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Background: Many studies have focused on the significance of lipid regulatory genes in the pathophysiology of Coronary artery disease (CAD). ApoB XbaI (rs693) and EcoRI (rs1042031) single nucleoid polymorphisms (SNPs) were investigated to detect whether they are risk factors for CAD. Till now, this association remains uncertain. SMARCA4 (rs1122608) SNP has directly related to dyslipidemia. Loss of function mutations (LOF) in PCSK9 result in a reduction in LDL cholesterol and are associated with protection from the development of CAD., Methods: This study was conducted on 54 CAD patients who were admitted at Internal Medicine Specialized Hospital (Cardiology Department) and 47 healthy controls. Peripheral blood samples were taken from both groups. DNA was extracted from EDTA-blood samples, then PCR- RFLP for ApoB XbaI (rs693) and EcoRI (rs1042031), SMARCA4 (rs1122608) and PCSK9 (rs505151) SNPs was done., Results: No statistically significant difference was found between patients and controls as regard EcoRI SNP. XbaI (rs693) X + X + genotype was significantly higher in control group (P = 0.0355). SMARCA4 (TT, GT + TT) genotypes, and T allele (P < 0.001); PCSK9 AG genotype and G allele (P = 0.027 and 0.032 respectively) were more frequent in CAD patients than controls., Conclusion: SMARCA4 (rs1122608) and PCSK9 (rs505151) SNPs are significantly accompanying with the risk of CAD development in the Egyptian population. X + X + genotype appeared to have a protective effect against CAD. However, no observed association between EcoRI (rs1042031) and the risk of CAD development was found., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Chitosan-based nanocomposites (CS NCs) are gaining considerable attention as multifaceted antifungal agents. This study investigated the antifungal activity of NCs against two phytopathogenic strains: Fusarium solani (F. solani) and Alternaria solani (A. solani). Moreover, it sheds light on their underlying mechanisms of action. The NCs, CS-ZnO, CS-CuO, and CS-SiO 2 , were characterized using advanced methods. Dynamic and electrophoretic light scattering techniques revealed their size range (60-170 nm) and cationic nature, as indicated by the positive zeta potential values (from +16 to +22 mV). Transmission electron microscopy revealed the morphology of the NCs as agglomerates formed between the chitosan and oxide components. X-ray diffraction patterns confirmed crystalline structures with specific peaks indicating their constituents. Antifungal assessments using the agar diffusion technique demonstrated significant inhibitory effects of the NCs on both fungal strains (1.5 to 4-fold), surpassing the performance of the positive control, nystatin. Notably, the NCs exhibited superior antifungal potency, with CS-ZnO NCs being the most effective. A. solani was the most sensitive strain to the studied agents. Furthermore, the tested NCs induced oxidative stress in fungal cells, which elevated stress biomarker levels, such as superoxide dismutase (SOD) activity and protein carbonyl content (PCC), 2.5 and 6-fold for the most active CS-CuO in F. solani respectively. Additionally, they triggered membrane lipid peroxidation up to 3-fold higher compared to control, a process that potentially compromises membrane integrity. Laurdan fluorescence spectroscopy highlighted alterations in the molecular organization of fungal cell membranes induced by the NCs. CS-CuO NCs induced a membrane rigidifying effect, while CS-SiO 2 and CS-ZnO could rigidify membranes in A. solani and fluidize them in F. solani. In summary, this study provides an in-depth understanding of the interactions of CS-based NCs with two fungal strains, showing their antifungal activity and offering insights into their mechanisms of action. These findings emphasize the potential of these NCs as effective and versatile antifungal agents., 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 © 2024 Elsevier B.V. All rights reserved.)

Background: Coronary artery disease (CAD) is the main cause of death in Egypt. Many LDL-R gene locus single nucleotide polymorphisms (SNP) are found to be associated with the risk of CAD. This research aimed to assess the allelic and genotypic frequencies of rs1122608 SNP and their association with the extent of vessel affection and lipid profile in a population of Egyptians.100 CAD patients and 100 healthy controls of Egyptians were included. PCR-RFLP was used to genotype rs1122608 SNPs., Results: Significantly higher proportion of 'T' allele among patient (risk allele). This association is of low strength (ϕ lies between 0.1 and 0.3). A participant with 'T' allele has 1.95 times higher odds to exhibit CAD versus a participant with 'G' allele. Significantly higher proportion of 'T/T' genotype among cases versus control (risk genotype). This association is of low strength (Cramer's V lies between 0.1 and 0.3). A participant with 'T/T' genotype has 4.5 times higher odds to exhibit CAD versus a participant with 'G/G'. Gensini score showed no significant association with rs1122608 genotypes (p = 0.863)., Conclusions: The mutant GT and TT genotypes and minor T allele of rs1122608 were positively correlated with CAD and considered as independent risk factors for CAD., (© 2023. The Author(s).)

The present study explores for the first time the effect of hyperbaric oxygen (HBO) on gingival mesenchymal stem cells' (G-MSCs) gene expression profile, intracellular pathway activation, pluripotency, and differentiation potential under an experimental inflammatory setup. G-MSCs were isolated from five healthy individuals ( n = 5) and characterized. Single (24 h) or double (72 h) HBO stimulation (100% O2, 3 bar, 90 min) was performed under experimental inflammatory [IL-1β (1 ng/mL)/TNF-α (10 ng/mL)/IFN-γ (100 ng/mL)] and non-inflammatory micro-environment. Next Generation Sequencing and KEGG pathway enrichment analysis, G-MSCs' pluripotency gene expression, Wnt-/β-catenin pathway activation, proliferation, colony formation, and differentiation were investigated. G-MSCs demonstrated all mesenchymal stem/progenitor cells' characteristics. The beneficial effect of a single HBO stimulation was evident, with anti-inflammatory effects and induction of differentiation ( TLL1 , ID3 , BHLHE40 ), proliferation/cell survival ( BMF , ID3 , TXNIP , PDK4 , ABL2 ), migration ( ABL2 ) and osteogenic differentiation ( p < 0.05). A second HBO stimulation at 72 h had a detrimental effect, significantly increasing the inflammation-induced cellular stress and ROS accumulation through HMOX1 , BHLHE40 , and ARL4C amplification and pathway enrichment ( p < 0.05). Results outline a positive short-term single HBO anti-inflammatory, regenerative, and differentiation stimulatory effect on G-MSCs. A second (72 h) stimulation is detrimental to the same properties. The current results could open new perspectives in the clinical application of short-termed HBO induction in G-MSCs-mediated periodontal reparative/regenerative mechanisms.

Background: The regular use of gold nanoparticles (Au-NPs) may increase the likelihood of human exposure to these nanoparticles (NPs) and raises concerns about toxicity., Aim: This study investigated the short-term impact of exposure to Au-NPs on inducing cerebellar pathology in rats, and whether the dose or duration of exposure was more important., Methodology: The study used two concentrations of Au-NPs (25 and 50 particles per million) and 18 rats were randomly assigned to three groups. Assessments of the animals were done via behavioral, gene expression, histological, and immunohistochemistry analyses., Results: Both concentrations of Au-NPs caused cerebellar pathology, as assessed through the investigation test battery. The Au-NPs50 group displayed more injury and decreased mobility compared with the control and the Au-NPs25 group. The Au-NPs25 group showed an increase in supported rearing and significant up-regulation of the Rgc32 gene compared with the control. The Trkb gene was insignificantly up-regulated in both Au-NPs groups compared with the control., Conclusion: The study indicates that exposure to Au-NPs can cause cerebellar pathology in rats and that the toxicity is more dependent on dose than the duration of exposure. These findings have significant implications for the safe use of Au-NPs in various applications., (© 2023 The Author(s).)

Objectives: Sonic/ultrasonic devices are essential tools in today's endodontics. This prospective trial evaluated for the first time the impact of practitioners' proficiency levels and patient-related factors on complications associated with a high frequency polyamide sonic irrigant activation device., Methods: In total 334 patients (females:158, males:176; age:18-95 years) received in the course of their endodontic therapy an intracanal irrigation, using a high frequency polyamide sonic irrigant activation device, by practitioners of different proficiency levels (undergraduate students, general practitioners or endodontists). Intracanal bleeding (yes/no), postoperative pain (0-10 scale), emphysema (yes/no) and polyamide tip fractures (yes/no) were recorded and related to proficiency levels, age, gender, tooth type, smoking-status, systemic conditions affecting healing ability, baseline pain, swelling, fistula, sensitivity to percussion and diagnosis., Results: Intracanal bleeding was associated with patients' age (p<0.05), baseline pain level (OR = 1.14, 95%CI = 0.91-1.22) and baseline swelling (OR = 2.73, 95%CI = 0.14-0.99; p<0.05) but not proficiency level, gender, tooth type, smoking, systemic conditions, baseline fistula or sensitivity to percussion (p>0.05). Postoperative pain development was related to proficiency level (p<0.05) and baseline pain level (p<0.001), with no influence of age, gender, tooth type, smoking, systemic conditions, baseline fistula, swelling or sensitivity to percussion (p>0.05). Emphysema and polyamide tip fractures were not reported., Conclusions: Within the current study's limitations, younger patients with higher baseline pain and swelling, were associated with higher intracanal bleeding. Apart from higher postoperative pain observed with less experienced practitioners, proficiency level had no influence on bleeding, polyamide tip fracture or emphysema, endorsing the high frequency polyamide sonic irrigation device as a safe therapeutic device., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Hahn et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The prevalence of obesity has risen in the last decades, and it has caused massive health burdens on people's health, especially metabolic and cardiovascular issues. The risk of vitamin D insufficiency is increased by obesity, because adipose tissue alters both the requirements for and bioavailability of vitamin D. Exercise training is acknowledged as having a significant and long-term influence on body weight control; the favorable impact of exercise on obesity and obesity-related co-morbidities has been demonstrated via various mechanisms. The current work illustrated the effects of vitamin D supplementation and exercise on obesity induced by a high-fat diet (HFD) and hepatic steatosis in rats and explored how fatty acid transport protein-4 (FATP4) and Toll-like receptor-4 antibodies (TLR4) might be contributing factors to obesity and related hepatic steatosis. Thirty male albino rats were divided into five groups: group 1 was fed a normal-fat diet, group 2 was fed an HFD, group 3 was fed an HFD and given vitamin D supplementation, group 4 was fed an HFD and kept on exercise, and group 5 was fed an HFD, given vitamin D, and kept on exercise. The serum lipid profile adipokines, interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) were analyzed, and the pathological changes in adipose and liver tissues were examined. In addition, the messenger-ribonucleic acid (mRNA) expression of FATP4 and immunohistochemical expression of TLR4 in adipose and liver tissues were evaluated. Vitamin D supplementation and exercise improved HFD-induced weight gain and attenuated hepatic steatosis, along with improving the serum lipid profile, degree of inflammation, and serum adipokine levels. The expression of FATP4 and TLR4 in both adipose tissue and the liver was downregulated; it was noteworthy that the group that received vitamin D and was kept on exercise showed also improvement in the histopathological picture of this group. According to the findings of this research, the protective effect of vitamin D and exercise against obesity and HFD-induced hepatic steatosis is associated with the downregulation of FATP4 and TLR4, as well as a reduction in inflammation.

Objective: The present randomized controlled trial compares for the first time the vestibular socket therapy (VST) to the contour augmentation technique in the management of compromised fresh extraction sockets in the maxillary esthetic zone, regarding mid-facial soft tissue changes (primary outcome), mesial and distal papillae dimensions, horizontal soft tissue changes and labial bone plate thickness at apical, middle and coronal levels (secondary outcomes) over 1-year., Materials and Methods: Forty participants with single nonrestorable maxillary teeth in the esthetic zone were randomized into two groups; VST (test; n = 20) utilizing vestibular access for guided bone regeneration (GBR) with immediate implant placement, or contour augmentation (control; n = 20) undergoing an initial healing period followed by implant placement with GBR through a conventional access flap., Results: All implants were successfully osseo-integrated, except for one implant in the test group. VST showed significantly less mid-facial soft tissue changes of -0.53 ± 1.17 mm versus -1.87 ± 0.69 mm in the control group (p < 0.001). Similarly, changes in mesial papilla (test = -0.64 ± 0.95 mm, control = -1.20 ± 0.81 mm), distal papilla (test = -0.56 ± 1.17 mm, control = -1.26 ± 0.63 mm), horizontal soft-tissue (test = -0.82 ± 0.95 mm, control = -1.84 ± 0.88 mm; p < 0.05) were significantly less in VST. Intra-group comparisons demonstrated a significant increase in labial bone thickness, with no differences between groups. Regression analysis revealed a significant correlation between VST as well as increased coronal bone thickness with the reduction in mid-facial soft-tissue changes., Conclusion: The VST showed less soft-tissue changes and could represent an innovative technique for implant placement in the maxillary esthetic zone. Both techniques showed a high implant survival rate and increased bone thickness after 12 months., (© 2022 Wiley Periodicals LLC.)

Zinc oxide (ZnO) nanomaterials offer some promising antibacterial effects. In this study, a new form of ZnO is synthesized, named ZnO nanocluster bars (NCs). Herein, ZnO NCs, ZnO nanoparticles (NPs), ZnO coated with silica (ZnO-SiOA, ZnO-SiOB), and SiO2 NPs were prepared, characterized, and their antimicrobial and prooxidant activity were tested. The prooxidant activity of all nanomaterials was studied according to free-radical oxidation reactions (pH 7.4 and pH 8.5) in chemiluminescent model systems. Each form of new synthesized ZnO nanomaterials exhibited a unique behavior that varied from mild to strong prooxidant properties in the Fenton`s system. ZnO NPs and ZnO NCs showed strong antibacterial effects, ZnO-SiOA NPs did not show any antibacterial activity representing biocompatibility. All tested NMs also underwent oxidation by H2O2. ZnO NCs and ZnO NPs exhibited strong oxidation at pH 8.5 in the O2-. generating system. While, SiO2, ZnO-SiOA andZnO-SiOB possessed pronounced 60-80% antioxidant effects, SiO2 NPs acted as a definitive prooxidant which was not observed in other tests. ZnO NCs are strong oxidized, assuming that ZnO NCs provide a slower release of ZnO, which leads to having a stronger effect on bacterial strains.  Thus, ZnO NCs are an important antibacterial agent that could be an emergent replacement of traditional antibiotics.

Vitamins and omega-3 fatty acids (Ω3FA) modulate periodontitis-associated inflammatory processes. The aim of the current investigation was to evaluate associations of oral nutrient intake and corresponding serum metabolites with clinical severity of human periodontitis. Within the Food Chain Plus cohort, 373 periodontitis patients—245 without (POL) and 128 with tooth loss (PWL)—were matched to 373 controls based on sex, smoking habit, age and body mass index in a nested case-control design. The amount of oral intake of vitamins and Ω3FAs was assessed from nutritional data using a Food Frequency Questionnaire. Oral intake and circulatory bioavailability of vitamins and Ω3FA serum metabolomics were compared, using ultra-high-resolution mass spectrometry. Periodontitis patients exhibited a significantly higher oral intake of vitamin C and Ω3FA Docosapentaenoic acid (p < 0.05) compared to controls. Nutritional intake of vitamin C was higher in PWL, while the intake of Docosapentaenoic acid was increased in POL (p < 0.05) compared to controls. In accordance, serum levels of Docosapentaenoic acid were also increased in POL (p < 0.01) compared to controls. Vitamin C and the Ω3FA Docosapentaenoic acid might play a role in the pathophysiology of human periodontitis. Further studies on individualized nutritional intake and periodontitis progression and therapy are necessary.

Products containing Silver nanoparticles (Ag NPs) are becoming vastly used in our daily life. The widespread increased introduction of Ag NPs in many aspects of life has raised researchers' concerns regarding their safety and toxicity for biological and environmental life in the past few years. The current study aimed to explore the subsequent effects of Ag NPs withdrawal, following short-term oral administration. Eighteen rats were assigned randomly into three groups (control group "1" and AG NPs treated groups "2" and "3"; 6 animals each). The control group received normal food and tap water while groups 2 & 3 received 0.5 ml of a solution containing 25 ppm Ag NPs for 14 days. Group 2 rats were sacrificed on day 14 whereas group 3 was left for another 14 days of particle cessation followed by euthanasia on day 28. Functional assessment was done by liver enzyme assays, hydrogen peroxide activity, hepatic Bdnf expression, and P53 immunoreactivity. Hepatic tissue structural assessment was done via hematoxylin and eosin, periodic acid-Schiff as well as Masson's trichrome stains. The results revealed a significant elevation of Hydrogen peroxide in group 2 only compared to the control group. Hepatic Bdnf and liver enzymes were both insignificantly affected. Structural abnormalities and enhanced apoptosis in hepatic tissue were found 14 days after ceasing the nanoparticles. In conclusion: Structural and functional insults following Ag NPs oral administration continues after particle withdrawal, and interestingly they do not necessitate apparent reflection on liver enzyme assays., Competing Interests: 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., (© 2022 The Author(s).)

Peroxisome proliferator-activated receptor gamma (PPAR-γ) is ubiquitously expressed in testicular tissue and plays a crucial role in regulating various physiological processes. Pioglitazone (PIO) is one of the PPAR-γ agonists, having anti-oxidant and anti-inflammatory effects. Patients on gentamycin treatment may undergo serious side effects such as testicular damage. To the best of our knowledge, this was the first study to investigate the possible protective anti-inflammatory and anti-apoptotic effects of PIO on gentamycin-induced testicular damage. Fifty adult male Wistar albino rats included in the study as the control group (CTL) received normal saline; a gentamycin-induced testicular damage group (GM) received gentamycin (100 mg/kg); PIO5, PIO10, PIO20 groups received PIO at a dose of 5, 10, and 20 mg/ kg, respectively, for 21 days, and gentamycin was started at day 15 of the experiment for 6 days. The parameters of spermatozoa and histopathological alterations in the testes were significantly improved in the PIO20 group. Moreover, MDA levels, inflammatory mediators, and apoptotic Bax expression were decreased. The activity of glutathione peroxidase, catalase, total antioxidant capacity, and anti-apoptotic Bcl-2 genes expression were increased. It was concluded that PIO20 could protect against gentamycin-induced testicular damage in Wistar rats through its anti-oxidant, anti-inflammatory, and antiapoptotic effects.

Objective: The current study aimed to evaluate the effect of electronic cigarette (EC) aerosol, Cannabis, and conventional cigarettes smoke on gingival fibroblast/gingival mesenchymal stem cells' (GF/G-MSCs) of never smokers., Material and Methods: Human GF/G-MSCs (n = 32) were isolated and characterized using light microscopy, flow cytometry, and multilineage differentiation ability. Following the application of aerosol/smoke extracts, GF/G-MSCs were evaluated for cellular proliferation; colony-forming units (CFU-F) ability; cellular viability (using the MTT assay); mitochondrial depolarization using JC-1 dye; and genes' expression of ATM, p21, Oct4, and Nanog., Results: Colony-forming units and viability (OD 450 nm) were significantly reduced upon exposure to Cannabis (mean ± SD; 5.5 ± 1.5; p < .00001, 0.47 ± 0.21; p < .05) and cigarettes smoke (2.3 ± 1.2 p < .00001, 0.59 ± 0.13, p < .05), while EC aerosol showed no significant reduction (10.8 ± 2.5; p = .05, 1.27 ± 0.47; p > .05) compared to the control group (14.3 ± 3, 1.33 ± 0.12). Significantly upregulated expression of ATM, Oct4, and Nanog (gene copies/GADPH) was noticed with Cannabis (1.5 ± 0.42, 0.82 ± 0.44, and 1.54 ± 0.52, respectively) and cigarettes smoke (1.52 ± 0.75, 0.7 ± 0.14, and 1.48 ± 0.79, respectively; p < .05), whereas EC aerosol caused no statistically significant upregulation of these genes compared to the control group (0.63 ± 0.1, 0.31 ± 0.12, and 0.64 ± 0.46, respectively; p > .05). The p21 gene was not significantly downregulated in EC aerosol (1.22 ± 0.46), Cannabis (0.71 ± 0.24), and cigarettes smokes (0.83 ± 0.54) compared to the control group (p = .053, analysis of variance)., Conclusion: Cannabis and cigarettes smoke induce DNA damage and cellular dedifferentiation and negatively affect the cellular proliferation and viability of GF/G-MSCs of never smokers, whereas EC aerosol showed a significantly lower impact on these properties., (© 2021 The Authors. Journal of Periodontal Research published by John Wiley & Sons Ltd.)

Aim: To assess platelet-rich fibrin (PRF) with ascorbic acid (AA) versus PRF in intra-osseous defects of stage-III periodontitis patients., Methodology: Twenty stage-III/grade C periodontitis patients, with ≥ 3 mm intra-osseous defects, were randomized into test (open flap debridement (OFD)+AA/PRF; n = 10) and control (OFD+PRF; n = 10). Clinical attachment level (CAL; primary outcome), probing pocket depth (PPD), gingival recession depth (RD), full-mouth bleeding scores (FMBS), full-mouth plaque scores (FMPS), radiographic linear defect depth (RLDD) and radiographic defect bone density (RDBD) (secondary-outcomes) were examined at baseline, 3 and 6 months post-surgically., Results: OFD+AA/PRF and OFD+PRF demonstrated significant intragroup CAL gain and PPD reduction at 3 and 6 months (p < 0.001). OFD+AA/PRF and OFD+PRF showed no differences regarding FMBS or FMPS (p > 0.05). OFD+AA/PRF demonstrated significant RD reduction of 0.90 ± 0.50 mm and 0.80 ± 0.71 mm at 3 and 6 months, while OFD+PRF showed RD reduction of 0.10 ± 0.77 mm at 3 months, with an RD-increase of 0.20 ± 0.82 mm at 6 months (p < 0.05). OFD+AA/PRF and OFD+PRF demonstrated significant RLDD reduction (2.29 ± 0.61 mm and 1.63 ± 0.46 mm; p < 0.05) and RDBD-increase (14.61 ± 5.39% and 12.58 ± 5.03%; p > 0.05). Stepwise linear regression analysis showed that baseline RLDD and FMBS at 6 months were significant predictors of CAL reduction (p < 0.001)., Conclusions: OFD+PRF with/without AA significantly improved periodontal parameters 6 months post-surgically. Augmenting PRF with AA additionally enhanced gingival tissue gain and radiographic defect fill., Clinical Relevance: PRF, with or without AA, could significantly improve periodontal parameters. Supplementing PRF with AA could additionally augment radiographic linear defect fill and reduce gingival recession depth., (© 2021. The Author(s).)

This study investigates the impacts of zinc oxide nanoparticles: bare (ZnO NPs) and ZnO NPs coated with silicon shell (ZnO-Si NPs), on Pisum sativum L. under physiological and salt stress conditions. The experimental results revealed that the foliar spray with ZnO-Si NPs and 200 mg/L ZnO NPs did not influence the stomata structure, the membrane integrity, and the functions of both photosystems under physiological conditions, while 400 mg/L ZnO-Si NPs had beneficial effects on the effective quantum yield of photosystem II (PSII) and the photochemistry of photosystem I (PSI). On the contrary, small phytotoxic effects were registered after spraying with 400 mg/L ZnO NPs accompanied by stimulation of the cyclic electron flow around PSI and an increase of the non-photochemical quenching (NPQ). The results also showed that both types of NPs (with exception of 400 mg/L ZnO NPs) decrease the negative effects of 100 mM NaCl on the photochemistry of PSI (P700 photooxidation) and PSII (qp, Fv/Fm, Fv/Fo, Φ PSII, Φ exc ), as well as on the pigment content, stomata closure and membrane integrity. The protective effect was stronger after spraying with ZnO-Si NPs in comparison to ZnO NPs, which could be due to the presence of Si coating shell. The role of Si shell is discussed., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

Objective: The objective of this randomized controlled trial was to evaluate the radiographic changes and histologic healing following alveolar ridge preservation (ARP) using autogenous whole tooth (AWTG), test group, versus autogenous demineralized dentin graft (ADDG), control group., Material and Methods: Twenty non-molar teeth indicated for extraction were randomized into two groups (n = 10/group). Extracted teeth were prepared into AWTG or ADDG (0.6N HCl; 30 min), inserted into extraction sockets and covered by collagen membranes. Cone-beam computed tomography (CBCT) scans at baseline and six months were compared to assess ridge-dimensional changes. At six months, bone biopsies of engrafted sites were harvested and analyzed histomorphometrically., Results: All sites healed uneventfully. Reduction was 0.85 ± 0.38 mm and 1.02 ± 0.45 mm in ridge width, 0.61 ± 0.20 mm and 0.72 ± 0.27 mm in buccal and 0.66 ± 0.31 mm and 0.56 ± 0.24 mm in lingual ridge height for the AWTG and ADDG group, respectively (p > .05). Histologically, no inflammatory reactions were noticeable and all samples showed new bone formation. Qualitatively, graft-bone amalgamations were more pronounced in ADDG samples. Histomorphometrically, new bone, graft remnants and soft tissue occupied 37.55% ± 8.94%, 17.05% ± 5.58% and 45.4% ± 4.06% of the areas in the AWTG group and 48.4% ± 11.56%, 11.45% ± 4.13% and 40.15% ± 7.73% in the ADDG group of the examined areas, respectively (p > .05)., Conclusions: AWTG and ADDG are similarly effective in ARP. Yet, histologically ADDG seems to demonstrate better graft remodeling, integration and osteoinductive properties., (© 2021 The Authors. Clinical Oral Implants Research published by John Wiley & Sons Ltd.)

Aim: Toll-like receptors are key players in mesenchymal stem/progenitor cells' micro-environmental crosstalk, endorsing various biological reactions. For the first time, this study investigates the effects of TLR3-ligation on gingival mesenchymal stem/progenitor cells (G-MSCs) stemness and differentiation properties., Material and Methods: G-MSCs (n = 5) were isolated, sorted using anti-STRO-1 antibodies,and sowed on culture dishes to generate colony-forming units (CFUs), and their stem/progenitor cells' features and TLR3 expression were characterized. Subsequently, TLR3 activation of G-MSCs via Poly (I:C) was done, followed by an analysis of the expression of pluripotency-related factors, mesenchymal stemness-associated surface markers, and the ability to form CFUs and multilineage differentiation, using qualitative and quantitative histochemistry and RT-PCR., Results: G-MSCs demonstrated all predefined stem/progenitor cells' characteristics and TLR3 expression. TLR3-activated G-MSCs showed a significantly reduced ability to form CFUs and pluripotency transcriptional factors expression. Mesenchymal stem/progenitor cell-associated surface markers and multilinear differentiation potential were significantly higher following TLR3 ligation (p < .05, Wilcoxon signed rank test)., Conclusions: TLR3-mediated activation maintains the mesenchymal stem/progenitor cells phenotype and drives G-MSCs' differentiation and commitment, with a shift away from an undifferentiated pluripotent cellular phenotype. This distinctive modulation could influence potential therapeutic applications of G-MSCs., (© 2020 The Authors. Journal of Clinical Periodontology published by John Wiley & Sons Ltd.)

Great strides have been made in triple negative breast cancer (TNBC) treatment, which represents 20% of total predicted annual US breast cancer (BC) cases. Despite the development of several therapeutics, TNBC patients have poor overall survival rate, compared to other BC patients, justifying the urgent need to discover new entities for use to control TNBC. Chalcones are important natural products with diverse bioactivities including anticancer effects. This study aimed to design, synthesize and validate novel chalcone leads as potential therapies for TNBC. Fourteen novel chalcone analogs were designed and synthesized comprising alicyclic amines (pyrrolidine, morpholine and piperidine) or nitrogen mustard (Bis-(2-chloroethyl) amine) substituents. Among them, compound 14((E)-3-(4-(Bis(2-chloroethyl) amino) phenyl)-1-(3-methoxyphenyl) prop-2-en-1-one) was identified as the most effective against TNBC and other BC phenotypes, with anti-proliferative IC 50 values ranging between 3.94 and 9.22 μM against the TNBC cell lines MDA-MB-231 and MDA-MB-468, as well as against the estrogen positive MCF-7 cell line. Chalcone 14 effectively suppressed the colony formation capacity of MDA-MB-231, MDA-MB-468, and MCF-7 cell lines at 5 and 10 μM treatment concentrations. Furthermore, compound 14 has significantly inhibited cell invasion and migration of MDA-MB-231 and MCF-7 BC cell lines. Additionally, compound 14 had significantly promoted apoptosis by upregulating BAX and downregulating Bcl-2 proteins. Compound 14 induced significant cell cycle arrest of TNBC cells at the G2/M phase. It also induced a reversal of Epithelial Mesenchymal Transition (EMT) by upregulating the epithelial markers E-cadherin and Pan-cadherin and downregulating FAK. Furthermore, it had dramatically diminished new vessel formation (vasculogenesis) in chick chorioallantoic membrane (CAM) model by 60.20 ± 8.47%. Chalcone 14 inhibited 46.41 ± 0.71% of the TNBC MAD-MB-231 cells growth in a nude mouse orthotopic xenograft model in comparison with vehicle control treated animals. Collectively, this study results propose chalcone 14 as a promising lead molecule for the control of TNBC as well as other breast cancer phenotypes., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Epidemiological and clinical studies compellingly showed the ability of Mediterranean diet rich in extra-virgin olive oil (EVOO) to reduce multiple diseases such as cancer, cardiovascular diseases, and aging cognitive functions decline. The S -(-)-Oleocanthal (OC) is a minor phenolic secoiridoid exclusively found in extra-virgin olive oil (EVOO). OC recently gained notable research attention due to its excellent in vitro and in vivo biological effects against multiple cancers, inflammations, and Alzheimer's disease. However, OC safety has not been comprehensively studied yet. This study reports for the first time the detailed safety of oral single OC dose in Swiss albino mice, applying the OECD 420 procedure. Male and female Swiss albino mice ( n = 10) were orally treated with a single OC dose of either 10, 250, or 500 mg/kg bodyweight or equivalent volumes of distilled water. Mice fed a regular diet, and carefully observed for 14 days. Further, mice were then sacrificed, blood samples, and organs were collected and subjected to hematological, biochemical, and histological examinations. OC 10 mg/kg oral dose appears to be without adverse effects. Further, 250 mg/kg OC, p.o., is suggested as a possible upper dose for preclinical studies in the future., Competing Interests: K. El Sayed is a Co-founder and Medicinal Chemistry Chief Scientific Officer without compensation in the Shreveport, Louisiana-based Oleolive.

S -(-)-Oleocanthal (OC), a naturally occurring phenolic secoiridoid exclusively found in extra-virgin olive oil (EVOO), is a potential nutraceutical therapeutic for inflammation, neurodegenerative diseases, and many malignancies, especially breast cancer (BC). The oral delivery of OC is challenging because of its irritative, bitter, and pungent taste and exceptional chemistry, including two reactive aldehydes, phenolic, and ester groups. OC irritation did not correlate with CO 2 -induced irritation, and hence, OC was not exerting generalized acid-sensing irritation. The objective of this study was to develop an effervescent formulation of OC with an effective CO 2 -induced masked taste maintaining the efficacy against the estrogen receptor (ER) and HER2 positive BC. Several ratios of acid and carbonate sources were screened, and five effervescent formulations EF1-EF5 were selected and prepared based on their pH and effervescence time. OC formulations were characterized using differential scanning calorimetry, FT-IR spectroscopy, and scanning electron microscopy analyses. OC formulations exhibited acceptable flowability and effervescence time. Based on physical characteristics and improved OC release, formulation EF-2 was selected for subsequent studies. EF-2 showed effective OC taste masking, as suggested by electronic artificial tongue and mouse preference tests. EF-2 suppressed more than 70% of the hormone and HER2-positive BT-474 BC cell growth in a nude mouse xenograft model. Furthermore, EF-2 demonstrated significant inhibition of BT-474 tumor cell locoregional recurrence after primary tumor surgical excision. EF-2-treated mouse sera had significantly reduced CA 15-3 levels, the human BC recurrence marker, compared to the placebo control group at the end of the study. These results highlight the potential of the OC formulation EF-2 as a prospective nutraceutical for the control and prevention of ER + /HER + BC progression and locoregional recurrence., Competing Interests: K. El Sayed is a Chief Scientific Officer in the Shreveport, Louisiana-based Oleolive without compensations. The rest of coauthors declare no competing financial interest.

Aim: A range of predictors for tooth loss in periodontitis patients have been reported. We performed a systematic review and meta-analysis to assess the consistency and magnitude of any association between a total of 12 predictors and tooth loss., Materials and Methods: Medline/Embase/Central were searched for longitudinal studies investigating the association between predictors and tooth loss in periodontitis patients. Random-effects meta-analysis was performed, and study quality assessed., Results: Twenty studies (15,422 patients, mean follow-up: 12 years) were included. The mean annual tooth loss/patient was 0.12 (min./max: 0.01/0.36). Older patients (n = 8 studies; OR: 1.05, 95% CI: 1.03-1.08/year), non-compliant ones (n = 11; 1.51, 1.06-2.16), diabetics (n = 7; 1.80, 1.26-2.57), those with IL-1-polymorphism (n = 3; 1.80; 1.29-2.52) and smokers (n = 15; 1.98, 1.58-2.48) had a significantly higher risk of tooth loss. Teeth with bone loss (n = 3; 1.04, 1.03-1.05/%), high probing pocket depth (n = 6; 3.19, 1.70-5.98), mobility (n = 4; 3.71, 1.65-8.38) and molars (n = 4; 4.22, 2.12-8.39), especially with furcation involvement (n = 5; 2.68, 1.75-4.08) also showed higher risks. Gender (n = 16; 0.95, 0.86-1.05) and endodontic affection (n = 3; 3.62, 0.99-13.2) were not significantly associated with tooth loss., Conclusions: Older, non-compliant, smoking or diabetic patients, and teeth with bone loss, high probing pocket depth, mobility, or molars, especially with furcation involvement showed higher risks of tooth loss., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The use of quantum dots (QD) in various medical and industrial applications may cause these nanoparticles to leak into waterways and subsequently enter the food chain. Therefore, if we intend to use QD, we must first know their potential environmental implications. In this work, cadmium selenide/zinc sulfide core/shell QD were synthesized, and then, biocompatible, water-dispersed QD were coated with silica (Si-QD). The QD were characterized by X-ray diffraction (XRD), high-resolution transmission electron microscopy (HRTEM) combined with energy-dispersive X-ray spectroscopy (EDX), and UV-Vis absorption analysis, which revealed that these surface-engineered QD have a highly crystalline, homogeneous spherical shape measuring approximately 25 nm. The cytotoxicity of the nanoparticles in the green algae Chlamydomonas reinhardtii was studied by incubating the algae cells with Si-QD and determining the optical density of algal cell culture, cell counts, and cells sizes by microflow cytometry. These measurements indicated that Si-QD are biocompatible up to a concentration of 25 ng/ml. Finally, the cellular uptake of Si-QD into C. reinhardtii was monitored by confocal laser scanning microscopy (CLSM). In conclusion, our results reveal that surface-engineered Cd-QD can penetrate the cells of aquatic organisms, which ensures a serious impact on the food chain and consequently the environment. On the other hand, the results also highlight a new potential method for bioremediation of Cd-QD by green algae, especially C. reinhardtii., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Epidemiological and clinical studies compellingly documented the ability of Mediterranean diet rich in extra-virgin olive oil (EVOO) to reduce breast and colon cancers incidence, cardiovascular diseases, and aging cognitive functions decline. (-)-Oleocanthal (OC) and other EVOO phenolics gain progressive research attention due to their documented biological effects against cancer, inflammations, and Alzheimer's disease. There is no simple, reliable, and cost-effective isolation protocol for EVOO phenolics, which hinder their therapeutic applications. This study develops novel methods to isolate OC and other EVOO phenolics. This includes the use of ultra-freezing to eliminate most EVOO fats and the successful water capacity to efficiently extract OC and EVOO phenolics as self-emulsified nano-emulsion. Subsequent resin entrapment and size exclusion chromatography afforded individual EVOO phenolics in high purity. OC in vitro and in vivo oral anti-breast cancer (BC) activities validated its lead candidacy. Effective isolation of EVOO phenolics provided in this study will facilitate future preclinical and clinical investigations and stimulate the therapeutic development of these important bioactive natural products., Competing Interests: K. El Sayed and A. Kaddoumi are Chief Scientific Officers without compensation in Oleolive. The isolation method reported in this manuscript is a patent pending topic (PCT/US2017/043308,WO2018/017967). This does not alter the authors’ adherence to PLOS ONE policies on sharing data and materials.

We have previously shown in young males that the rate of rise in blood pressure (BP) at the onset of mental stress determines whether or not muscle sympathetic nerve activity (MSNA) has a role in driving the pressor response. The aim of this study was to investigate these interactions in young females. BP and MSNA were recorded continuously in 19 females and 21 males during 2-min mental stressors (mental arithmetic and Stroop test). Physical stressor tasks (cold pressor, handgrip exercise, postexercise ischemia) were also performed. During the first minute of mental arithmetic, the rate of rise in mean arterial pressure (MAP) was significantly greater in negative responders (mean decrease in MSNA) compared with positive responders (mean increase in MSNA) in both males (1.9 ± 0.7 vs. 0.7 ± 0.3 mmHg/sec) and females (1.0 ± 0.3 vs. 0.5 ± 0.2 mmHg/sec). For the Stroop test, there was no significant difference in the rate of the rise in BP between positive and negative responders (P > 0.05). However, peak changes in MAP were significantly greater in negative responders compared with positive responders in both males (22 ± 6 vs. 13 ± 3 mmHg) and females (12 ± 2 vs. 6 ± 1 mmHg). Sympathetic baroreflex sensitivity was greater in negative responders and may contribute to the fall in MSNA experienced by these individuals during mental stress. During physical stressors there were consistent increases in BP and MSNA in males and females. The findings suggest that, in both males and females, BP reactivity at the onset of mental stress dictates whether or not there is an increase or decrease in MSNA., (© 2018 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.)

Polyphenols are known for their antimicrobial activity, whilst both polyphenols and the globular protein β-lactoglobulin (bLG) are suggested to have antioxidant properties and promote cell proliferation. These are potentially useful properties for a tissue-engineered construct, though it is unknown if they are retained when both compounds are used in combination. In this study, a range of different microbes and an osteoblast-like cell line (human fetal osteoblast, hFOB) were used to assess the combined effect of: (1) green tea extract (GTE), rich in the polyphenol epigallocatechin gallate (EGCG), and (2) whey protein isolate (WPI), rich in bLG. It was shown that approximately 20-48% of the EGCG in GTE reacted with WPI. GTE inhibited the growth of Gram-positive bacteria, an effect which was potentiated by the addition of WPI. GTE alone also significantly inhibited the growth of hFOB cells after 1, 4, and 7 days of culture. Alternatively, WPI significantly promoted hFOB cell growth in the absence of GTE and attenuated the effect of GTE at low concentrations (64 µg/mL) after 4 and 7 days. Low concentrations of WPI (50 µg/mL) also promoted the expression of the early osteogenic marker alkaline phosphatase (ALP) by hFOB cells, whereas GTE inhibited ALP activity. Therefore, the antioxidant effects of GTE can be boosted by WPI, but GTE is not suitable to be used as part of a tissue-engineered construct due to its cytotoxic effects which negate any positive effect WPI has on cell proliferation., (© 2019 S. Karger AG, Basel.)

Objective and Methods: Muscle sympathetic nerve activity and baroreflex sensitivity were examined at rest before, during (weeks 6, 11, 17, 22, 25, 33 and 36) and after a normotensive pregnancy., Results: Muscle sympathetic nerve activity is elevated during pregnancy with a large peak in the first trimester (Δ17 bursts/min) and a secondary peak in the third trimester (Δ11 bursts/min). Cardiac baroreflex sensitivity peaked in the first trimester (10 vs. 6 ms/mmHg pre-pregnancy), whereas sympathetic baroreflex sensitivity was greater throughout., Interpretation: The increase in sympathetic outflow early in pregnancy cannot be explained by a reduction in baroreflex sensitivity, while the secondary increase in burst frequency in the third trimester may, in part, be explained by the elevated heart rate.

This study aimed to assess the revascularisation potential of necrotic mature teeth in a dog model following different protocols. Periapical infection was induced in 54 mature premolars. Teeth were distributed into seven groups: (1) Double-antibiotic-paste/Blood clot, (2) Ciprofloxacin/collagen, (3) Double-antibiotic-paste/Collagen, (4) Modified Tri-antibiotic-paste /collagen, (5) Ciprofloxacin/Gelfoam, (6) Double-antibiotic-paste/Gelfoam, and (7) Modified Triantibiotic- paste/Gelfoam. Positive and negative controls included infected and healthy teeth, respectively, (n = 12 roots/group). Canals were apically shaped to size 0.6 mm then disinfected for 1 month. Intra-canal bleeding was induced then scaffolds were applied for another month. Teeth and supporting bone were surgically sampled. Tissues were histologically scored and vimentin immuno-intensity was estimated. Ciprofloxacin and Double-antibiotic paste/Collagen resulted in significantly better corono-apical tissue ingrowths, vascularity, cementum formation and significantly lower inflammatory extents (P < 0.05).These groups also showed significantly higher Vimentin intensities, (P < 0.05). The applied protocols revascularised necrotic mature canals and reduced inflammation particularly in the Ciprofloxacin/collagen and Double-antibiotic-paste/collagen groups., (© 2017 Australian Society of Endodontology Inc.)

To exert anticaries effects, probiotics are described to inhibit growth and biofilm formation of cariogenic bacteria such as Streptococcus mutans (SM). We screened 8 probiotics and assessed how SM growth or biofilm formation inhibition affects cariogenicity of probiotic-SM mixed-species biofilms in vitro. Growth inhibition was assessed by cocultivating probiotics and 2 SM strains (ATCC 20532/25175) on agar. Probiotics were either precultured before SM cultivation (exclusion), or SM precultured prior to probiotic cultivation (displacement). Inhibition of SM culture growth was assessed visually. Inhibition of SM biofilm formation on bovine enamel was assessed using a continuous-flow short-term biofilm model, again in exclusion or displacement mode. The cariogenicity of mixed-species biofilms of SM with the most promising growth and biofilm formation inhibiting probiotic strains was assessed using an artificial mouth model, and enamel mineral loss (ΔZ) was measured microradiographically. We found limited differences in SM growth inhibition in exclusion versus displacement mode, and in inhibition of SM 20532 versus 25175. Results were therefore pooled. Lactobacillus acidophilus LA-5 inhibited significantly more SM culture growth than most other probiotics. L. casei LC-11 inhibited SM biofilm formation similarly to other alternatives but showed the highest retention of probiotics in the biofilms (p < 0.05). Mineral loss from SM monospecies biofilms (ΔZ = 9,772, 25th/75th percentiles: 6,277/13,558 vol% × µm) was significantly lower than from mixed-species SM × LA-5 biofilms (ΔZ = 24,578, 25th/75th percentiles: 19,081/28,768 vol% × µm; p < 0.01) but significantly higher than from SM × LC-11 biofilms (ΔZ = 4,835, 25th/75th percentiles: 263/7,865 vol% × µm; p < 0.05). Probiotics inhibiting SM culture growth do not necessarily reduce the cariogenicity of SM-probiotic biofilms. Nevertheless, SM biofilm formation inhibition may be relevant in the reduction of cariogenicity., (© 2017 S. Karger AG, Basel.)

Key Points: Research indicates that individuals may experience a rise (positive responders) or fall (negative responders) in muscle sympathetic nerve activity (MSNA) during mental stress. In this study, we examined the early blood pressure responses (including the peak, time of peak and rate of rise in blood pressure) to mental stress in positive and negative responders. Negative MSNA responders to mental stress exhibit a more rapid rise in diastolic pressure at the onset of the stressor, suggesting a baroreflex-mediated suppression of MSNA. In positive responders there is a more sluggish rise in blood pressure during mental stress, which appears to be MSNA-driven. This study suggests that whether MSNA has a role in the pressor response is dependent upon the reactivity of blood pressure early in the task., Abstract: Research indicates that individuals may experience a rise (positive responders) or fall (negative responders) in muscle sympathetic nerve activity (MSNA) during mental stress. The aim was to examine the early blood pressure response to stress in positive and negative responders and thus its influence on the direction of change in MSNA. Blood pressure and MSNA were recorded continuously in 21 healthy young males during 2 min mental stressors (mental arithmetic, Stroop test) and physical stressors (cold pressor, handgrip exercise, post-exercise ischaemia). Participants were classified as negative or positive responders according to the direction of the mean change in MSNA during the stressor tasks. The peak changes, time of peak and rate of changes in blood pressure were compared between groups. During mental arithmetic negative responders experienced a significantly greater rate of rise in diastolic blood pressure in the first minute of the task (1.3 ± 0.5 mmHg s -1 ) compared with positive responders (0.4 ± 0.1 mmHg s -1 ; P = 0.03). Similar results were found for the Stroop test. Physical tasks elicited robust parallel increases in blood pressure and MSNA across participants. It is concluded that negative MSNA responders to mental stress exhibit a more rapid rise in diastolic pressure at the onset of the stressor, suggesting a baroreflex-mediated suppression of MSNA. In positive responders there is a more sluggish rise in blood pressure during mental stress, which appears to be MSNA-driven. This study suggests that whether MSNA has a role in the pressor response is dependent upon the reactivity of blood pressure early in the task., (© 2016 The Authors. The Journal of Physiology © 2016 The Physiological Society.)

Low baroreflex sensitivity (BRS) is associated with elevated cardiovascular risk. However, the evidence is based primarily on measurements of cardiac BRS. It cannot be assumed that cardiac or sympathetic BRS alone represent a true reflection of baroreflex control of blood pressure. The aim of this study was to examine the relationship between spontaneous sympathetic and cardiac BRS in healthy, young individuals. Continuous measurements of blood pressure, heart rate, and muscle sympathetic nerve activity (MSNA) were made under resting conditions in 50 healthy individuals (18-28 years). Sympathetic BRS was quantified by plotting MSNA burst incidence against diastolic pressure (sympathetic BRSinc), and by plotting total MSNA against diastolic pressure (sympathetic BRStotal). Cardiac BRS was quantified by plotting R-R interval against systolic pressure using the sequence method. Significant sympathetic BRSinc and cardiac BRS slopes were obtained for 42 participants. A significant positive correlation was found between sympathetic BRSinc and cardiac BRS (r = 0.31, P = 0.049). Among this group, significant sympathetic baroreflex slopes were obtained for 39 participants when plotting total MSNA against diastolic pressure. In this subset, a significant positive correlation was observed between sympathetic BRStotal and cardiac BRS (r = 0.40, P = 0.012). When males and females were assessed separately, these modest relationships only remained significant in females. Analysis by gender revealed correlations in the females between sympathetic BRSinc and cardiac BRS (r = 0.49, P = 0.062), and between sympathetic BRStotal and cardiac BRS (r = 0.57, P = 0.025). These findings suggest that gender interactions exist in baroreflex control of blood pressure, and that cardiac BRS is not appropriate for estimating overall baroreflex function in healthy, young populations. This relationship warrants investigation in aging and clinical populations., (© 2015 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.)

Hypothesizing that the implantation of non-articular (heterotopic) chondrocytes might be an alternative approach to support articular cartilage repair, we analyzed joint cartilage defect healing in the rabbit model after implantation of autologous auricle-derived (auricular) chondrocytes. Autologous lapine articular and auricular chondrocytes were cultured for 3 weeks in polyglycolic acid (PGA) scaffolds before being implanted into critical sized osteochondral defects of the rabbit knee femoropatellar groove. Cell-free PGA scaffolds and empty defects served as controls. Construct quality was determined before implantation and defect healing was monitored after 6 and 12 weeks using vitality assays, macroscopical and histological score systems. Neo-cartilage was formed in the PGA constructs seeded with both articular and auricular chondrocytes in vitro and in vivo. At the histological level, cartilage repair was slightly improved when using autologous articular chondrocyte seeded constructs compared to empty defects and was significantly superior compared to defects treated with auricular chondrocytes 6 weeks after implantation. Although only the immunohistological differences were significant, auricular chondrocyte implantation induced an inferior healing response compared with the empty defects. Elastic auricular chondrocytes might maintain some tissue-specific characteristics when implanted into joint cartilage defects which limit its repair capacity., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

In response to low oxygen supply, cancer cells elevate production of HIF-1α, a hypoxia-inducible transcription factor that subsequently acts to stimulate blood vessel formation and promote survival. Studies were conducted to determine the role of δ-tocotrienol and a semisynthetic δ-tocotrienol oxazine derivative, compound 44, on +SA mammary tumor cell hypoxic response. Treatment with 150 µM CoCl2 induced a hypoxic response in +SA mammary tumor cells as evidenced by a large increase in HIF-1α levels, and combined treatment with compound 44 attenuated this response. CoCl2-induced hypoxia was also associated with a large increase in Akt/mTOR signaling, activation of downstream targets p70S6K and eIF-4E1, and a significant increase in VEGF production, and combined treatment with compound 44 blocked this response. Additional in vivo studies showed that intralesional treatment with compound 44 in BALB/c mice bearing +SA mammary tumors significantly decreased the levels of HIF-1α, and this effect was associated with a corresponding decrease in Akt/mTOR signaling and activation of downstream targets p70S6 kinase and eIF-4E1. These findings demonstrate that treatment with the δ-tocotrienol oxazine derivative, compound 44, significantly attenuates +SA mammary tumor cell compensatory responses to hypoxia and suggests that this compound may provide benefit in the treatment of rapidly growing solid breast tumors.

Minipigs are widely used as a large animal model for cartilage repair. However, many in vitro studies are based on porcine chondrocytes derived from abundantly available premature hybrid pigs. It remains unclear whether pig line-dependent differences exist which could limit the comparability between in vitro and in vivo results using either hybrid or miniature pig articular chondrocytes. Porcine knee joint femoral cartilage was isolated from 3- to 5-month-old hybrid pigs and Göttingen minipigs. Cartilage from both pig lines was analysed for thickness, zonality, cell content, size and proteoglycan deposition. Cultured articular chondrocytes from both pig lines were investigated for gene and/or protein expression of cartilage-specific proteins such as type II collagen, aggrecan, the chondrogenic transcription factor Sox9, non-specific type I collagen and the cell-matrix receptor β1-integrin. Cartilage was significantly thinner in the miniature pig compared to the hybrid pig, but the differences between the medial and lateral femur condyles did not reach a significant level. Knee joint cartilage zone formation started only in the minipig, whereas cellularity and cell diameters were comparable in both pig lines. Blood vessels could be detected in the hybrid pig but not the minipig cartilage. Sulphated proteoglycan deposition was more pronounced in cartilage zones II-IV of both pig lines. Minipig chondrocytes expressed type II and I collagen, Sox9 and β1-integrin at a higher level than hybrid pig chondrocytes. These distinct line-dependent differences should be considered when using hybrid pig-derived chondrocytes for tissue engineering and Göttingen minipigs as a large animal model.

Implantation of non-articular (heterotopic) chondrocyte-based implants might be an alternative approach to articular cartilage repair. This strategy could be helpful in cases in which there are no or too few articular chondrocytes available. Therefore, this study was undertaken to compare joint cartilage defect healing in the minipig model after implantation of heterotopic auricular and orthotopic articular chondrocytes. Poly-glycolic acid (PGA) associated three-dimensional (3D) constructs were prepared culturing autologous minipig-derived articular and auricular chondrocytes for 7 days in a dynamic culture system. Chondrocyte PGA constructs were implanted into 8mm diameter and ∼1.1mm deep chondral defects within the medial and lateral condyles of the minipig knee joints. Empty defects served as controls for assessment of the intrinsic healing response. Defect healing was monitored 6 months post implantation using a macroscopic and microscopic score system and biomechanical analysis. Neo-cartilage formation could be observed in the PGA constructs seeded with articular and auricular chondrocytes in vivo. The defect healing did not significantly differ at the macroscopic and histological level in response to implantation of either autologous articular or auricular chondrocytes seeded constructs compared with the empty defects. Although the differences were not significant, the auricular chondrocytes-based implants led to a slightly inferior repair quality at the macroscopic level, but a histologically superior healing response when compared with the empty defect group. However, biomechanical analysis revealed a higher stiffness in repair tissues produced by auricular chondrocyte implantation compared with the other groups. Deduced from these results, articular chondrocytes represent the preferable cell source for implantation., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

The study examined the effects of larval trematode infection on the neutral lipid and phospholipid content of Biomphalaria alexandrina infected with Schistosoma mansoni. Uninfected snails were used as matched controls. As determined by qualitative high-performance silica gel thin-layer chromatography (HPTLC), the major neutral lipids present in the whole bodies and digestive gland-gonad complexes in both infected and uninfected snail populations were free sterols, free fatty acids, and triacylglycerols, and the major polar lipids were phosphatidylcho-line and phosphatidyl ethanolamine. Quantitative analysis by HPTLC with visible and UV scanning reflectance densitometry showed the snail's digestive gland lipid level was found to be almost halved in 20 days post infection; a more then 80% reduction being visible after the subsequent 40 and 60 days.

Tissue Engineering is an important method for generating cartilage tissue with isolated autologous cells and the support of biomaterials. In contrast to various gel-like biomaterials, human demineralized bone matrix (DBM) guarantees some biomechanical stability for an application in biomechanically loaded regions. The present study combined for the first time the method of seeding chondrocyte-macroaggregates in DBM for the purpose of cartilage tissue engineering. After isolating human nasal chondrocytes and creating a three-dimensional macroaggregate arrangement, the DBM was cultivated in vitro with the macroaggregates. The interaction of the cells within the DBM was analyzed with respect to cell differentiation and the inhibitory effects of chondrocyte proliferation. In contrast to chondrocyte-macroaggregates in the cell-DBM constructs, morphologically modified cells expressing type I collagen dominated. The redifferentiation of chondrocytes, characterized by the expression of type II collagen, was only found in low amounts in the cell-DBM constructs. Furthermore, caspase 3, a marker for apoptosis, was detected in the chondrocyte-DBM constructs. In another experimental setting, the vitality of chondrocytes as related to culture time and the amount of DBM was analyzed with the BrdU assay. Higher amounts of DBM tended to result in significantly higher proliferation rates of the cells within the first 48 h. After 96 h, the vitality decreased in a dose-dependent fashion. In conclusion, this study provides the proof of concept of chondrocyte-macroaggregates with DBM as an interesting method for the tissue engineering of cartilage. The as-yet insufficient redifferentiation of the chondrocytes and the sporadic initiation of apoptosis will require further investigations.

Tissue trauma induces an inflammatory response associated with a cytokine release that may engage complement pathways. Cytokine-mediated complement expression may contribute to cartilage degradation. Hence, we analysed the complement expression profile in primary articular and non-articular chondrocytes and its interrelation with cytokines. The expression of the anaphylatoxin receptors (C3aR and C5aR) and the complement regulatory proteins (CPRs) CD35, CD46, CD55 and CD59 was studied in cultured articular, auricular and nasoseptal chondrocytes using RTD-PCR and immunofluorescence labelling. The complement profile of peripheral blood mononuclear cells (PBMCs) was opposed to the expression in articular chondrocytes. The time-dependent regulation (6 and 24 h) of these complement factors was assessed in articular chondrocytes in response to the cytokines TNFα, IL-10 or TNFα combined with IL-10 (each 10 ng/mL). C3aR, C5aR, CD46, CD55 and CD59 but almost no CD35 mRNA was expressed in any of chondrocyte types studied. The anaphylatoxin receptor expression was lower and that of the CRPs was higher in chondrocytes when compared with PBMCs. The majority of the studied complement factors were expressed at a significantly lower level in non-articular chondrocytes compared with the articular chondrocytes. TNFα significantly increased the C3aR expression in chondrocytes after 6 and 24 h. TNFα + IL-10 significantly downregulated C5aR and IL-10 significantly inhibited the CD46 and CD55 gene expression after 24 h. C5aR and CD55 could be localised in cartilage in situ. Anaphylatoxin receptors and CRPs are regulated differentially by TNFα and IL-10. Whether cytokine-induced complement activation occurs in response to cartilage trauma has to be further identified.

The resistance of tumor cells to a broad range of anticancer agents continues to be a problem for the success of cancer chemotherapy. Multidrug resistance (MDR) is due in part to three drug transporter proteins: ABCB1/P-glycoprotein (P-gp), ABCC1/multidrug resistance protein 1 (MRP1) and ABCG2/breast cancer resistance protein (BCRP). These transporters are part of the ATP-binding cassette (ABC) superfamily, whose members function as ATP-dependent drug-efflux pumps. Their activity can be blocked by various drugs such as verapamil (calcium channel blocker) and cyclosporin A (immunosuppressive agent), etc. These compounds are called MDR modulators or reversals. This review highlights several marine natural products with reversal effect on multidrug resistance in cancer, including agosterol A, ecteinascidin 743, sipholane triterpenoids, bryostatin 1, and welwitindolinones.

Using low-frequency (0.08-0.18 Hz) sinusoidal galvanic vestibular stimulation (sGVS), we recently showed that two peaks of modulation of muscle sympathetic nerve activity (MSNA) and skin sympathetic nerve activity (SSNA) occurred for each cycle of stimulation: a large peak associated with the positive peak of the sinusoid (defined as the primary peak) and a smaller peak (defined as the secondary peak) related to the negative peak of the sinusoid. However, these recordings were only made from the left common peroneal nerve, so to investigate lateralisation of vestibulosympathetic reflexes, concurrent recordings were made from both sides of the body. Tungsten microelectrodes were inserted into muscle or cutaneous fascicles of the left and right common peroneal nerves in 17 healthy individuals. Bipolar binaural sinusoidal GVS (±2 mA, 100 cycles) was applied to the mastoid processes at 0.08 Hz. Cross-correlation analysis revealed that vestibular modulation of MSNA (10 bilateral recordings) and SSNA (6 bilateral recordings) on the left side was expressed as a primary peak related to the positive phase of the sinusoid and a secondary peak related to the negative phase of the sinusoid. Conversely, on the right side, the primary and secondary peaks were reversed: the secondary peak on the right coincided with the primary peak on the left and vice versa. Moreover, differences in pattern of outflow were apparent across sides. We believe the results support the conclusion that the left and right vestibular nuclei send both an ipsilateral and contralateral projection to the left and right medullary output nuclei from which MSNA and SSNA originate. This causes a "flip-flop" patterning between the two sympathetic outflows: when vestibular modulation of a burst is high on the left, it is low on the right, and when modulation is low on the left, it is high on the right.

Aim: The aim of this study was to determine the survival rates of questionable and hopeless teeth in patients with aggressive periodontitis (AgP) and chronic periodontitis (CP) during 15 years of supportive periodontal therapy (SPT)., Materials and Methods: Thirty-four AgP and 34 CP patients (SPT≥10 years) with bone loss of ≥50% at ≥2 teeth were consecutively recruited. Bone loss was measured on digitized radiographs and teeth were categorized as "questionable" (≥50 to <70% bone loss) or as "hopeless" (≥70%). Progression in pocket probing depths (PPD) during SPT, tooth loss and reasons for extraction were analysed., Results: In AgP patients, 262 teeth were considered as questionable and 63 as hopeless (CP: 149/51). During active periodontal therapy, 25 questionable and 26 hopeless teeth were extracted (CP: 12/16). During 15.3 ± 4.1 years of SPT of AgP 28 questionable and 15 hopeless teeth were removed (CP: 28/12). The mean tooth loss per patient during SPT in total was 0.14 (AgP) and 0.16 (CP) teeth/year. There were no significant differences in tooth loss or longitudinal progression of PPD between AgP and CP patients., Conclusions: In patients with AgP, 88.2% (209 of 237) of questionable and 59.5% (22 of 37) of hopeless teeth survived 15 years during regular SPT in a dental school department., (© 2011 John Wiley & Sons A/S.)

Injured articular cartilage is limited in its capacity to heal. Autologous chondrocyte transplantation (ACT) is a suitable technique for cartilage repair, but it requires articular cartilage biopsies for sufficient autologous chondrocyte expansion in vitro. Hence, ACT is restricted by donor-site morbidity and autologous articular chondrocytes availability. The use of nonarticular heterotopic chondrocytes such as auricular, nasoseptal, or costal chondrocytes for ACT might overcome these limitations: heterotopic sources show lesser donor-site morbidity and a comparable extracellular cartilage matrix synthesis profile to articular cartilage. However, heterotopic (h)ACT poses a challenge. Particular tissue characteristics of heterotopic cartilage, divergent culturing peculiarities of heterotopic chondrocytes, and the advantages and drawbacks related to these diverse cartilage sources were critically discussed. Finally, available in vitro and in vivo experimental (h)ACT approaches were summarized. The quality of the cartilage engineered using heterotopic chondrocytes remains partly controversy due to the divergent methodologies and culture conditions used. While some encouraging in vivo results using (h)ACT have been demonstrated, standardized culturing protocols are strongly required. However, whether heterotopic chondrocytes implanted into joint cartilage defects maintain their particular tissue properties or can be adapted via tissue engineering strategies to fulfill regular articular cartilage functions requires further studies.

Previously, we reported sipholenol A, a sipholane triterpenoid from the Red Sea sponge Callyspongia siphonella, as a potent reversal of multidrug resistance (MDR) in cancer cells that overexpressed P-glycoprotein (P-gp). Through extensive screening of several related sipholane triterpenoids that have been isolated from the same sponge, we identified sipholenone E, sipholenol L and siphonellinol D as potent reversals of MDR in cancer cells. These compounds enhanced the cytotoxicity of several P-gp substrate anticancer drugs, including colchicine, vinblastine and paclitaxel, and significantly reversed the MDR-phenotype in P-gp-overexpressing MDR cancer cells KB-C2 in a dose-dependent manner. Moreover, these three sipholanes had no effect on the response to cytotoxic agents in cells lacking P-gp expression or expressing MRP1 (ABCC1) or MRP7 (ABCC10) or breast cancer resistance protein (BCRP/ABCG2). All three sipholanes (IC(50) >50 μM) were not toxic to all the cell lines that were used. [(3)H]-Paclitaxel accumulation and efflux studies demonstrated that all three triterpenoids time-dependently increased the intracellular accumulation of [(3)H]-paclitaxel by directly inhibiting P-gp-mediated drug efflux. Sipholanes also inhibited calcein-AM transport from P-gp-overexpressing cells. The Western blot analysis revealed that these three triterpenoids did not alter the expression of P-gp. However, they stimulated P-gp ATPase activity in a concentration-dependent manner and inhibited the photolabeling of this transporter with its transport substrate [(125)I]-iodoarylazidoprazosin. In silico molecular docking aided the virtual identification of ligand binding sites of these compounds. In conclusion, sipholane triterpenoids efficiently inhibit the function of P-gp through direct interactions and may represent potential reversal agents for the treatment of MDR., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Alveolar bone grafting is a standard method for treating alveolar cleft. To ensure the best outcome, improving the arch form as well as soft tissue quality in the area around the cleft is recommended. In this study, 11 patients who presented with alveolar cleft and collapsed maxillary arch were treated in the following sequence: transpalatal distraction osteogenesis followed by soft tissue surgery in some cases and by cancellous bone graft. In all cases, transpalatal distraction osteogenesis successfully corrected the transverse maxillary deficiency. One case showed a complete loss of the bone graft. Other minor complications were reported but they did not affect the final outcome., (Copyright 2010 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.)

Nanostructured lipid carriers (NLCs), made from mixtures of solid and liquid lipids, were postulated to have superior properties over solid lipid nanoparticles (SLNs). Nonetheless, the architecture of their inner cores remains elusive. The objective of this study was to elucidate the mode by which tocotrienol-rich fraction (TRF) is entrapped within NLCs and the impact of TRF interaction with solid lipids on the long-term stability of the nanoparticles. The mode of TRF localization was postulated from TEM image analysis and (1)H NMR signals' amplitude. The size, polydispersity, and fusion enthalpy were found to decrease with an increase in TRF loading, which implied a distortion in the crystallinity of the nanoparticles and the preferential entrapment of TRF within the cores of the NLCs. Nonetheless, (1)H NMR spectra of TRF-NLCs broadened as TRF load decreased from 100 to 10%, which was attributed to partial TRF mobility on the surface of the nanoparticles. This was confirmed by TEM images of NLCs at 50% TRF loads. These data led to the conclusion that NLCs have limited capacity to accommodate TRF with the excess being expelled to the surface of the nanoparticles. Such arrangement may have implication on future utility of the NLCs as drug delivery vehicles., (Copyright 2010 Elsevier B.V. All rights reserved.)

Cartilage injury remains a challenge in orthopedic surgery as articular cartilage only has a limited capacity for intrinsic healing. Autologous chondrocyte transplantation (ACT) is a suitable technique for cartilage repair, but requires articular cartilage biopsies for autologous chondrocyte expansion. The use of heterotopic chondrocytes derived from non-articular cartilage sources such as auricular chondrocytes may be a novel approach for ACT. The aim of the study is to evaluate whether co-cultured articular/auricular chondrocytes exhibit characteristics comparable to articular chondrocytes. Analysis of the proliferation rate, extracellular cartilage matrix (ECM) gene and protein expression (type II and I collagen, elastin, lubricin), beta1-integrins and the chondrogenic transcription factor sox9 in articular/auricular chondrocytes was performed using RTD-PCR, flow cytometry, immunofluorescence microscopy and Western blot analysis. Additionally, three-dimensional (3D) chondrocyte mono- and co-cultures were established. The proliferative activity and elastin gene expression were lower and that of type II collagen and lubricin was higher in articular compared with auricular chondrocytes. The species generally did not influence the chondrocyte characteristics, with the exception of type I collagen and sox9 expression, which was higher in porcine but not in human articular chondrocytes compared with both types of auricular chondrocytes. beta1-integrin gene expression did not differ significantly between the chondrocyte types. The type II collagen gene and protein expression was higher in articular chondrocyte monocultures and was slightly higher in co-cultures compared with monocultured auricular chondrocytes. Both chondrocyte types survived in co-culture. Despite their differing expression profiles, co-cultures revealed some adjustment in the ECM expression of both chondrocyte types.

Background: Cytokines are proposed to play important roles in brain tumor biology as well as neurodegeneration or impaired neuronal function., Objectives: This work aimed to check the association of polymorphisms of cytokine genes in Egyptian cases with brain tumors., Methods: This work included 45 cases affected by brain tumors diagnosed as 24 benign and 21 malignant. Their median age was 45 years, and they were 20 males and 25 females. These cases were taken randomly from the Neurosurgery Department of Mansoura University Hospital, Egypt. Case genotypes were compared to 98 healthy unrelated controls from the same locality. DNA was amplified using PCR utilizing sequence specific primers (SSP) for detection of polymorphisms related to TNF-a-308 (G/A), IL-10-1082 (G/A), IL-6-174 (G/C) and IL-1Ra (VNTR) genes., Results: Cases affected with benign brain tumors showed a significant higher frequency of IL-10-1082 A/A [odds ratio (OR=8.0), p<0.001] and IL-6-174 C/C (OR=6.3, p=0.002) homozygous genotypes as compared to controls. Malignant cases, on the other hand, showed significantly higher frequency of IL-6-174 C/C (OR =4.8, p=0.002) homozygous genotype and TNF-a-308 A/A (OR=4.9, p<0.001) homozygous genotype when compared to controls. In the meantime, all cases showed no significant difference regarding the distribution of IL-1Ra VNTR genotype polymorphism compared to controls., Conclusions: Cytokine gene polymorphisms showed a pattern of association with brain tumors which may have potential impact on family counseling and disease management.

Expression of calcitonin (CT) and its receptor (CTR) is elevated in advanced prostate cancer, and activated CT-CTR autocrine axis plays a pivotal role in tumorigenicity and metastatic potential of multiple prostate cancer cell lines. Recent studies suggest that CT promotes prostate cancer metastasis by reducing cell-cell adhesion through the disassembly of tight and adherens junctions and activation of beta-catenin signaling. We attempted to identify a class of molecules that enhances cell-cell adhesion of prostate cells and reverses the disruptive actions of CT on tight and adherens junctions. Screening several compounds led to the emergence of phenyl-methylene hydantoin (PMH) as a lead candidate that can augment cell-cell adhesion and abolish disruptive actions of CT on junctional complexes. PMH reduced invasiveness of PC-3M cells and abolished proinvasive actions of CT. Importantly, PMH did not display significant cytotoxicity on PC-3M cells at the tested doses. I.p. administered PMH and its S-ethyl derivative remarkably decreased orthotopic tumor growth and inhibited the formation of tumor micrometastases in distant organs of nude mice. PMH treatment also reduced the growth of spontaneous tumors in LPB-Tag mice to a significant extent without any obvious cytotoxic effects. By virtue of its ability to stabilize cell junctions, PMH could reverse the effect of CT on junctional disruption and metastasis, which strengthens the possibility of using PMH as a potential drug candidate for CT-positive androgen-independent prostate cancers.

12 patients presenting with long standing temporomandibular joint (TMJ) ankylosis were treated with a costochondral graft inserted through a modified approach. The age of the patients ranged from 5 to 17 years. A preauricular incision was made for resection of the ankylosed condyle. After release of the ankylosis the contralateral rib was harvested with costal cartilage. An intra-oral incision was made along the external oblique ridge to the mucobuccal fold and was used for resection of the coronoid process and insertion and fixation of the graft. The graft was fixed with a minimum of three titanium screws. The patients were instructed to start physiotherapy 1 week postoperatively and were followed up clinically and radiographically using 3D CT. Postoperative results were encouraging, the graft took well in all patients without postoperative infection or graft rejection. The graft was properly positioned in all cases. There were no visible scars as the preauricular scar is relatively hidden, no possibility of damaging the facial nerve or the marginal mandibular branch and shorter operating time.

Through extensive screening of marine sponge compounds, the authors have found that sipholenol A, a sipholane triterpene isolated from the Red Sea sponge, Callyspongia siphonella, potently reversed multidrug resistance (MDR) in cancer cells that overexpressed P-glycoprotein (P-gp). In experiments, sipholenol A potentiated the cytotoxicity of several P-gp substrate anticancer drugs, including colchicine, vinblastine, and paclitaxel, but not the non-P-gp substrate cisplatin, and significantly reversed the MDR of cancer cells KB-C2 and KB-V1 in a concentration-dependent manner. Furthermore, sipholenol A had no effect on the response to cytotoxic agents in cells lacking P-gp expression or expressing MDR protein 1 or breast cancer resistance protein. Sipholenol A (IC(50) > 50 microM) is not toxic to all the cell lines that were used, regardless of their membrane transporter status. Accumulation and efflux studies with the P-gp substrate [(3)H]-paclitaxel demonstrated that sipholenol A time-dependently increased the intracellular accumulation of [(3)H]-paclitaxel by directly inhibiting P-gp-mediated drug efflux. In addition, sipholenol A did not alter the expression of P-gp after treating KB-C2 and KB-V1 cells for 36 h and 72 h. However, it efficaciously stimulated the activity of ATPase of P-gp and inhibited the photolabeling of this transporter with its transport substrate [(125)I]-iodoarylazidoprazosin. Overall, the present results indicate that sipholenol A efficiently inhibits the function of P-gp through direct interactions, and sipholane triterpenes are a new class of potential reversing agents for treatment of MDR in P-gp-overexpressing tumors.