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Theoretically, a necrotic root canal fulfils all requirements as a niche for methanogens to inhabit. However, their presence in it and its implication in apical periodontitis (AP) is controversial. Therefore, to contribute to ending the controversy, this study aimed to detect and compare methanogens' presence in two distinct niches with supposedly different microenvironments; both were necrotic root canals associated with AP but one from patients with type 2 diabetes mellitus (T2DM) while the other from non-diabetic patients. A clinical examination was performed on 65 T2DM patients and 73 non-diabetic controls. Samples from necrotic root canals were obtained, and methanogens were identified. The presence of methanogens was three times higher (27.6%) in the T2DM group than in non-diabetic patients (8.2%). In addition, methanogens' presence was associated with a higher prevalence of periapical symptoms., (© 2023 The Authors. Australian Endodontic Journal published by John Wiley & Sons Australia, Ltd on behalf of Australian Society of Endodontology Inc.)

It has been proposed that oral commensal bacteria are potential reservoirs of a wide variety of antimicrobial resistance genes (ARGs) and could be the source of pathogenic bacteria; however, there is scarce information regarding this. In this study, three common streptococci of the mitis group ( S. oralis , S. sanguinis , and S. gordonii ) isolated from dental plaque (DP) were screened to identify if they were frequent reservoirs of specific ARGs ( blaTEM , cfxA , tetM , tetW , tetQ , ermA , ermB , and ermC ). DP samples were collected from 80 adults; one part of the sample was cultured, and from the other part DNA was obtained for first screening of the three streptococci species and the ARGs of interest. Selected samples were plated and colonies were selected for molecular identification. Thirty identified species were screened for the presence of the ARGs. From those selected, all of the S. sanguinis and S. oralis carried at least three, while only 30% of S. gordonii strains carried three or more. The most prevalent were tetM in 73%, and blaTEM and tetW both in 66.6%. On the other hand, ermA and cfxA were not present. Oral streptococci from the mitis group could be considered frequent reservoirs of specifically tetM , blaTEM , and tetW . In contrast, these three species appear not to be reservoirs of ermA and cfxA .

Background: Antimicrobial resistance is a global public health problem. Root canal microbiota associated with apical periodontitis represents a well-known reservoir of antimicrobial resistance genes (ARGs). However, the effect of type 2 diabetes mellitus (T2DM) in this reservoir is unknown. This study aimed to establish if root canal microbiota associated with apical periodontitis in T2DM patients is an augmented reservoir by identifying the prevalence of nine common ARGs and comparing it with the prevalence in nondiabetic patients., Methodology: This cross-sectional study included two groups: A T2DM group conformed of 20 patients with at least ten years of living with T2DM and a control group of 30 nondiabetic participants. Premolar or molar teeth with pulp necrosis and apical periodontitis were included. A sample was collected from each root canal before endodontic treatment. DNA was extracted, and ARGs were identified by polymerase chain reaction., Results: tetW and tetM genes were the most frequent (93.3 and 91.6%, respectively), while ermA was the least frequent (8.3%) in the total population. The distribution of the ARGs was similar in both groups, but a significant difference (p<0.005) was present in ermB, ermC, cfxA, and tetQ genes, being more frequent in the T2DM group. A total of eighty percent of the T2DM patients presented a minimum of four ARGs, while 76.6% of the control group presented a maximum of three., Conclusions: Root canal microbiota associated with apical periodontitis in T2DM patients carries more ARGs. Therefore, this pathological niche could be considered an augmented reservoir.

Cruciferous vegetables such as cauliflower and radish contain isothiocyanates exhibiting chemoprotective effects in vitro and in vivo. This research aimed to assess the impact of cauliflower (CIE) and radish (RIE) isothiocyanate extracts on the metabolic activity, intracellular reactive oxygen species (ROS), and LDH production of selected human colorectal adenocarcinoma cells (HCT116 and HT-29 for early and late colon cancer development, respectively). Non-cancerous colon cells (CCD-33Co) were used as a cytotoxicity control. The CIE samples displayed the highest allyl isothiocyanate (AITC: 12.55 µg/g) contents, whereas RIE was the most abundant in benzyl isothiocyanate (BITC: 15.35 µg/g). Both extracts effectively inhibited HCT116 and HT-29 metabolic activity, but the CIE impact was higher than that of RIE on HCT116 (IC 50 : 0.56 mg/mL). Assays using the half-inhibitory concentrations (IC 50 ) of all treatments, including AITC and BITC, displayed increased ( p < 0.05) LDH (absorbance: 0.25-0.40 nm) and ROS release (1190-1697 relative fluorescence units) in both cell lines. BITC showed the highest in silico binding affinity with all the tested colorectal cancer molecular markers (NF-kB, β-catenin, and NRF2-NFE2). The theoretical evaluation of AITC and BITC bioavailability showed high values for both compounds. The results indicate that CIE and RIE extracts display chemopreventive effects in vitro, but additional experiments are needed to validate their effects.

Vaccines against bovine babesiosis must, ideally, induce a humoral immune response characterized by neutralizing antibodies against conserved epitopes and a cellular Th1 immune response. In Babesia bovis, proteins such as AMA-1, MSA-2c, and RAP-1 have been characterized and antibodies against these proteins have shown a neutralizing effect, demonstrating the implication of B and T-cell epitopes in the immune response. There is evidence of the existence of B and T-cell epitopes in these proteins, however, it remains to be defined, the presence of conserved peptides in strains from around the world containing B and T-cell epitopes, and their role in the generation of a long-lasting immunity. The aim in this paper was to identify peptides of Babesia bovis AMA-1, MSA-2c, and RAP-1 that elicit a neutralizing and long-lasting Th1 immune response. Peptides containing B-cell epitopes of AMA-1, MSA-2c and RAP-1, were identified. The immune response generated by each peptide was characterized in cattle. All peptides tested induced antibodies that recognized intraerythrocytic parasites, however, only 5 peptides generated neutralizing antibodies in vitro: P2AMA-1 (6.28%), P3MSA-2c (10.27%), P4MSA-2c (10.42%), P1RAP-1 (32.45%), and P4RAP-1 (36.98%). When these neutralizing antibodies were evaluated as a pool, the inhibition percentage of invasion increased to 52.37%. When the T cellular response was evaluated, two peptides: P3MSA2c and P2AMA1 induced a higher percentage (>70%) of activated CD4 +/CD45RO+ T cells than unstimulated cells. Additionally, both peptides induced the production of gamma interferon (IFN-) in PBMCs from vaccinated cattle after one year proving the implication of a long-lasting Th1 immune response. In conclusion, we identified conserved peptides containing B and T-cell epitopes in antigens of B. bovis that elicit a Th1 immune response and showed evidence that peptides from the same protein elicit different immune responses, which has implication for vaccine development in bovine babesiosis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Endodontic freshly mixed sealers display toxic effects; however, these are significantly reduced and most become relatively inert in the set state but there is no information about the possible inflammatory reaction promoted by them. Four contemporary and different formulated endodontic set sealers (MTA Fillapex, BioRoot RCS, AH Plus, and Pulp Canal Sealer) were evaluated. Human periodontal ligament cells and human peripheral blood mononuclear cells were stimulated for 3, 6, 12 and 24 h. Interleukin-6, tumour necrosis factor-alpha, interleukin-8 and interleukin-10 concentrations were measured by enzyme-linked immunosorbent assay. All endodontic set sealer eluates promoted a similar production (P ˃ 0.05) of the four cytokines. However, their concentrations decreased within a short time period to nearly undetectable concentrations after 24 h, suggesting that the studied endodontic set sealers do not possess inflammatory properties which has favoured their long-term use in clinical practice., (© 2021 Australian Society of Endodontology Inc.)

Objective: This investigation aimed to detect coincidences in the antimicrobial resistance genes (ARG) profiles between members of a group living in a household and to compare them between other groups in order to establish if an exchange of ARG occurs and if dental plaque microbiota can be considered as a source and reservoir of ARG that can be shared between humans and pets., Methods: One hundred sixty dental plaque samples were obtained from four groups: Shelter dogs group (n=20), adult pet owners and dogs group (AD group, n=40), adult pet owners, children and dogs group (ACD group, n=60), and adult non-pet owners and children group (AC group, n=40). DNA was obtained, and specific primers with polymerase chain reaction for ARG detection were used., Results: The AD group exhibited the most coincidences in their ARG profiles, 14 (70%) of the 20 profiles coincided in 100% followed by the ACD group with 9 (45%) coincidences. While the AC group was the less coincident group, only 7 (35%) of the 20 profiles coincided. tetM was the most prevalent with 53.1%, followed by tetQ with 52.5% and cfxA with 51.2%, while the less prevalent were tetW with 31.8%, blaTEM-1 with 27.5%, and ermC with 18.7%., Conclusion: Dental plaque microbiota can be considered as a source and reservoir of ARG that can be shared between humans and dogs living in a household. The dogs seem to play an important role in the transference of ARG, and the children appear to be the most affected by carrying the most significant number of ARG., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

To identify the prevalence of C. albicans in primary endodontic infections of type two diabetes mellitus (T2DM) patients and compare their clinical and radiographical characteristics with a non-diabetic control group, establishing the possible relationship between primary endodontic infection, T2DM, and C. albicans, since diabetes mellitus (DM), influences the development, course, and response to the treatment of apical periodontitis, but the presence of Candida albicans (C. albicans) has not been considered before. A total of 120 patients were selected and divided into two groups: 60 T2DM diagnosed patients and 60 non-diabetic controls. A clinical examination and radiographic analysis were performed to establish a periapical index score (PAI). Root canal samples were taken. Deoxyribonucleic acid (DNA) was extracted, and specific primers were used to identify C. albicans by polymerase chain reaction (PCR). A twofold increase in the prevalence of C. albicans in T2DM patients was observed in contrast to control patients (p = 0.0251). Sixty-five percent of T2DM patients with positive C. albicans scored a ≥ 3 PAI, while only 27% of the patients without C. albicans had a ≥ 3 PAI score (p = 0.0065). Long-term DM patients presented C. albicans more frequently (p < 0.0001). In this study, long-term T2DM patients carried C. albicans in their root canals more frequently when having a primary endodontic infection. Furthermore, this C. albicans presence seems to be related to a higher frequency of apical periodontitis.

Background and Objectives : The knowledge that the cholinergic neurons from pedunculopontine nucleus (PPN) are vulnerable to the degeneration in early stages of the Parkinson disease progression has opened new perspectives to the development of experimental model focused in pontine lesions that could increase the risk of nigral degeneration. In this context it is known that PPN lesioned rats exhibit early changes in the gene expression of proteins responsible for dopaminergic homeostasis. At the same time, it is known that nicotinic cholinergic receptors (nAChRs) mediate the excitatory influence of pontine-nigral projection. However, the effect of PPN injury on the expression of transcription factors that modulate dopaminergic neurotransmission in the adult brain as well as the α7 nAChRs gene expression has not been studied. The main objective of the present work was the study of the effects of the unilateral neurotoxic lesion of PPN in nuclear receptor-related factor 1 (Nurr1), paired-like homeodomain transcription factor 3 (Pitx3), and α7 nAChRs mRNA expression in nigral tissue. Materials and Methods : The molecular biology studies were performed by means of RT-PCR. The following experimental groups were organized: Non-treated rats, N-methyl-D-aspartate (NMDA)-lesioned rats, and Sham operated rats. Experimental subjects were sacrificed 24 h, 48 h and seven days after PPN lesion. Results : Nurr1 mRNA expression, showed a significant increase both 24 h ( p < 0.001) and 48 h ( p < 0.01) after PPN injury. Pitx3 mRNA expression evidenced a significant increase 24 h ( p < 0.001) followed by a significant decrease 48 h and seven days after PPN lesion ( p < 0.01). Finally, the α7 nAChRs nigral mRNA expression remained significantly diminished 24 h, 48 h ( p < 0.001), and 7 days ( p < 0.01) after PPN neurotoxic injury. Conclusion : Taking together these modifications could represent early warning signals and could be the preamble to nigral neurodegeneration events., Competing Interests: The authors declare no conflict of interest.

Objectives: The aim of this study was to identify the prevalence of genes encoding resistance to three groups of antimicrobial agents in root canal samples from primary infection or post-treatment disease in Mexico., Methods: A total of 64 subjects requiring root canal treatment because of primary infection or post-treatment disease were enrolled in this cross-sectional analytical study. Root canal samples were obtained and DNA was isolated. Specific primers for six antimicrobial resistance genes (ARGs) and seven bacterial taxa (five genera and two species) were used. Student's t-test, χ 2 test and Fisher's exact test were applied where appropriate to detect statistical differences., Results: The bla TEM-1 , ermC and tetM genes were found more frequently in the post-treatment disease group compared with the primary infection group. The occurrence of assessed bacteria was similar in both groups, except for Enterococcus spp. and Porphyromonas endodontalis, which were found at a significantly higher frequency in the post-treatment disease group. It was observed that the post-treatment disease group harboured more ARGs. The most frequent ARG was tetW, whereas tetQ and cfxA were not detected in any case. With respect to bacterial taxa, Fusobacterium spp. was present in 100% of samples, whereas Porphyromonas gingivalis was not observed in any of the samples., Conclusions: At least one ARG was detected in all cases; moreover, 32.8% of samples were positive for four ARGs, 54.7% for three ARGs, 9.4% for two ARGs and only 3.1% for one ARG. This indicates a high prevalence and diversity of ARGs in these root canal samples., (Copyright © 2018 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

Purpose: The aim of this work was to evaluate the effect of PPAR agonists on the differentiation and metabolic features of porcine mesenchymal stem cells induced to the adipogenic or myogenic lineages., Methods: Bone marrow MSCs from neonate pigs were isolated and identified by cell proliferation, cell surface markers or the gene expression of stem cells (CD44, CD90, CD105 or Oct4 and Nanog, respectively). Cells were differentiated into adipose or muscle cells and treated with the PPAR agonists; adipogenic and myogenic differentiation was promoted by adding these compounds. The expression of PPARγ (an adipose marker) and MyoD1 and MyHC (muscle markers), metabolic changes and expression levels of metabolic enzymes involved in glycolysis, lipogenesis, lipolysis and the pentose phosphate pathway were tested by qPCR., Results: MSCs from neonate pigs exhibited high proliferation and were positive for CD44, CD90 and CD105 markers and Oct4 and Nanog expression. The treatment that promoted the highest expression of PPARγ was 50 µM of conjugated linoleic acid (CLA) c9 t11 (6.44 ± 0.69-fold, p ≤ 0.0001) in the adipose differentiation, and upregulation of HX2, ACCAα, ATGL, LPL and G6DP (p ≤ 0.0001) and downregulation of PFK and ACCAβ (p ≤ 0.0001) were found. For muscle differentiation, the best treatment was 50 µM of CLA c10 t12 (59.72 ± 4.72-fold, p ≤ 0.0001), and metabolic changes were upregulation of PFK, ACCAβ, G6DP, CPT1 and PPARβ/δ (p ≤ 0.0001), but no effect was observed with HX2 and ACCAα (p ≥ 0.05)., Conclusions: Our results suggest that differentiated cells exhibit a typical cell lineage metabolism and higher efficiencies both in anabolism and catabolism., (© 2016 S. Karger AG, Basel.)