Porphyromonas gingivalis activates Heat-Shock-Protein 27 to drive a LC3C-specific probacterial form of select autophagy that is redox sensitive for intracellular bacterial survival in human gingival mucosa.

Porphyromonas gingivalis , a major oral pathobiont, evades canonical host pathogen clearance in human primary gingival epithelial cells (GECs) by initiating a non-canonical variant of autophagy consisting of Microtubule-associated protein 1A/1B-light chain 3 (LC3)-rich autophagosomes, which then act as replicative niches. Simultaneously, P. gingivalis inhibits apoptosis and oxidative-stress, including extracellular-ATP (eATP)-mediated reactive-oxygen-species (ROS) production via phosphorylating Heat Shock Protein 27 (HSp27) with the bacterial nucleoside-diphosphate-kinase (Ndk). Here, we have mechanistically identified that P. gingivalis -mediated induction of HSp27 is crucial for the recruitment of the LC3 isoform, LC3C, to drive the formation of live P. gingivalis -containing Beclin1-ATG14-rich autophagosomes that are redox sensitive and non-degrading. HSp27 depletions of both infected GECs and gingiva-mimicking organotypic-culture systems resulted in the collapse of P. gingivalis -mediated autophagosomes, and abolished P. gingivalis -induced LC3C-specific autophagic-flux in a HSp27-dependent manner. Concurrently, HSp27 depletion accompanied by eATP treatment abrogated protracted Beclin 1-ATG14 partnering and decreased live intracellular P. gingivalis levels. These events were only partially restored via treatments with the antioxidant N-acetyl cysteine (NAC), which rescued the cellular redox environment independent of HSp27. Moreover, the temporal phosphorylation of HSp27 by the bacterial Ndk results in HSp27 tightly partnering with LC3C, hindering LC3C canonical cleavage, extending Beclin 1-ATG14 association, and halting canonical maturation. These findings pinpoint how HSp27 pleiotropically serves as a major platform-molecule, redox regulator, and stepwise modulator of LC3C during P. gingivalis -mediated non-canonical autophagy. Thus, our findings can determine specific molecular strategies for interfering with the host-adapted P. gingivalis ' successful mucosal colonization and oral dysbiosis.

Opportunistic pathogen Porphyromonas gingivalis targets the LC3B-ceramide complex and mediates lethal mitophagy resistance in oral tumors.

Mechanisms by which Porphyromonas gingivalis (P. gingivalis) infection enhances oral tumor growth or resistance to cell death remain elusive. Here, we determined that P. gingivalis infection mediates therapeutic resistance via inhibiting lethal mitophagy in cancer cells and tumors. Mechanistically, P. gingivalis targets the LC3B-ceramide complex by associating with LC3B via bacterial major fimbriae (FimA) protein, preventing ceramide-dependent mitophagy in response to various therapeutic agents. Moreover, ceramide-mediated mitophagy is induced by Annexin A2 (ANXA2)-ceramide association involving the E142 residue of ANXA2. Inhibition of ANXA2-ceramide-LC3B complex formation by wild-type P. gingivalis prevented ceramide-dependent mitophagy. Moreover, a FimA-deletion mutant P. gingivalis variant had no inhibitory effects on ceramide-dependent mitophagy. Further, 16S rRNA sequencing of oral tumors indicated that P. gingivalis infection altered the microbiome of the tumor macroenvironment in response to ceramide analog treatment in mice. Thus, these data provide a mechanism describing the pro-survival roles of P. gingivalis in oral tumors.

Sprint and Anaerobic Power with the Soccer-Specific ACTN3 Gene: A Distintive Example / Sprint y Potencia Anaeróbica con el Gen ACTN3 Específico del Fútbol: Un Ejemplo Distintivo

SUMMARY: The aim of this study is twofold: (1) to identify differences in certain anaerobic parameters (10m sprint, 30m sprint, anaerobic power, and Illinois agility tests) between professional and amateur soccer players, and (2) to determine whether there is a difference in the ACTN3 gene polymorphism between professional and amateur soccer players. Ultimately, the goal is to reveal which parameters contribute to the differentiation in these two aspects. A total of 133 volunteer soccer players, including 71 professionals and 62 amateurs, participated in the research. DNA extraction from buccal epithelial cells was performed using a commercial kit to determine the genetic background of the athletes, and Real-Time PCR was conducted for genotyping. Statistical analysis of the findings obtained from the test results was performed using the SPSS 23 (SPSS Inc., Chicago, IL, USA) package program. The homogeneity of variance of the data was assessed using the Levene Test, and normal distribution analyses were conducted using the Shapiro-Wilk Test. Chi-square and Mann-Whitney U tests were employed for parameter analysis. The significance level was set at p0.05). However, there is a statistically significant difference in anaerobic parameters (10m sprint, 30m sprint, and anaerobic power) except for the Illinois test (p<0.05). In conclusion, our study found that gene polymorphism is not a differentiating factor between professional and amateur soccer players, but speed (10m and 30m) and anaerobic power parameters are differentiating factors.

Los objetivos de este estudio fueron: 1º identificar diferencias en ciertos parámetros anaeróbicos (sprint de 10 m, sprint de 30 m, potencia anaeróbica y pruebas de agilidad de Illinois) entre jugadores de fútbol profesionales y amateurs, y 2º determinar si existe una diferencia en el polimorfismo del gen ACTN3 entre jugadores de fútbol profesionales y aficionados. En definitiva, el objetivo fue revelar qué parámetros contribuyen a la diferenciación en estos dos aspectos. En la investigación participaron un total de 133 jugadores de fútbol voluntarios, incluidos 71 profesionales y 62 aficionados. La extracción de ADN de las células epiteliales orales se realizó utilizando un kit comercial para determinar los antecedentes genéticos de los atletas y se realizó una PCR en tiempo real para el genotipado. El análisis estadístico de los hallazgos obtenidos a partir de los resultados de las pruebas se realizó utilizando el programa de paquete SPSS 23 (SPSS Inc., Chicago, IL, EE. UU.). La homogeneidad de la varianza de los datos se evaluó mediante la prueba de Levene y los análisis de distribución normal se realizaron mediante la prueba de Shapiro-Wilk. Para el análisis de parámetros se emplearon las pruebas de Chi-cuadrado y U de Mann-Whitney. El nivel de significancia se fijó en p0,05). Sin embargo, existe una diferencia estadísticamente significativa en los parámetros anaeróbicos (sprint de 10 m, sprint de 30 m y potencia anaeróbica) excepto para la prueba de Illinois (p<0,05). En conclusión, nuestro estudio encontró que el polimorfismo genético no es un fac- tor diferenciador entre jugadores de fútbol profesionales y amateurs, pero sí los parámetros de velocidad (10 m y 30 m) y potencia anaeróbica.

The evaluation of L-arginine solution as a solvent for propolis extraction: The phenolic profile, antioxidant, antibacterial activity, and in vitro bioaccessibility.

Ethanol has been widely used for the extraction of propolis. Due to its certain disadvantages, there has been an ongoing search to find alternative non-ethanolic extraction solvents. This study aimed to compare the phenolics, antioxidant, and antibacterial activity of propolis extracts prepared with 70% ethanol (EWE), propylene glycol (PGE), and L-arginine solution (BE). All extracts were subjected to an in vitro simulated digestion procedure, and the phenolic profile of non-digested and digested samples was determined by using LC-MS/MS. Additionally, the change in total phenolic (TPC), total flavonoid content (TFC), and antioxidant capacities were determined at each digestion phase. TPC and TFC of non-digested propolis extracts had similar values, although BE showed higher antioxidant capacity (p < .05). The amount of TPC reached or transformed at the intestinal stage was higher for BE and PG compared to EWE. BE also provided the highest antioxidant capacity assay in digested samples. The most common phenolics were pinocembrin, pinobanskin, galangin, and CAPE in non-digested extracts. However, their concentration was drastically reduced by digestion, and their recovery (R%) ranged from 0% to 9.38% of the initial amount detected in the non-digested extracts. Chrysin was the most bioaccessible flavonoid in all extracts. Among phenolic acids, the highest R% was determined for trans-cinnamic acid (22.14%) from BE. All extracts showed in vitro inhibitory activity against Escherichia coli and Staphylococcus aureus. This study suggests that an L-arginine solution could be used as an alternative solvent to ethanol and propylene glycol for propolis extraction.

Use of polyglycolic acid-hyaluronic acid/ß-tricalcium phosphate scaffold with or without mesenchymal stem cells found ineffective in treating osteochondral lesions in rabbit knees.

OBJECTIVE: In this experimental animal study, a novel bilayered scaffold used in the treatment of osteochondral defects in rabbit knees was evaluated. This novel scaffold's upper (cartilage) layer consists of polyglycolic acid and hyaluronic acid, and the lower (bone) layer consists of ß-tricalcium phosphate. The purpose of this study was to evaluate the efficacy of this novel scaffold, combined with or without mesenchymal stem cells (MSCs), in the treatment of osteochondral defects in rabbit knees. METHODS: Osteochondral defects were created in the left femoral trochlea of 30 rabbits. In group A, defects were treated with scaffold combined with MSCs; in group B, defects were treated with cell-free scaffolds; and group C was a control group with defects left untreated. In the 12th week, animals were sacrificed for macroscopic evaluation. RESULTS: The mean International Cartilage Repair Society (ICRS) macroscopic scores were 4.95 for group A, 6.16 for group B, and 8.25 for group C. The mean Oswestry Arthroscopic Scores (OAS) were 1.65 for group A, 3.39 for group B, and 6.05 for group C. The macroscopic scores were significantly higher in group C than group A for ICRS scores and group A and group B for OAS (P < .001, P < .000, P < .022). CONCLUSION: In essence, our findings indicate that the newly developed osteochondral scaffold, when tested in a rabbit model, is not as effective as expected in repairing full-thickness osteochondral defects, with or without the supplementation of MSCs. Further investigation is required to enhance the effectiveness of this novel combination.

The Distinguishing Factor in Soccer Players is Aerobic Performance, not the ACTN3 Gene / El Factor Distintivo en los Jugadores de Fútbol es el Rendimiento Aeróbico y no el gen ACTN3

SUMMARY: The purpose of this study was to reveal the differences between ACTN3 genotype (RR, RX, XX) and aerobic performance [Yo-Yo IRT1 (m), VO2 max (ml/kg/min)] in professional and regional amateur league soccer players and to reveal which of these parameters was a distinctive factor in these athletes.71 professional soccer players (age: 23.66 ± 4.11 years; body height: 1.79 ± 6.99 m; body weight: 76.02 ± 6.76 kg; body fat: 11.59±3.11 %) and 62 regional amateur soccer players (age: 23.63 ±3.77 years; body height: 1.81 ± 5.77 m; body weight: 76.36 ± 7.53 kg; body fat: 15.60±4.65 %) volunteered for the study. After DNA extraction from buccal epithelial cells via a commercial kit was performed for the genetic background of the athletes, Real-Time PCR was carried out for genotyping. Furthermore, Yo-Yo IRT1 test was performed to determine the aerobic performance of the soccer players. SPSS 23 (SPSS Inc., Chicago, IL, USA) package program was used for the statistical analysis of the data obtained in the tests. Shapiro-Wilk test for normality and Levene's test for homogeneity of variance were performed. Chi-Square, Independent Sample T Test and One Way ANOVA test were used in the analysis of the parameters. Statistical significance was set as p0.05); however, there was a statistical significance in favor of professional soccer players in terms of aerobic parameters (p<0.05). Consequently, it can be said that aerobic performance is the distinguishing factor, not the ACTN3 gene, in soccer players.

El objetivo de este estudio fue revelar las diferencias entre el genotipo ACTN3 (RR, RX, XX) y el rendimiento aeróbico [Yo-Yo IRT1 (m), VO2 max (ml/kg/min)] en jugadores de fútbol de ligas profesionales y amateurs regionales y determinar cuál de estos parámetros es un factor distintivo en estos deportistas. 71 futbolistas profesionales (edad: 23,66 ±4,11 años; altura corporal: 1,79 ± 6,99 m; peso corporal: 76,02 ± 6,76 kg; grasa corporal: 11,59±3,11 %) y 62 jugadores de fútbol amateur regionales (edad: 23,63 ± 3,77 años; altura corporal: 1,81 ± 5,77 m; peso corporal: 76,36 ± 7,53 kg; grasa corporal: 15,60 ± 4,65 %) se ofrecieron como voluntarios para el estudio. Después de realizar la extracción de ADN de las células epiteliales orales mediante un kit comercial para obtener los antecedentes genéticos de los atletas, se llevó a cabo una PCR en tiempo real para el genotipado. Además, se realizó la prueba Yo-Yo IRT1 para determinar el rendimiento aeróbico de los futbolistas. Para el análisis estadístico de los datos obtenidos en las pruebas se utilizó el programa SPSS 23 (SPSS Inc., Chicago, IL, EE. UU.). Se realizó la prueba de normalidad de Shapiro- Wilk y la prueba de homogeneidad de la varianza de Levene. En el análisis de los parámetros se utilizaron Chi-cuadrado, prueba T para muestra independiente y prueba ANOVA unidireccional. La significancia estadística se estableció en p0,05); sin embargo, hubo significación estadística a favor de los futbolistas profesionales en cuanto a los parámetros aeróbicos (p<0,05). En consecuencia, se puede decir que el rendimiento aeróbico es el factor distintivo, no el gen ACTN3, en los jugadores de fútbol.

Statins Modulate Microenvironmental Cues Driving Macrophage Polarization in Simulated Periodontal Inflammation.

Periodontal disease (PD) is a chronic inflammatory disorder characterized by the destruction of connective tissue, tooth loss, and systemic infections. Clinically, treatment of PD includes control of the etiologic factors via several modalities: initial therapy including scaling and root planing (SRP), corrective phase of surgical treatment, both with and without adjunct antimicrobial/pharmacological agents, followed by a maintenance/supportive periodontal therapy phase. Each treatment phase aims to control oral biofilm by addressing risk factors and etiology. Monotherapy of systemic antibiotics is insufficient compared to their use as an adjunct to SRP. The critical issue of systemic antimicrobial usage includes adverse patient outcomes and increased bacterial resistance. Therefore, alternative adjuncts to periodontal therapy have been sought. Statins are widely prescribed for the treatment of hypercholesterolemia and cardiovascular disease. Statins have demonstrated anti-inflammatory properties and immunomodulatory effects, and a few retrospective studies showed that statin patients exhibit fewer signs of periodontal inflammation than subjects without the medication. Despite the available clinical studies on the local administration of statins for PD, no studies have reported the macrophage polarization response. We have developed a gingival fibroblast-macrophage co-culture model to track macrophage response when exposed to a battery of microenvironmental cues mimicking macrophage polarization/depolarization observed in vivo. Using our model, we demonstrate that simvastatin suppresses macrophage inflammatory response and upregulates tissue homeostasis and M2 macrophage markers. Our findings support the usage of statins to mitigate periodontal inflammation as a valid strategy.

Rottlerin and genistein inhibit neuroblastoma cell proliferation and invasion through EF2K suppression and related protein pathways.

Neuroblastoma is one of the most common solid tumors in children younger than 1 year of age, with poor prognosis and survival rates. Therefore, novel molecular targets and therapeutic strategies are needed to prolong patient survival. For this purpose, we investigated the effects of rottlerin and genistein separately and in combination on neuroblastoma cells (SH-SY5Y, Kelly). First, the effects of rottlerin and genistein were investigated on cell proliferation. Different rottlerin (1-50 µM) and genistein (5-150 µM) doses were used as experimental groups compared to the control (DMSO/vehicle). The IC50 dose was found to be 5 µM for rottlerin and 30 µM for genistein (P < 0.0001). Other analyses, such as colony formation assays, annexin V/propidium iodide staining, matrigel invasion assays, and Western blot analysis, were performed with these doses and their combinations. To assess statistical significance, statistical analysis was conducted using the one-way ANOVA with the post hoc Tukey test. Our results showed that IC50 doses of rottlerin and genistein induced a significant reduction in cell proliferation, colony formation, and invasion in neuroblastoma cells (P < 0.0001). The combination of these doses increased the levels of inhibition of cell proliferation and invasion while decreasing the level of apoptosis (P 0.0001). Furthermore, these agents caused G1-cell cycle arrest in these cells. Our western blot data showed that rottlerin and genistein treatments markedly inhibit elongation factor 2 kinase (EF2K) and other pro-tumorigenic, metastatic proteins in neuroblastoma cells. These agents probably showed their anti-proliferative, anti-metastatic, and pro-apoptotic effects through EF2K downregulation. Our results suggested that rottlerin and genistein have inhibitory effects on cancer cell proliferation, invasion, and cell cycle and induce apoptosis in both cell lines. Combined treatment with rottlerin and genistein may be a viable approach and beneficial to neuroblastoma patients as the combined effect significantly suppresses the above-mentioned pathways.

Inhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Enhanced Alveolar Bone Regeneration by Promoting Angiogenesis.

Sphingosine-1-phosphate receptor 2 (S1PR2) is a G protein-coupled receptor that regulates various immune responses. Herein, we report the effects of a S1PR2 antagonist (JTE013) on bone regeneration. Murine bone marrow stromal cells (BMSCs) were treated with dimethylsulfoxide (DMSO) or JTE013 with or without infection by an oral bacterial pathogen Aggregatibacter actinomycetemcomitans. Treatment with JTE013 enhanced vascular endothelial growth factor A (VEGFA), platelet derived growth factor subunit A (PDGFA), and growth differentiation factor 15 (GDF15) gene expression and increased transforming growth factor beta (TGFß)/Smad and Akt signaling. Eight-week-old male C57BL/6J mice were challenged with ligatures around the left maxillary 2nd molar for 15 days to induce inflammatory bone loss. After ligature removal, mice were treated with diluted DMSO or JTE013 in the periodontal tissues 3 times per week for 3 weeks. Calcein was also injected twice to measure bone regeneration. Micro-CT scanning of maxillary bone tissues and calcein imaging revealed that treatment with JTE013 enhanced alveolar bone regeneration. JTE013 also increased VEGFA, PDGFA, osteocalcin, and osterix gene expressions in the periodontal tissues compared to control. Histological examination of periodontal tissues revealed that JTE013 promoted angiogenesis in the periodontal tissues compared to control. Our findings support that inhibition of S1PR2 by JTE013 increased TGFß/Smad and Akt signaling; enhanced VEGFA, PDGFA, and GDF15 gene expression; and subsequently promoted angiogenesis and alveolar bone regeneration.

Shared Biological Pathways and Processes in Patients with Intellectual Disability: A Multicenter Study.

BACKGROUND: Although the underlying genetic causes of intellectual disability (ID) continue to be rapidly identified, the biological pathways and processes that could be targets for a potential molecular therapy are not yet known. This study aimed to identify ID-related shared pathways and processes utilizing enrichment analyses. METHODS: In this multicenter study, causative genes of patients with ID were used as input for Disease Ontology (DO), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes enrichment analysis. RESULTS: Genetic test results of 720 patients from 27 centers were obtained. Patients with chromosomal deletion/duplication, non-ID genes, novel genes, and results with changes in more than one gene were excluded. A total of 558 patients with 341 different causative genes were included in the study. Pathway-based enrichment analysis of the ID-related genes via ClusterProfiler revealed 18 shared pathways, with lysine degradation and nicotine addiction being the most common. The most common of the 25 overrepresented DO terms was ID. The most frequently overrepresented GO biological process, cellular component, and molecular function terms were regulation of membrane potential, ion channel complex, and voltage-gated ion channel activity/voltage-gated channel activity, respectively. CONCLUSION: Lysine degradation, nicotine addiction, and thyroid hormone signaling pathways are well-suited to be research areas for the discovery of new targeted therapies in ID patients.

The importance microRNAs as a biomarker in lung cancer.

INTRODUCTION: Lung cancer (LC) is the most common cancer  in the world.Well known  causes are  long term  smoking, environmental influences and genetic variations. LC  is divided into two main types based on their histological phenotypes; small cell lung cancer (SCLC), and non-small cell lung cancer (NSCLC). The high specificity of these new screening methods, which are non-invasive, safe, inexpensive and simple to perform, is important in the early diagnosis and prognosis of cancer. MicroRNAs are  significant biomarkers on the diagnosis metastasis and targeted therapies of NSCLC. In our study, we aimed to investigate the potential of using microRNAs as a biomarker in the early diagnosis of lung cancer. MATERIAL AND METHOD: Twenty patients diagnosed with lung cancer and  twenty healthy individuals of the same age and gender were selected as the control group.  Sixteen microRNAs were studied from blood samples. RESULT: Sixteen miRNAs (Let -7c, Let-7g, miR-1, miR-21, miR-29a, miR-31, miR-34a, miR 103a, miR-141, miR-155, miR-193b, miR-200b, miR-205, miR-340, miR-486, miR-708) were selected for tests and MiR 181 and miR 192 were used as the endogenous control group in line with their binding potentials and gene expression levels. The most specific and sensitive miRNAs were mirR-29a, miR-103a, and miR486 according to endogen controls in patients and healthy subjects. DISCUSSION: A meta-analysis study showed that circulating miRNAs could be promising biomarkers for early diagnosis of lung cancer. Overall, 17 studies were included evaluating 35 miRNA markers and 19 miRNA panels in serum or plasma. The potential role of circulating miRNAs for non-invasive lung screening has been highlighted. In conclusion, there is a need for further validation studies for the use of three  miRNAs as a biomarker in the early diagnosis and prognosis of lung cancer.

Epigallocatechin-3-gallate and resveratrol attenuate hydrogen peroxide induced damage in neuronal cells.

PURPOSE: The purpose of this study is to investigate how the antioxidants resveratrol and epigallocatechin-3-gallate (EGCG) protect SH-SY5Y cells against damage caused by hydrogen peroxide (H2O2). METHODS: SH-SY5Y cells were pretreated with EGCG and resveratrol at concentrations of 0.1 µM, 1 µM, and 10 µM, individually and in various combinations. Cells were exposed to 250 µM H2O2 for 1-hour following a 24-hour pretreatment. The effects of EGCG and resveratrol on cellular survival against hydrogen peroxide toxicity were evaluated with the MTS. Caspase 3 levels were measured with caspase 3 ELISA test for evaluating survival. The clonogenic test was utilized to assess the colony forming capacity. RESULTS: The MTS test revealed that pretreatment of SH-SY5Y cells for 24 hours with EGCG and Resveratrol enhanced cellular survival against hydrogen peroxide damage at all dosages (p < 0.005). The caspase 3 ELISA test revealed that EGCG and resveratrol statistically substantially decreased caspase 3 levels and improved cellular survival via the caspase 3 pathway (p < 0.005). The clonogenic test findings show that resveratrol and EGCG statistically boost SH-SY5Y cells' potential to form colonies (p<0.005). CONCLUSIONS: By reducing caspase 3 levels in exposure to hydrogen peroxide damage, EGCG and resveratrol promote cellular viability (Tab. 1, Fig. 3, Ref. 37). Text in PDF www.elis.sk Keywords: epigallocatechin-3-gallate, resveratrol, hydrogen peroxide, neurodegeneration.

Comparison of postoperative pain after the use of different nickel titanium instrumentation systems: a randomized clinical trial.

Purpose: Postoperative pain is a common complication in endodontics contributed by multiple etiological factors, which consist canal preparation instruments and kinematics. The aim of this randomized clinical trial compare the postoperative pain in terms of intensity and incidence after the use of different nickel titanium (NiTi) file systems. Patients and methods: In this randomized clinical trial (NCT03791762), a total of 150 patients were root canal treated by 2 experienced endodontists according to a standardised protocol. The subjects were randomly assigned to 1 of the 3 groups according to preparation instrument used: ProTaper Next (Dentsply Sirona, Ballaigues, Switzerland), Reciproc Blue (VDW, Munich, Germany) and WaveOne Gold (Dentsply Sirona). Following preparation the teeth underwent standardized root canal treatment procedures in a single visit. The patients were contacted to gather information about the incidence of pain and intensity at 6th, 12th, 18th, 24th, 48th, and 72nd hours postoperatively. The data were analysed using chi-square, one-way analysis of variance and post hoc Tukey tests and logistic regression analysis with 5% significance threshold. Results: No significant difference was found among preparation groups in relation to the intensity of postoperative pain. The incidence of postoperative pain was significantly linked with the preoperative pain presence with odds ratio values ranging between 2.06 and 4.08 irrespective of the preparation technique (P<0.05). Conclusion: The effects of reciprocating and the continuous rotary systems on the intensity and incidence of postoperative pain were found to be similar.

Use of MicroRNAs as biomarkers in the early diagnosis of prostate cancer.

INTRODUCTION: Prostate cancer (PCa) is a common type of cancer in western countries and prominent cause of mortality in men. The aim of the study was to analyze circulating miRNAs as biomarkers in the sera of healthy individuals and prostate cancer cases without biopsy. MATERIAL AND METHODS: Twenty prostate cases, age (mean and range) 61,4±12.1 (45-73), and twenty healthy men, age 59,3±11.2 (44-70) were included to the study. The mean and range of prostate spesific antigen (PSA) in cancer cases and healthy individuals were 6.79±2.84 ng/ml (2.25-14.7) and 3.8±2.2 ng/ml (1.3-7.8) respectively. RESULTS: Seven miRNAs including two internal controls (Let7c, miR125b, miR141, miR145, miR 155, miR181 ve miR192) were evaluated in two groups. The level of miR141 was significantly lower in PCa cases than healthy individuals (p=0,004), and miR155 was significantly higher (p=0,005) in PCa cases. Both miRNAs were explored sensitive and spesific in the ROC analysis. Tumor mass were found to be associated with the level of miR-125b and miR-145. Conclusion; validation studies are required in wider patient groups in the subject of tumor effect and miRNA biomarkers in prostate cancer.

Method validation, residue and risk assessment of 260 pesticides in some leafy vegetables using liquid chromatography coupled to tandem mass spectrometry.

In the present study, QuEChERS method was optimized and validated for determination of 260 pesticides by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The method had a suitable linearity (R2 ≥ 0.99). Limit of detection (LOD) and limit of quantification (LOQ) were ranged from 0.56 to 2.99 µg kg-1 and 1.88 to 9.99 µg kg-1 respectively. Average recoveries varied from 71.14% to 118.83%. Repeatability conditions and intra-laboratory reproducibility conditions of the method expressed as %RSD were found between 0.55% and 16.69% and 0.74% to 19.06%, respectively. The method was used to detect pesticide residue levels of leafy vegetables collected from the open markets, greenhouses and wholesale vegetable market. The pesticide residues were detected in 57.6% of the all samples. In five samples, pesticide residues were above Maximum Residue Levels (MRLs). The risk assessment study for detected pesticides in samples indicated that leafy vegetables consumption was safe for consumers.

Inflammasome Activation in Gingival Epithelial Cells.

Inflammasomes are multiprotein complexes that assemble in host cells upon recognition of infection or danger via pattern recognition receptors and/or danger recognition receptors. The assembly of inflammasomes results in the activation of caspase-1 and is followed by the enzymatic maturation and secretion of inflammatory cytokines like interleukin 1ß (IL-1ß) and IL-18.In the oral cavity, gingival epithelial cells (GECs) line the mucosa and have a barrier role for invading pathogens. In these cells, the NLRP3 inflammasome is the best studied and has been shown to play a role in the inflammatory immune response against a variety of oral pathogens. As such, in order to study gingivitis and other oral pathologic inflammatory conditions, studying the activation of inflammasomes is important. The simplest way to detect inflammasome activation is to detect the activated caspase-1, as well as the secretion of mature IL-1ß and IL-18, via ELISA, Western blot, and immunofluorescence techniques.Here we describe the detection of NLRP3 inflammasome activation by the oral pathogen Porphyromonas gingivalis in human GECs via these three methods and mention other methods and techniques that we have successfully used together with these in our quest to understand the host-pathogen interaction.

Physical Growth of Patients with Hereditary Tyrosinaemia Type I: A Single-Centre Retrospective Study.

In a retrospective review, we aimed to assess long-term growth in 17 patients (n = 11 males) with hereditary tyrosinaemia type I (HTI). Median age at assessment was 15.6 years (5.7-26.6 years) and median age at diagnosis was 1 month (range: 0-16 months), with 35% (n = 6/17) symptomatic on presentation. From the age of 8 years, there was a noticeable change in median height, weight, and body-mass-index [BMI]-z-scores. Median height-for-age z-scores were consistently ≤ -1 (IQR -1.6, -0.5) during the first 8 years of life but increased with age. Weight-for-age z-scores ranged between -1 to 0 (IQR -1.2, 0.1) in the first 8 years; then increased to > 0.5 (IQR -0.3, 1.3) by age 16 years, and BMI-for-age z-scores ranged from 0 to 1 (IQR -0.7, 1.3) up to 8 years, and >1 (IQR -0.2, 1.9) until 16 years. The percentage of overweight and obesity was lowest in children aged < 5 years, and consistently > 40% in patients aged between 7 to 16 years. The prescribed total protein intake was associated with improved height growth (p < 0.01). Impaired growth in early life improved with age achieving normal population standards. Further studies are needed to investigate factors that influence growth outcome in HTI patients.

Characterization of Human Genes Modulated by Porphyromonas gingivalis Highlights the Ribosome, Hypothalamus, and Cholinergic Neurons.

Porphyromonas gingivalis, a bacterium associated with periodontal disease, is a suspected cause of Alzheimer's disease. This bacterium is reliant on gingipain proteases, which cleave host proteins after arginine and lysine residues. To characterize gingipain susceptibility, we performed enrichment analyses of arginine and lysine proportion proteome-wide. Genes differentially expressed in brain samples with detected P. gingivalis reads were also examined. Genes from these analyses were tested for functional enrichment and specific neuroanatomical expression patterns. Proteins in the SRP-dependent cotranslational protein targeting to membrane pathway were enriched for these residues and previously associated with periodontal and Alzheimer's disease. These ribosomal genes are up-regulated in prefrontal cortex samples with detected P. gingivalis sequences. Other differentially expressed genes have been previously associated with dementia (ITM2B, MAPT, ZNF267, and DHX37). For an anatomical perspective, we characterized the expression of the P. gingivalis associated genes in the mouse and human brain. This analysis highlighted the hypothalamus, cholinergic neurons, and the basal forebrain. Our results suggest markers of neural P. gingivalis infection and link the cholinergic and gingipain hypotheses of Alzheimer's disease.

Unacceptable Antibiotic Resistance Rates for Helicobacter pylori in Turkey: Something Must Change.

BACKGROUND: It is known that clarithromycin resistance has increased over the years (success rate 60%). The aim of the study was to investigate the importance of regional antimicrobial resistance rates for full accuracy of both diagnosis and treatment of Helicobacter pylori infection. METHODS: This study was carried out in the University Hospital Department of Gastroenterology. A total of 116 patients were evaluated with upper gastrointestinal endoscopy. Gastric antrum and corpus biopsy samples were taken for the rapid urease test (RUT), culture, and antimicrobial susceptibility testing for the presence of H. pylori. Antimicrobial susceptibilities of isolated H. pylori strains for clarithromycin and levofloxacin were determined by the epsilometer test (E-test). Minimal inhibitory concentration values for clarithromycin and levofloxacin were ≥1 and >1 µg/mL, respectively. RESULTS: H. pylori infection was considered clinically positive in 93 (80.2%) patients with either the RUT, culture, or histopathological examination. Seventy (60.3%) of the patients had RUT positivity. Sixty (85.7%) of these 70 patients had RUT positivity within the first 20 min. Among the 90 patients, who had a histopathological examination, HLO was positive in 76 (84.4%) patients. Fifty-two (44.8%) out of 116 patients were culture positive. Resistance rates for both clarithromycin and levofloxacin were high. In these 52 culture-positive patients, resistance rates determined for clarithromycin and levofloxacin were 26.9% and 25.5%, respectively. CONCLUSION: Clarithromycin or levofloxacin-based treatment regimen may not be an ideal alternative therapy for Turkish patients regardless of culture.

MicroRNAs as biomarkers for breast cancer.

Breast cancer is the most common type of cancer among women and the most frequent cause of death due to cancer among women. The lack of standard biomarkers in the early diagnosis of breast cancer, microRNAs (miRNA) have been of interest recently. Although, miRNAs are 19-24 nucleotide-long non-coding RNA species, they have crucial roles in many areas from organogenesis to carcinogenesis. This study has been conducted to investigate miR 21, miR 27b, miR 125a, miR 155, miR 200c, miR 335 miR373 as biomarkers in the early diagnosis of breast cancer; a selection based on the literature. Two miRNAs, miR 181 and miR 192 were selected as the endogenous control. MiRNAs were obtained from 5 cc blood samples taken from 20 breast cancer patients and 20 healthy people. 10 microRNAs were studied using Real Time PCR method. As a result, the quantities of miR 21, miR155 and miR125 ​​were significantly higher in the breast cancer group than in healthy controls. We suggest that performing validation studies in wider populations can help the use of miRNAs as biomarkers in the early diagnosis of breast cancer.