmentally-friendly wound dressing films on the oral mucosa.

Thermoneutral housing does not accelerate metabolic dysfunction-associated fatty liver disease in male or female C57Bl/6J mice fed a Western diet.

Metabolic dysfunction-associated fatty liver disease (MAFLD) represents a growing cause of mortality and morbidity and encompasses a spectrum of liver pathologies. Although dozens of preclinical models have been developed to recapitulate stages of MAFLD, few achieve fibrosis using an experimental design that mimics human pathogenesis. We sought to clarify whether the combination of thermoneutral (TN) housing and consumption of a classical Western diet (WD) would accelerate the onset and progression of MAFLD. Male and female C57Bl/6J mice were fed a nutrient-matched low-fat control or Western diet (WD) for 16 wk. Mice were housed with littermates at either standard temperature (TS; 22°C) or thermoneutral-like conditions (TN; ∼29°C). Male, but not female, mice housed at TN and fed a WD were significantly heavier than TS-housed control animals. WD-fed mice housed under TN conditions had lower levels of circulating glucose compared with TS mice; however, there were select but minimal differences in other circulating markers. Although WD-fed TN males had higher liver enzyme and higher liver triglyceride levels, no differences in markers of liver injury or hepatic lipid accumulation were observed in females. Housing temperature had little effect on histopathological scoring of MAFLD progression in males; however, although female mice retained a level of protection, WD-TN conditions trended toward a worsened hepatic phenotype, which was associated with higher macrophage transcript expression and content. Our results indicate that interventions coupling TN housing and WD-induced MAFLD should be longer than 16 wk to accelerate hepatic steatosis and increase inflammation in both sexes of mice.NEW & NOTEWORTHY Mouse models leading to accelerated fatty liver onset are a useful translational tool. Here we show that coupling thermoneutral-like housing and Western diet feeding in mice for 16 wk does not lead to significant disease progression in either sex, though the molecular phenotype indicates priming of immune-related and fibrotic pathways.

Usability of Telehealth Systems for Noncommunicable Diseases in Primary Care From the COVID-19 Pandemic Onward: Systematic Review.

BACKGROUND: During the COVID-19 pandemic, telehealth was expanded without the opportunity to extensively evaluate the adopted technology's usability. OBJECTIVE: We aimed to synthesize evidence on health professionals' perceptions regarding the usability of telehealth systems in the primary care of individuals with noncommunicable diseases (NCDs; hypertension and diabetes) from the COVID-19 pandemic onward. METHODS: A systematic review was performed of clinical trials, prospective cohort studies, retrospective observational studies, and studies that used qualitative data collection and analysis methods published in English, Spanish, and Portuguese from March 2020 onward. The databases queried were MEDLINE, Embase, BIREME, IEEE Xplore, BVS, Google Scholar, and grey literature. Studies involving health professionals who used telehealth systems in primary care and managed patients with NCDs from the COVID-19 pandemic onward were considered eligible. Titles, abstracts, and full texts were reviewed. Data were extracted to provide a narrative qualitative evidence synthesis of the included articles. The risk of bias and methodological quality of the included studies were analyzed. The primary outcome was the usability of telehealth systems, while the secondary outcomes were satisfaction and the contexts in which the telehealth system was used. RESULTS: We included 11 of 417 retrieved studies, which had data from 248 health care professionals. These health care professionals were mostly doctors and nurses with prior experience in telehealth in high- and middle-income countries. Overall, 9 studies (82%) were qualitative studies and 2 (18%) were quasiexperimental or multisite trial studies. Moreover, 7 studies (64%) addressed diabetes, 1 (9%) addressed diabetes and hypertension, and 3 (27%) addressed chronic diseases. Most studies used a survey to assess usability. With a moderate confidence level, we concluded that health professionals considered the usability of telehealth systems to be good and felt comfortable and satisfied. Patients felt satisfied using telehealth. The most important predictor for using digital health technologies was ease of use. The main barriers were technological challenges, connectivity issues, low computer literacy, inability to perform complete physical examination, and lack of training. Although the usability of telehealth systems was considered good, there is a need for research that investigates factors that may influence the perceptions of telehealth usability, such as differences between private and public services; differences in the level of experience of professionals, including professional experience and experience with digital tools; and differences in gender, age groups, occupations, and settings. CONCLUSIONS: The COVID-19 pandemic has generated incredible demand for virtual care. Professionals' favorable perceptions of the usability of telehealth indicate that it can facilitate access to quality care. Although there are still challenges to telehealth, more than infrastructure challenges, the most reported challenges were related to empowering people for digital health. TRIAL REGISTRATION: PROSPERO International Prospective Register of Systematic Reviews CRD42021296887; https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=296887. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): RR2-10.21801/ppcrj.2022.82.6.

Hepatocyte-derived DPP4 regulates portal GLP-1 bioactivity, modulates glucose production, and when absent influences NAFLD progression.

Elevated circulating dipeptidyl peptidase-4 (DPP4) is a biomarker for liver disease, but its involvement in gluconeogenesis and metabolic associated fatty liver disease progression remains unclear. Here, we identified that DPP4 in hepatocytes but not TEK receptor tyrosine kinase-positive endothelial cells regulates the local bioactivity of incretin hormones and gluconeogenesis. However, the complete absence of DPP4 (Dpp4-/-) in aged mice with metabolic syndrome accelerates liver fibrosis without altering dyslipidemia and steatosis. Analysis of transcripts from the livers of Dpp4-/- mice displayed enrichment for inflammasome, p53, and senescence programs compared with littermate controls. High-fat, high-cholesterol feeding decreased Dpp4 expression in F4/80+ cells, with only minor changes in immune signaling. Moreover, in a lean mouse model of severe nonalcoholic fatty liver disease, phosphatidylethanolamine N-methyltransferase mice, we observed a 4-fold increase in circulating DPP4, in contrast with previous findings connecting DPP4 release and obesity. Last, we evaluated DPP4 levels in patients with hepatitis C infection with dysglycemia (Homeostatic Model Assessment of Insulin Resistance > 2) who underwent direct antiviral treatment (with/without ribavirin). DPP4 protein levels decreased with viral clearance; DPP4 activity levels were reduced at long-term follow-up in ribavirin-treated patients; but metabolic factors did not improve. These data suggest elevations in DPP4 during hepatitis C infection are not primarily regulated by metabolic disturbances.

Thiosemicarbazone Derivatives Developed to Overcome COTI-2 Resistance.

COTI-2 is currently being evaluated in a phase I clinical trial for the treatment of gynecological and other solid cancers. As a thiosemicarbazone, this compound contains an N,N,S-chelating moiety and is, therefore, expected to bind endogenous metal ions. However, besides zinc, the metal interaction properties of COTI-2 have not been investigated in detail so far. This is unexpected, as we have recently shown that COTI-2 forms stable ternary complexes with copper and glutathione, which renders this drug a substrate for the resistance efflux transporter ABCC1. Herein, the complex formation of COTI-2, two novel terminal N-disubstituted derivatives (COTI-NMe2 and COTI-NMeCy), and the non-substituted analogue (COTI-NH2) with iron, copper, and zinc ions was characterized in detail. Furthermore, their activities against drug-resistant cancer cells was investigated in comparison to COTI-2 and Triapine. These data revealed that, besides zinc, also iron and copper ions need to be considered to play a role in the mode of action and resistance development of these thiosemicarbazones. Moreover, we identified COTI-NMe2 as an interesting new drug candidate with improved anticancer activity and resistance profile.

Comparative Evaluation of MTA Sealers Biomineralization Ability: an in Vivo Study / Avaliação Comparativa da Capacidade de Biomineralização de Cimentos MTA: Estudo in Vivo

This research aimed to evaluate the biomineralization induced by different concentrations of MTA Flow® and compare it to MTA Angelus®. Fifteen male Wistar rats received subcutaneous implants containing the materials to be tested (MTA Flow® at putty, thick, and thin consistencies and MTAAngelus®) and empty tubes (control). After 7, 40 and 90 days, the animals were euthanized, and the implants were removed with the surrounding tissue. The presence of biomineralization was analyzed in light microscope by Von Kossa technique. The statistica l differences were considered for p < 0.05. Calcification areas were present in all the MTA Flow® and MTA Angelus® groups. In the control group, no mineralized areas were observed. MTA Angelus® and thin-MTA Flow® showed significant reduction in calcification as time went by. A significant increase in areas with calcification, proportional to the exposure time, was observed in putty-MTA Flow® and thick-MTA Flow®. MTA Angelus® and thin-MTA Flow® showed significantly higher calcification than thick-MTA Flow® in the shortest exposure time. Analysis of putty-MTA Flow® showed significantly higher calcification areas than MTA Angelus® and thin-MTA Flow® in the longest exposure time. MTA Flow® stimulated mineralization, which has varied according to the concentration. Besides, in longer periods, MTA Flow® biomineralization performance was higher than MTAAngelus®, especially in highest concentration. (AU)

Esse estudo teve como objetivo avaliar a biomineralização induzida por diferentes concentrações do MTA Flow® e compará-la ao MTA Angelus®. Quinze ratos Wistar machos receberam implantes subcutâneos contendo os materiais a serem testados (MTA Angelus® e MTA Flow® nas consistências pastosa, espessa e fluida) e tubos vazios (controle). Após 7, 40 e 90 dias, os animais foram eutanasiados e os implantes foram removidos juntamente com o tecido circundante. A presença de biomineralização foi analisada em microscópio de luz pela técnica de Von Kossa. Diferenças estatísticas foram consideradas para valores de p < 0,05. Áreas de calcificação estavam presentes em todos os grupos do MTA Flow® e MTA Angelus®. No grupo controle não foram observadas áreas mineralizadas. O MTA Angelus® e o MTA Flow® fluido apresentaram redução significativa na quantidade de calcificação ao longo do tempo. Um aumento significativo na quantidade de áreas calcificadas, proporcional ao tempo de exposição, foi observado no MTA Flow® pastoso e no MTA Flow® espesso. O MTA Angelus® e o MTA Flow® fluido apresentaram calcificação significativamente maior do que o MTA Flow® espesso no menor período de exposição. Análises contento o MTA Flow® pastoso demonstraram áreas de calcificação significativamente maior do que o MTA Angelus® e MTA Flow® fluido no maior tempo de exposição. O MTA Flow® induziu a formação de áreas mineralizadas, que variou de acordo com a concentração do cimento. Em períodos mais longos, o MTA Flow® apresentou desempenho superior ao MTA Angelus®, principalmente quando utilizado na maior concentração. (AU)

Investigating the antiproliferative activities of new CuII complexes with pyridine hydrazone derivatives of nalidixic acid.

To further explore the structural features and potential antibacterial and antitumor activities of polynuclear CuII coordination compounds with nalidixic acid (nx) derivatives, new complexes bearing nx hydrazones with N-pyridinyl moieties substituted at positions 2 and 3 (h2py and h3py) were synthesized. Complexes [Cu3(C18H16N5O2)2(C18H17N5O2)2(H2O)]4BF4∙H2O (1), and [Cu3(C18H16N5O2)2(C18H17N5O2)2(H2O)3]4BF4∙3H2O (%) (2) were synthesized using h2py and h3py with Cu(BF4)2∙nH2O as precursor, whereas the [Cu(C18H17N5O2)Cl2]∙0.5H2O complex (3) was synthesized with h2py and CuCl2∙2H2O. Crystallographic studies of complex 1, showed that coordination of hydrazones to CuII occurs by tridentate modes of type κ3(O,N,N') as well as bidentate modes of type κ2(O',N″). Complexes 1, 2 and 3 had their antiproliferative activities evaluated in vitro against a panel of tumor cells by the determination of GI50 values. Complexes 1 and 2 were more active than complex 3, suggesting an effect of the complex charge on their activities. The interactions of such complexes towards bovine serum albumin (BSA) and DNA plasmid (pGEX-4 T1) were investigated using fluorescence spectroscopy and gel electrophoresis. All complexes were shown to interact with the DNA model as metallonucleases, but no interaction with BSA was observed. DNA molecular docking of complex 1 encompassing both its trinuclear (TN) form and a possible mononuclear (MN) derivative suggests that naphthyridyl ring performs π-stacking interactions with DNA. The TN species were also shown to be possible minor groove binders.

TLR4 Expression in Ex-Lichenoid Lesions-Oral Squamous Cell Carcinomas and Its Surrounding Epithelium: The Role of Tumor Inflammatory Microenvironment.

Toll-like receptors (TLRs) regulate innate and adaptive immune responses. Moreover, TLRs can induce a pro-survival and pro-proliferation response in tumor cells. This study aims to investigate the expression of TLR4 in the epithelium surrounding oral squamous cell carcinomas (OSCC) in relation to its inflammatory microenvironment. This study included 150 human samples: 30 normal oral control (NOC), 38 non-lichenoid epithelium surrounding OSCC (NLE-OSCC), 28 lichenoid epithelium surrounding OSCC (LE-OSCC), 30 OSCC ex-non oral lichenoid lesion (OSCC Ex-NOLL), and 24 OSCC ex-oral lichenoid lesion (OSCC Ex-OLL). TLR4 expression was investigated by immunohistochemistry and the percentage of positive cells was quantified. In addition, a semiquantitative analysis of staining intensity was performed. Immunohistochemical analysis revealed that TLR4 is strongly upregulated in LE-OSCC as compared to normal control epithelium and NLE-OSCC. TLR4 expression was associated with the inflammatory environment, since the percentage of positive cells increases from NOC and NLE-OSCC to LE-OSCC, reaching the highest value in OSCC Ex-OLL. TLR4 was detected in the basal third of the epithelium in NLE-OSCC, while in LE-OSCC, TLR4 expression reached the intermediate layer. These results demonstrated that an inflammatory microenvironment can upregulate TLR4, which may boost tumor development.

The role of tumor acidification in aggressiveness, cell dissemination and treatment resistance of oral squamous cell carcinoma.

AIMS: To investigate the role of tumor acidification in cell behavior, migration, and treatment resistance of oral squamous cell carcinoma (OSCC). MAIN METHODS: The SCC4 and SCC25 cell lines were exposed to acidified (pH 6.8) cell culture medium for 7 days. Alternatively, a long-term acidosis was induced for 21 days. In addition, to mimic dynamic pH fluctuation of the tumor microenvironment, cells were reconditioned to neutral pH after experimental acidosis. This study assessed cell proliferation and viability by sulforhodamine B and flow cytometry. Individual and collective cell migration was analyzed by wound healing, time lapse, and transwell assays. Modifications of cell phenotype, EMT induction and stemness potential were investigated by qRT-PCR, western blot, and immunofluorescence. Finally, resistance to chemo- and radiotherapy of OSCC when exposed to acidified environmental conditions (pH 6.8) was determined. KEY FINDINGS: The exposure to an acidic microenvironment caused an initial reduction of OSCC cells viability, followed by an adaptation process. Acidic adapted cells acquired a mesenchymal-like phenotype along with increased migration and motility indexes. Moreover, tumoral extracellular acidity was capable to induce cellular stemness and to increase chemo- and radioresistance of oral cancer cells. SIGNIFICANCE: In summary, the results showed that the acidic microenvironment leads to a more aggressive and treatment resistant OSCC cell population.

Evaluation of the biological and physicochemical properties of calcium silicate-based and epoxy resin-based root canal sealers.

Biocompatibility, dimensional stability, radiopacity, flow, and low solubility are the characteristics of an ideal endodontic sealer. This study evaluated and compared in vivo and in vitro biological and physicochemical properties of calcium silicate-based sealers: Sealer Plus BC (BC), MTA Fillapex (MF); and resin-based sealers: AH Plus (AHP) and Sealer Plus (SP). Apical papilla cells were exposed to sealer extracts and subjected to MTT, SRB, scratch, alkaline phosphatase enzyme activity (ALP) and Alizarin red staining (ALZ) assays. Sealers were histologically evaluated in connective tissue of Wistar rats in different periods. Radiopacity, film thickness, flow, setting time, pH and element analyses were investigated. BC had better results compared to AHP and MF at hour 72 for MTT assay (p < .05), and the highest cell viability under SRB (p < .05). All sealers presented ALP activity. BC presented the highest mineralized deposition under ALZ (p < .05). BC and MF promoted wound healing. All sealers induced an initial inflammation reaction that decreased over time. Eosinophils were observed at day 7 in MF (p < .05). Despite MF did not present final setting time, the sealers properties were in accordance to ISO 6876/2012 and ASTM C266-08. All sealers presented cell viability and biocompatibility. BC presented higher pH values and bioactivity. The materials tested showed physico-chemical properties in accordance with standards, except for MF setting time.

Manifestações oftalmológicas em paciente com aneurisma gigante de carótida interna / Ophthalmological manifestation in patient with gigantic aneurysm of the internal carotid

RESUMO Os aneurismas intracranianos são dilatações em segmentos arteriais que irrigam o sistema nervoso central. Acometem 2% da população e as alterações oftalmológicas podem ser as primeiras manifestações do quadro. O objetivo deste relato foi descrever um caso de aneurisma de artéria carótida interna que cursou com restrição da movimentação ocular, alteração do reflexo fotomotor, ptose palpebral, dor facial e cervical. O diagnóstico foi confirmado pela identificação do aneurisma por meio do exame de angiografia cerebral. Foi realizado teste de oclusão por balão, cujo resultado positivo possibilitou a oclusão total da artéria carótida interna por meio de ligadura cirúrgica, procedimento este realizado com sucesso.

ABSTRACT Intracranial aneurysms are dilations in segments of the arteries that irrigate the central nervous system. They affect 2% of the population and the ophthalmologic disorders may be the first evidence in the clinical examination. The aim of the report is to describe a case of an internal carotid artery aneurysm that showed restrictions of ocular movements, change of pupillary light reflex, palpebral ptosis, facial, and cervical pain. This diagnosis was confirmed by the identification of the aneurysm through angiography. A balloon occlusion test was performed, and its positive result made a complete occlusion of the Internal Carotid Artery possible through surgery ligation, procedure that was successful.

Estrutura e dinâmica familiar de pessoas com deficiência intelectual / Family structure and dynamics of people with intellectual disabilities

O presente estudo teve o objetivo de explorar a estrutura e a dinâmica de famílias com um filho com deficiência intelectual. Foi utilizada a metodologia qualitativa, a partir do estudo de casos múltiplos, de cinco famílias residentes no Distrito Federal, com análise descritiva e exploratória de entrevistas semiestruturadas para a construção de genogramas, ecomapas, e o ciclo de vital das famílias. Os resultados apresentaram a estrutura e dinâmica familiar através dos dados sociodemográficos, das expectativas atuais e futuras, estratégias de enfrentamento e do ciclo vital. Sugere-se o conhecimento de estratégias familiares adequadas para a transição futura de cuidados no contexto da deficiência intelectual.

This study aimed at exploring the structure and dynamics of families with a child with intellectual disabilities. The qualitative methodology was used, based on the study of multiple cases, of five families living in the Federal District, with descriptive and exploratory analysis of semi-structured interviews for the construction of genograms, ecomapas, and the life cycle of the families. The results presented the family structure and dynamics through sociodemographic data, current and future expectations, coping strategies and life cycle. It is suggested the knowledge of adequate family strategies for the future transition of care in the context of intellectual disability.

Physicochemical and biological evaluation of a triazine-methacrylate monomer into a dental resin.

OBJECTIVES: This study aimed to (1) formulate blend resins with 2.5 or 5 wt.% of the methacrylate monomer 1,3,5-triacryloylhexahydro-1,3,5-triazine (TAT), and (2) to evaluate the blend resins regarding the physicochemical and biological properties. METHODS: The base resin was formulated mixing 60 wt.% of bisphenol A glycol dimethacrylate and 40 wt.% of triethylene glycol dimethacrylate with photoinitiator/co/initiator system. TAT was added at 2.5 (G2.5%) or 5 (G5%) wt.%, and a group without TAT was used as control (Gctrl). The resins were analyzed for degree of conversion (DC), Knoop hardness (KHN), softening in solvent (ΔKHN), ultimate tensile strength (UTS), contact angle, surface free energy (SFE), antibacterial activity against Streptococcus mutans biofilm formation, and cytotoxicity against human keratinocytes. RESULTS: There was no difference for the DC (p = 0.676). The addition of TAT at 5 wt.% induced higher KHN (p<0.001), higher resistance against softening in solvent (p<0.001), and higher UTS (p = 0.04). There were no statistically significant differences for contact angle with water (p = 0.106), α-bromonaphtalene (p = 0.454), and SFE (p = 0.172). The higher the TAT concentration, the higher the antibacterial activity (p<0.001). G2.5% showed no cytotoxicity compared to Gctrl (p>0.05), and G5% induced lower cell viability (p<0.05). CONCLUSIONS: The addition of 2.5 wt.% of TAT is suitable for conveying antibacterial activity for dental resins without changing the physicochemical properties or impairing the cytotoxic effect. CLINICAL RELEVANCE: Methacrylate monomers that decrease bacterial viability and copolymerize with the resin matrix are exciting approaches to developing therapeutic materials. TAT showed promising properties to may hamper and prevent carious lesions when incorporated into dental materials. Further evaluations with higher cariogenic challenges will be carried to analyze the formulated materials.

Lipid Coating of Chitosan Nanogels for Improved Colloidal Stability and In Vitro Biocompatibility.

Chitosan-based carriers have coined their position as delivery agents. When assembled with polyanions into nanogels (NG), these vectors have enabled the delivery of drugs, genes, and proteins to a myriad of applications. However, the chemical and colloidal instability of chitosan nanoformulations in physiologically compatible media prejudices in vitro biocompatibility and, thus, scale-up applications. To overcome this issue, we envisaged the coating of chitosan nanogel with phospholipids. In this investigation, we report a two-stage synthesis of hybrid lipid-coated chitosan nanogels, named nanolipogels (NLG), to improve colloidal stability and in vitro biocompatibility over chitosan NG. Practically, we employed a mixing platform to first prepare chitosan NG by ionic gelation, dilute the suspension, and, in a second stage, coat the NG with lipids. We demonstrate that lipid coating increased particle size and reversed the ζ-potential to negative values, suggesting the successful formation of NLG, while maintaining a homogeneous size distribution (PDI < 0.25). Furthermore, multiple light scattering analysis confirmed NLG improved colloidal stability in phosphate buffer saline and cell culture medium, with respect to NG. Finally, lipid coating completely abrogated the cytotoxicity of NG when incubated at 50 µg·mL-1 with HeLa, U87, or b.End3 cell lines and significantly improved the biocompatibility at 100 and 150 µg·mL-1. Future investigations will explore how the lipid coating affects drug loading, release profile, and the ability of NLG to deliver drugs and genes in vitro.