This study evaluated the capacity of 23 multidrug-resistant (MDR) clinical isolates of Acinetobacter baumannii to adhere to respiratory epithelial cell surfaces and to form biofilm on a polystyrene ...surface. All 23 A. baumannii isolates were capable of adhering efficiently to respiratory epithelial cells, and biofilm production was positively associated with epithelial cell adhesiveness (r 0.80, p <0.0001). In the presence of the chelating agent EDTA, biofilm formation was markedly reduced. Cell adhesiveness and biofilm formation were significantly higher in isolates carrying the blaPER-1 gene as compared with isolates without this extended-spectrum β-lactamase gene (p <0.005 and p <0.001, respectively). Further examination by RT-PCR showed a positive correlation between the level of expression of the blaPER-1 gene and the level of biofilm formation (r 0.89, p <0.0001) and cell adhesiveness (r 0.74, p <0.006). Overall, the study demonstrated a high capacity of clinical isolates of MDR A. baumannii to form biofilm and to adhere to respiratory epithelial cells. This feature, combined with multidrug resistance, might contribute to the survival of these organisms and their dissemination in the hospital environment.
Aim
To compare the mineralization inductive capacity of Biodentine and Bioaggregate with Mineral trioxide aggregate (MTA) and to investigate possible signaling pathways of mineralization in human ...dental pulp cells (HDPCs).
Methodology
Viability of HDPCs in response to Biodentine, Bioaggregate, and MTA was measured using 3‐4,5‐dimethylthiazol‐2‐yl‐2,5 diphenyltetrazolium bromide. To investigate their potential to induce odontoblast differentiation, expression of dentine sialophosphoprotein (DSPP) and dentine matrix protein1 (DMP1) mRNA level was evaluated by RT‐PCR. For the mineralized nodule assay, Alizarin red staining was performed. To determine the role of MAPK signaling in the odontoblastic differentiation of HDPCs, activated MAPKs were investigated by Western blot and the effect of MAPK inhibitor was examined by Alizarin red S staining. The results were statistically analysed using one‐way anova and the Bonferroni test.
Results
The effects of MTA, Biodentine, and Bioaggregate on cell viability were similar. Biodentine and Bioaggregate enhanced DSPP and DMP1 mRNA expression compared to the control group, but to the same extent as MTA (P < 0.05). MTA, Biodentine, and Bioaggregate increased the area of calcified nodules compared to the control (P < 0.01). MTA, Biodentine, and Bioaggregate increased phosphorylation of extracellular signal‐regulated kinase (ERK), p38, and c‐Jun N‐terminal kinase (JNK). MAPK inhibitors attenuated mineralized nodule formation, which was increased by MTA, Biodentine, and Bioaggregate, respectively (P < 0.01).
Conclusion
Biodentine and Bioaggregate stimulated odontoblastic differentiation and mineralization nodule formation by activating the MAPK pathway as did MTA. This suggests that the new materials could be useful for regenerative endodontic procedures.
A set of mutated Encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES) elements with varying strengths is generated by mutating the translation initiation codons of 10(th), 11(th), ...and 12(th) AUG to non-AUG triplets. They are able to control the relative expression of multiple genes over a wide range in mammalian cells in both transient and stable transfections. The relative strength of each IRES mutant remains similar in different mammalian cell lines and is not gene specific. The expressed proteins have correct molecular weights. Optimization of light chain over heavy chain expression by these IRES mutants enhances monoclonal antibody expression level and quality in stable transfections. Uses of this set of IRES mutants can be extended to other applications such as synthetic biology, investigating interactions between proteins and its complexes, cell engineering, multi-subunit protein production, gene therapy, and reprogramming of somatic cells into stem cells.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Epitheliotropic intestinal T-cell lymphoma (EITL, also known as type II enteropathy-associated T-cell lymphoma) is an aggressive intestinal disease with poor prognosis and its molecular alterations ...have not been comprehensively characterized. We aimed to identify actionable easy-to-screen alterations that would allow better diagnostics and/or treatment of this deadly disease. By performing whole-exome sequencing of four EITL tumor-normal pairs, followed by amplicon deep sequencing of 42 tumor samples, frequent alterations of the JAK-STAT and G-protein-coupled receptor (GPCR) signaling pathways were discovered in a large portion of samples. Specifically, STAT5B was mutated in a remarkable 63% of cases, JAK3 in 35% and GNAI2 in 24%, with the majority occurring at known activating hotspots in key functional domains. Moreover, STAT5B locus carried copy-neutral loss of heterozygosity resulting in the duplication of the mutant copy, suggesting the importance of mutant STAT5B dosage for the development of EITL. Dysregulation of the JAK-STAT and GPCR pathways was also supported by gene expression profiling and further verified in patient tumor samples. In vitro overexpression of GNAI2 mutants led to the upregulation of pERK1/2, a member of MEK-ERK pathway. Notably, inhibitors of both JAK-STAT and MEK-ERK pathways effectively reduced viability of patient-derived primary EITL cells, indicating potential therapeutic strategies for this neoplasm with no effective treatment currently available.
•An experimental study of thermochemical conversion of microalgae was carried out.•Energy balance and efficiency of co-products char, bio-oil and gas was determined.•Energy balance equations were ...established for process life cycle efficiencies.•Comparisons were made with other energy indicators from literature.•Life cycle CO2 results displayed more CO2 emissions released than absorbed.
An investigation of the potential to efficiently convert lipid-depleted residual microalgae biomass using thermochemical (gasification at 850°C, pyrolysis at 550°C, and torrefaction at 300°C) processes to produce bioenergy derivatives was made. Energy indicators are established to account for the amount of energy inputs that have to be supplied to the system in order to gain 1MJ of bio-energy output. The paper seeks to address the difference between net energy input–output balances based on a life cycle approach, from “cradle-to-bioenergy co-products”, vs. thermochemical processes alone. The experimental results showed the lowest results of Net Energy Balances (NEB) to be 0.57MJ/MJ bio-oil via pyrolysis, and highest, 6.48MJ/MJ for gas derived via torrefaction. With the complete life cycle process chain factored in, the energy balances of NEBLCA increased to 1.67MJ/MJ (bio-oil) and 7.01MJ/MJ (gas). Energy efficiencies and the life cycle CO2 emissions were also calculated.
Meta-analyses of genome-wide association studies (GWAS) have identified more than 240 loci that are associated with type 2 diabetes (T2D)
; however, most of these loci have been identified in ...analyses of individuals with European ancestry. Here, to examine T2D risk in East Asian individuals, we carried out a meta-analysis of GWAS data from 77,418 individuals with T2D and 356,122 healthy control individuals. In the main analysis, we identified 301 distinct association signals at 183 loci, and across T2D association models with and without consideration of body mass index and sex, we identified 61 loci that are newly implicated in predisposition to T2D. Common variants associated with T2D in both East Asian and European populations exhibited strongly correlated effect sizes. Previously undescribed associations include signals in or near GDAP1, PTF1A, SIX3, ALDH2, a microRNA cluster, and genes that affect the differentiation of muscle and adipose cells
. At another locus, expression quantitative trait loci at two overlapping T2D signals affect two genes-NKX6-3 and ANK1-in different tissues
. Association studies in diverse populations identify additional loci and elucidate disease-associated genes, biology, and pathways.
Summary
Understanding the natural history of lateral femoral stress fractures helps to guide their management. Improvement in their radiographic characteristics is rare. Progression was generally ...sequential, most developing an incomplete fracture line before fracture displacement. Stopping bisphosphonates decreased the fracture rate, a feasible management option for lesions without incomplete fracture lines.
Introduction
Retrospective study evaluating the natural history of lateral femoral stress fractures (FSF) by serial radiography over a variable period of time in a cohort of patients treated for some time with bisphosphonates for osteoporosis, whilst also identifying the fracture response in cases where bisphosphonates were discontinued.
Methods
The radiographs of 76 consecutive patients (92 femurs) with 161 FSF were reviewed to document their change over time. Femurs were classified into the following: A—normal, B—focal cortical thickening, C—dreaded black line and D—displaced fracture. Bisphosphonate history was recorded.
Results
66.5% FSF showed group stability between the first and last radiographs: group B (79.1%), group C (45.7%). 28.6% progressed, mostly following an ordered sequence starting from group A, progressing to B, then C, before culminating in D. Progression rate was as follows: A—100% (11/11), B—18.3% (21/115), C—40% (14/35). Regression in FSF was uncommon—5.6% (8/161). 34.8% (32/92) sustained displaced fractures. Kaplan-Meier analysis showed statistically significant difference between the groups; median survival (95% CI): A—4189 (-), B—3383.0 (-), C—1807 (0.0–3788.6) and progression to displaced fracture when bisphosphonate had been stopped for at least 6 months. The group without recent bisphosphonates had a lower group progression rate (17.1%, 12/70). Nevertheless, 10.9% (5/46) progressed to displaced fracture. This group also had the highest proportion of stable (77.1%, 54/70) and regressive lesions (5.7%, 4/70).
Conclusions
In FSF, there is natural progression from normal bone, to focal cortical thickening, to dreaded black line and eventually to displaced fracture. Most lesions persist, remaining static or progressing, especially if a dreaded black line is present and bisphosphonates are continued. Regression is uncommon and more frequent when bisphosphonates are discontinued. Despite stopping bisphosphonates, there remains a 10.9% risk of progression to displaced fracture.
Despite the well-recognized importance of caries risk assessment, practical models remain to be established. This study was designed to develop biopsychosocial models for caries risk assessment in ...various settings. With a questionnaire, an oral examination, and biological (salivary, microbiological, and plaque pH) tests, a prospective study was conducted among 1782 children aged 3-6 years, with 1576 (88.4%) participants followed in 12 months. Multiple risk factors, indicators, and protective factors were identified. Various risk assessment models were constructed by the random selection of 50% of the cases and further validated in the remaining cases. For the prediction of a “one-year caries increment”, screening models without biological tests achieved a sensitivity/specificity of 82%/73%; with biological tests, full-blown models achieved the sensitivity/specificity of 90%/90%. For identification of a quarter of the children with high caries burden (baseline dmft > 2), a community-screening model requiring only a questionnaire reached a sensitivity/specificity of 82%/81%. These models are promising tools for cost-effective caries control and evidence-based treatment planning. Abbreviations: decayed, missing, filled teeth in primary dentition (dmft); receiver operation characteristics (ROC); relative risk (RR); confidence interval (CI); National Institutes of Health (NIH); World Health Organization (WHO); US Department of Health and Human Services (US/DHHS); American Academy of Pediatric Dentistry (AAPD).
This paper presents a modelling method that seeks to support the prediction and management of undesired engineering change (EC) propagation during the design and development of complex products. The ...method builds on the house of quality and the change prediction method to model the effects of potential change propagation brought about by different change options. The objective is to better reflect how well each change option can address the product requirements. The method was applied to the design of a jet engine fan during a case study with an aerospace company. The findings suggest that this modelling approach is suitable for assessing the effects of potential EC propagation and can support companies in effective exploration of the design space.
Aim
To explore the involvement of TLR5 in pulp inflammation and to examine the effects of TLR5 activation with its ligand, FlaB protein, on pro‐inflammatory gene expression.
Methodology
TLR5 ...expression in dental pulp tissues and human dental pulp cells (hDPCs) were determined by immunohistochemistry, immunocytochemistry, Western blots and RT‐PCR analyses. To examine the role of TLR5, hDPCs were treated with recombinant FlaB protein (500 ng mL−1) to activate the receptor or with a small interfering RNA against TLR5 (si‐TLR5) to downregulate the receptor. After exposure to FlaB, the expression of inflammation‐related proteins was screened using a protein array kit. Western blots or qRT‐PCR analyses were performed to identify changes in the expression of uPA (urokinase plasminogen activator), TIMPs (tissue inhibitor of metalloproteinases), and IL‐6 and to determine their signalling pathways. Statistical analysis was performed using one‐way analysis of variance (anova) with Tukey post hoc test; P < 0.05 was considered statistically significant.
Result
TLR5 expression was identified in pulp tissues and hDPCs. In the protein array analysis, treatment with FlaB significantly increased uPA expression (P < 0.01) and significantly decreased TIMP1/4 (P < 0.05). FlaB treatment also significantly increased expression of the inflammatory marker IL‐6 (P < 0.01). FlaB treatment increased phosphorylation of the NF‐κB p65 subunit, JNK, p38 and ERK. Chemical inhibitors of NF‐κB (Bay11‐7082), p38 (SB202190) or ERK (U0126) decreased the FlaB induction of uPA expression. Downregulation of TLR5 expression by siRNA decreased the FlaB induction of uPA protein and p65 phosphorylation.
Conclusion
TLR5 activation with FlaB treatment induced the expression of uPA via the NF‐κB and MAPK signalling pathways. Flagellin‐bearing oral bacteria may cause pulp inflammation through TLR5. The findings provide new clues to control pulpal diseases by targeting TLR5 signalling pathways.
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