Monomer release, cell adhesion, and cell viability of indirect restorative materials manufactured with additive, subtractive, and conventional methods.

PURPOSE: The aim of this study was to measure residual monomer, cell adhesion, and cell viability of 3-dimensional printable permanent resin (PR), hybrid ceramic block (HCB), and indirect composite (IC) produced with additive, subtractive, and conventional techniques. METHODS: Five 8 × 8 × 2 mm3 samples of each material were prepared for each experiment. In a 24-h period, monomer release was analyzed with high-performance liquid chromatography, and cell viability and adhesion were evaluated with the water-soluble tetrazolium salt test. Data were analyzed with IBM SPSS Statistics 26.0 statistical software, and results were regarded as significant at α = 0.05. RESULTS: Monomer release (triethylene glycol dimethacrylate, urethane dimethacrylate, and Bisphenol A glycerolate dimethacrylate) was significantly higher in the IC group. Mean cell viability was significantly lower in the HCB group than in the IC group. CONCLUSION: All monomers in the tested materials were released at rates that were below clinical significance. Cell adhesion rates in the groups were similar. Cytotoxic response was classified as minor in the HCB and PR groups and non-cytotoxic in the IC group.

Evaluation of manual dexterity when using a mirror of dental students and academicians with increasing professional experience.

INTRODUCTION: To evaluate the manual dexterity of dentistry students and academicians when using a mirror according to increasing professional experience. MATERIALS AND METHODS: The study included 72 subjects, grouped according to professional experience 5th year dentistry students (DS) accepted as no experience-academicians with 1-4 years experience (A5L)-academicians with ≥5 years experience (A5M). Direct and indirect visualisation using the mirror was evaluated with the completion times of the O'Connor Finger Dexterity Test. RESULTS: The hand grip strength values of both left and right hand were found to be statistically significantly higher in all the males than in the females (p < .05). The indirect test times (ITT) using the mirror were significantly longer for males than for females (p = .001), and no significant difference was determined between the genders in the direct test times (DTT) (p > .05). For all the study participants, DTT shortened as professional experience increased (p < .05). In addition, the mean values of both DTT (p = .031) and ITT (p = .028) in the DS group were statistically significantly longer than the A5M group. CONCLUSION: With increasing professional experience, manual dexterity was determined to increase, and females were found to be more successful in the direct manual dexterity tests independently of the groups. The statistical significance between the DS and A5M groups, especially in the hand dexterity test with a mirror, shows the importance of experience. The study results demonstrated that professional manual dexterity in dentistry can be developed with increasing practical application.

Effect of thickness on the degree of conversion, monomer elution, depth of cure and cytotoxicity of bulk-fill composites.

PURPOSE: Today, bulk-fill composites are used as a single layer with a thickness of up to 4-5 mm. However, is proper polymerization achieved with this increased thickness? METHODS: This study was designed to investigate the effect of thickness on the degree of conversion (DC) (n = 6), the elution of monomers (n = 6), depth of cure (DoC) (n = 10) and cytotoxicity (n = 6) of the bulk-fill composites SDR Flow Plus (SDR), SonicFill2 SingleFill (SF) and ACTIVA Bioactive Restorative (ACT) in comparison to the conventional G-aenial Posterior (GC). Two-way analysis of variance (ANOVA) was used to assess the interaction between materials and surfaces, and one-way ANOVA and Tukey tests were used to compare the degree of conversion, monomer elution and cytotoxicity values (P < 0.05). RESULTS: The highest DC was found at the top surface of SDR, while the lowest DC was found at SF. The V2 mm/V0 mm DoC ratios of the composites except ACTs were appropriate according to the threshold. None of the composites were cytotoxic on day 1. CONCLUSION: In bulk-fill composites, DC decreased and monomer elution increased with increasing depth. The V4 mm/V0 mm ratios of all bulk-fill groups were not appropriate. Additionally, only ACTs had a cell viability of <70% on day 7.

Generation of two iPSC lines from long QT syndrome patients carrying SNTA1 variants.

Long QT syndrome (LQTS) is an inherited cardiovascular disorder characterized by electrical conduction abnormalities leading to arrhythmia, fainting, seizures, and an increased risk of sudden death. There are over 15 genes involved in causing LQTS, including SNTA1. Here we generated two human-induced pluripotent stem cell (iPSC) lines from two LQT patients carrying a missense mutation in SNTA1 (c.1088A > C). Both lines showed normal morphological properties, expressed pluripotency markers, showed a normal karyotype profile, and had the ability to differentiate into the three germ layers, making them a valuable tool to model LQTS to investigate the pathological mechanisms related to this SNTA1 variant.

Generation of two induced pluripotent stem cell lines from dilated cardiomyopathy patients caused by heterozygous mutations in the HCN4 gene.

Dilated cardiomyopathy (DCM) is a progressive heart muscle disease that can culminate with heart failure and death. Mutations in several genes can cause DCM, including hyperpolarization-activated cyclic nucleotide-gated channel (HCN4), which has a critical function in the autonomic control of the heart rate. Here, we generated two human induced pluripotent stem cell (iPSC) lines generated from two DCM patients carrying variants in the HCN4 gene (c.2587G > T and c.2846G > A). Both lines display normal karyotype, typical morphology of pluripotent stem cells, and differentiate into all three germ layers in vitro. These lines are valuable resources for studying the pathological mechanisms of DCM.

ER Stress-Induced Sphingosine-1-Phosphate Lyase Phosphorylation Potentiates the Mitochondrial Unfolded Protein Response.

The unfolded protein response (UPR) is an elaborate signaling network that evolved to maintain proteostasis in the endoplasmic reticulum (ER) and mitochondria (mt). These organelles are functionally and physically associated, and consequently, their stress responses are often intertwined. It is unclear how these two adaptive stress responses are coordinated during ER stress. The inositol-requiring enzyme-1 (IRE1), a central ER stress sensor and proximal regulator of the UPRER, harbors dual kinase and endoribonuclease (RNase) activities. IRE1 RNase activity initiates the transcriptional layer of the UPRER, but IRE1's kinase substrate(s) and their functions are largely unknown. Here, we discovered that sphingosine 1-phosphate (S1P) lyase (SPL), the enzyme that degrades S1P, is a substrate for the mammalian IRE1 kinase. Our data show that IRE1-dependent SPL phosphorylation inhibits SPL's enzymatic activity, resulting in increased intracellular S1P levels. S1P has previously been shown to induce the activation of mitochondrial UPR (UPRmt) in nematodes. We determined that IRE1 kinase-dependent S1P induction during ER stress potentiates UPRmt signaling in mammalian cells. Phosphorylation of eukaryotic translation initiation factor 2α (eif2α) is recognized as a critical molecular event for UPRmt activation in mammalian cells. Our data further demonstrate that inhibition of the IRE1-SPL axis abrogates the activation of two eif2α kinases, namely double-stranded RNA-activated protein kinase (PKR) and PKR-like ER kinase upon ER stress. These findings show that the IRE1-SPL axis plays a central role in coordinating the adaptive responses of ER and mitochondria to ER stress in mammalian cells.

PACT establishes a posttranscriptional brake on mitochondrial biogenesis by promoting the maturation of miR-181c.

The double-stranded RNA-dependent protein kinase activating protein (PACT), an RNA-binding protein that is part of the RNA-induced silencing complex, plays a key role in miR-mediated translational repression. Previous studies showed that PACT regulates the expression of various miRs, selects the miR strand to be loaded onto RNA-induced silencing complex, and determines proper miR length. Apart from PACT's role in mediating the antiviral response in immune cells, what PACT does in other cell types is unknown. Strikingly, it has also been shown that cold exposure leads to marked downregulation of PACT protein in mouse brown adipose tissue (BAT), where mitochondrial biogenesis and metabolism play a central role. Here, we show that PACT establishes a posttranscriptional brake on mitochondrial biogenesis (mitobiogenesis) by promoting the maturation of miR-181c, a key suppressor of mitobiogenesis that has been shown to target mitochondrial complex IV subunit I (Mtco1) and sirtuin 1 (Sirt1). Consistently, we found that a partial reduction in PACT expression is sufficient to enhance mitobiogenesis in brown adipocytes in culture as well as during BAT activation in mice. In conclusion, we demonstrate an unexpected role for PACT in the regulation of mitochondrial biogenesis and energetics in cells and BAT.

Intercepting IRE1 kinase-FMRP signaling prevents atherosclerosis progression.

Fragile X Mental Retardation protein (FMRP), widely known for its role in hereditary intellectual disability, is an RNA-binding protein (RBP) that controls translation of select mRNAs. We discovered that endoplasmic reticulum (ER) stress induces phosphorylation of FMRP on a site that is known to enhance translation inhibition of FMRP-bound mRNAs. We show ER stress-induced activation of Inositol requiring enzyme-1 (IRE1), an ER-resident stress-sensing kinase/endoribonuclease, leads to FMRP phosphorylation and to suppression of macrophage cholesterol efflux and apoptotic cell clearance (efferocytosis). Conversely, FMRP deficiency and pharmacological inhibition of IRE1 kinase activity enhances cholesterol efflux and efferocytosis, reducing atherosclerosis in mice. Our results provide mechanistic insights into how ER stress-induced IRE1 kinase activity contributes to macrophage cholesterol homeostasis and suggests IRE1 inhibition as a promising new way to counteract atherosclerosis.

S-adenosylmethionine inhibits la ribonucleoprotein domain family member 1 in murine liver and human liver cancer cells.

BACKGROUND AND AIMS: Methionine adenosyltransferase 1A (MAT1A) is responsible for S-adenosylmethionine (SAMe) biosynthesis in the liver. Mice lacking Mat1a have hepatic SAMe depletion and develop NASH and HCC spontaneously. Several kinases are activated in Mat1a knockout (KO) mice livers. However, characterizing the phospho-proteome and determining whether they contribute to liver pathology remain open for study. Our study aimed to provide this knowledge. APPROACH AND RESULTS: We performed phospho-proteomics in Mat1a KO mice livers with and without SAMe treatment to identify SAMe-dependent changes that may contribute to liver pathology. Our studies used Mat1a KO mice at different ages treated with and without SAMe, cell lines, in vitro translation and kinase assays, and human liver specimens. We found that the most striking change was hyperphosphorylation and increased content of La-related protein 1 (LARP1), which, in the unphosphorylated form, negatively regulates translation of 5'-terminal oligopyrimidine (TOP)-containing mRNAs. Consistently, multiple TOP proteins are induced in KO livers. Translation of TOP mRNAs ribosomal protein S3 and ribosomal protein L18 was enhanced by LARP1 overexpression in liver cancer cells. We identified LARP1-T449 as a SAMe-sensitive phospho-site of cyclin-dependent kinase 2 (CDK2). Knocking down CDK2 lowered LARP1 phosphorylation and prevented LARP1-overexpression-mediated increase in translation. LARP1-T449 phosphorylation induced global translation, cell growth, migration, invasion, and expression of oncogenic TOP-ribosomal proteins in HCC cells. LARP1 expression is increased in human NASH and HCC. CONCLUSIONS: Our results reveal a SAMe-sensitive mechanism of LARP1 phosphorylation that may be involved in the progression of NASH to HCC.

Are resin-containing pulp capping materials as reliable as traditional ones in terms of local and systemic biological effects?

The aim of this study was to compare the local and systemic effects of current pulp capping materials containing resin with those of traditional materials in an animal study. A total of 48 rats were used: a control group (n=12) (sub-control and negative control), a resin-containing group (n=18) (Calcimol LC, Theracal LC, Activa-BioActive Base/Liner), and a traditional group (n=18) (Biodentine, ProRoot MTA, Dycal). The materials which had been placed in polyethylene tubes were implanted in subcutaneous pockets. The rats were sacrificed at 1, 2, or 4 weeks. Evaluations were made of subcutaneous connective tissue, the left kidney, liver, and blood samples. Of all the study groups, MTA demonstrated biocompatibility at a level close to that of the control groups. Inflammation was observed to be more severe in resin-containing materials, but Activa Base/Liner showed a more successful local and systemic tissue response.

Inositol-requiring enzyme-1 regulates phosphoinositide signaling lipids and macrophage growth.

The ER-bound kinase/endoribonuclease (RNase), inositol-requiring enzyme-1 (IRE1), regulates the phylogenetically most conserved arm of the unfolded protein response (UPR). However, the complex biology and pathology regulated by mammalian IRE1 cannot be fully explained by IRE1's one known, specific RNA target, X box-binding protein-1 (XBP1) or the RNA substrates of IRE1-dependent RNA degradation (RIDD) activity. Investigating other specific substrates of IRE1 kinase and RNase activities may illuminate how it performs these diverse functions in mammalian cells. We report that macrophage IRE1 plays an unprecedented role in regulating phosphatidylinositide-derived signaling lipid metabolites and has profound impact on the downstream signaling mediated by the mammalian target of rapamycin (mTOR). This cross-talk between UPR and mTOR pathways occurs through the unconventional maturation of microRNA (miR) 2137 by IRE1's RNase activity. Furthermore, phosphatidylinositol (3,4,5) phosphate (PI(3,4,5)P3 ) 5-phosphatase-2 (INPPL1) is a direct target of miR-2137, which controls PI(3,4,5)P3 levels in macrophages. The modulation of cellular PI(3,4,5)P3 /PIP2 ratio and anabolic mTOR signaling by the IRE1-induced miR-2137 demonstrates how the ER can provide a critical input into cell growth decisions.

Double bond configuration of palmitoleate is critical for atheroprotection.

OBJECTIVE: Saturated and trans fat consumption is associated with increased cardiovascular disease (CVD) risk. Current dietary guidelines recommend low fat and significantly reduced trans fat intake. Full fat dairy can worsen dyslipidemia, but recent epidemiological studies show full-fat dairy consumption may reduce diabetes and CVD risk. This dairy paradox prompted a reassessment of the dietary guidelines. The beneficial metabolic effects in dairy have been claimed for a ruminant-derived, trans fatty acid, trans-C16:1n-7 or trans-palmitoleate (trans-PAO). A close relative, cis-PAO, is produced by de novo lipogenesis and mediates inter-organ crosstalk, improving insulin-sensitivity and alleviating atherosclerosis in mice. These findings suggest trans-PAO may be a useful substitute for full fat dairy, but a metabolic function for trans-PAO has not been shown to date. METHODS: Using lipidomics, we directly investigated trans-PAO's impact on plasma and tissue lipid profiles in a hypercholesterolemic atherosclerosis mouse model. Furthermore, we investigated trans-PAO's impact on hyperlipidemia-induced inflammation and atherosclerosis progression in these mice. RESULTS: Oral trans-PAO supplementation led to significant incorporation of trans-PAO into major lipid species in plasma and tissues. Unlike cis-PAO, however, trans-PAO did not prevent organelle stress and inflammation in macrophages or atherosclerosis progression in mice. CONCLUSIONS: A significant, inverse correlation between circulating trans-PAO levels and diabetes incidence and cardiovascular mortality has been reported. Our findings show that trans-PAO can incorporate efficiently into the same pools that its cis counterpart is known to incorporate into. However, we found trans-PAO's anti-inflammatory and anti-atherosclerotic effects are muted due to its different structure from cis-PAO.

Qualitative and Quantitative Evaluation of Cytotoxicity of Five Different One-Step Self-Etching Adhesives.

PURPOSE: To qualitatively and quantitatively compare the cytotoxic potentials of five different one-step self-etching adhesives: Prime&Bond One-Select (PB-OS), Optibond All-in-One (OB-AIO), G-Bond (GB), Clearfil Universal Bond (CUB), Single Bond Universal (SBU). MATERIALS AND METHODS: During the first stage of the study, the cytotoxic activities of the test materials were evaluated qualitatively using the direct contact method. In this method, the test materials were placed directly into a monkey kidney epithelial cell culture medium. Reaction zones which occurred in the culture medium were evaluated, in addition to the density and changes in the morphology of the cells. During the second stage, the cytotoxic potential of four different dilutions (1%, 0.1%, 0.01%, 0.001%) of the test materials on L929 rat fibroblast cells was quantitatively evaluated at three different time periods (24 h, 48 h, 72 h) with the MTT tetrazolium-based assay. RESULTS: In the first stage, a zone exceeding 1 cm was observed around or below SBU, CUB, GB and OB-AIO. In PB-OS, the zone borders were approximately 1 cm. In the second stage after the MTT assay, CUB was the most cytotoxic after 24 h, GB and SBU after 48 h, and OB-AIO after 72 h. CONCLUSION: All adhesives tested showed different degrees of cytotoxicity, which statistically significantly increased with dose. Changes were seen related to time.

[Comparing the occurrence of Trichomonas vaginalis infections today to ten years ago among women prostitutes and gynecology and obstetrics patients in Istanbul]. / Istanbul'da Kadin Hastaliklari Hastalari ile Hayat Kadinlarinda Trichomonas vaginalis Görülme Sikliginin 10 Yil Önceki Oranla Karsilastirilmasi

OBJECTIVE: Trichomonas vaginalis is is a monoxenous parasite which lives in human urogenital systems and causes sex transmitted disease through human sexual contact. Disease frequency has been seen at different rates in different communities or in the same community depending on people's sociocultural status. Previously we made a study for determining prevalence of T. vaginalis infection in woman living in Istanbul. We made this present study for determining any difference in prevalence in comparison to the results of ten years earlier. METHODS: A total number of 207 vaginal discharge samples which were collected from two different hospitals, (93 from Venereal Diseases Hospital [VDH] and 114 from Cerrahpasa Ob&Gyn Clinic), were evaluated under direct microscopy and were cultured for T. vaginalis in a Cystein-Peptone-Liver-Maltose (CPLM) medium. RESULTS: T. vaginalis was observed under direct microscopy and grew in culture in 2 (0.97%) of 207 vaginal discharge samples [1 (1.1%) patient from VDH and 1 (0.9%) patient from Cerrahpasa]. CONCLUSION: The incidence of trichomoniasis has significantly decreased compared to the year 2000 in both VDH and Cerrahpasa populations (p=0.038) according to X2 test results. This epidemiologic study shows the importance of social development in the incidence of infectious diseases.