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Recently, new centrifugation protocols for the preparation of platelet-rich fibrin (PRF) have been introduced in an attempt to further improve the beneficial impact of these 2nd generation platelet concentrate membranes. This in-vitro study aimed to compare the biological and physical characteristics of three types of PRF membranes using two different centrifuges with adapted relative centrifugal forces (RCF): leucocyte- and platelet-rich fibrin, advanced platelet-rich fibrin, and advanced platelet-rich fibrin+. Release of growth factors, macroscopic dimensions, cellular content and mechanical properties of the respective membranes, prepared from blood of the same individual were explored. Furthermore, the impact of timing (blood draw-centrifugation and centrifugation-membrane preparation) was assessed morphologically as well as by electron microscopy scanning. No statistically significant differences amongst the three PRF modifications could be observed, neither in their release of growth factors or the cellular content, nor in clot/membrane dimensions. The difference between both centrifuges were negligible when the same g-force was used. A lower g-force, however, reduced membrane tensile strength. Timing in the preparation process had a significant impact. Adaptation of RCF only had a minimal impact on the final characteristics of PRF membranes.

OBJECTIVE AND BACKGROUND: Little is known about structural and mechanical properties of leukocyte- and platelet-rich fibrin (L-PRF) membranes and even less about the influence of antithrombotic drugs on L-PRF. The aim of this in vitro study is therefore to investigate mechanical properties, fibrin structure and cell content of L-PRF membranes and the impact of anticoagulant therapy on L-PRF. METHODS: Blood samples were obtained from 12 volunteers and supplemented with either no, 1.25 IU, 2.5 IU, 5 IU, or 10 IU enoxaparin. L-PRF membranes were characterized with tensile testing, scanning electron microscopy, and measurement of platelets and leukocytes. Control and enoxaparin-supplemented L-PRF membranes were compared. RESULTS: At 10 IU enoxaparin, no L-PRF membranes could be generated, whereas the low doses of 1.25 and 2.5 IU had no influence on L-PRF properties. The mechanical properties, fibrin networks, and number of platelets and leukocytes of 5 IU supplemented membranes were unlike the control membranes, but were not found to be significantly different because of limited sampling and inter- and intra-variability. CONCLUSION: Low doses of the anticoagulant enoxaparin do not affect mechanical properties, fibrin network, nor cellular content of L-PRF, whereas high doses impair L-PRF generation. ispartof: JOURNAL OF PERIODONTAL RESEARCH vol:55 issue:5 pages:686-693 ispartof: location:United States status: published

Leucocyte- and platelet rich fibrin (L-PRF) is an autologous biomaterial used in regenerative procedures. It has an antimicrobial activity against P. gingivalis although the mechanism is not fully understood. It was hypothesized that L-PRF exudate releases hydrogen peroxide and antimicrobial peptides that inhibit P. gingivalis growth. Agar plate and planktonic culture experiments showed that the antimicrobial effect of L-PRF exudate against P. gingivalis was supressed by peroxidase or pepsin exposure. In developing multi-species biofilms, the antimicrobial effect of L-PRF exudate was blocked only by peroxidase, increasing P. gingivalis growth with 1.3 log genome equivalents. However, no effect was shown on other bacteria. Pre-formed multi-species biofilm trials showed no antimicrobial effect of L-PRF exudate against P. gingivalis or other species. Our findings showed that L-PRF exudate may release peroxide and peptides, which may be responsible for its antimicrobial effect against P. gingivalis. In addition, L-PRF exudate had an antimicrobial effect against P. gingivalis in an in vitro developing multi-species biofilm.

Translation is a fundamental step in gene expression that regulates multiple developmental and stress responses. One key step of translation initiation is the association between eIF4E and eIF4G. This process is regulated in different eukaryotes by proteins that bind to eIF4E; however, evidence of eIF4E-interacting proteins able to regulate translation is missing in plants. Here, we report the discovery of CERES, a plant eIF4E-interacting protein. CERES contains an LRR domain and a canonical eIF4E-binding site. Although the CERES–eIF4E complex does not include eIF4G, CERES forms part of cap-binding complexes, interacts with eIF4A, PABP and eIF3, and co-sediments with translation initiation complexes in vivo. Moreover, CERES promotes translation in vitro and general translation in vivo, while it modulates the translation of specific mRNAs related to light and carbohydrate response. These data suggest that CERES is a non-canonical translation initiation factor that modulates translation in plants. Eukaryotic translation initiation factor 4E-interacting proteins able to regulate translation have not yet been reported in plants. A study now characterizes an eIF4E-interacting protein, CERES, as a non-canonical translation initiation factor that modulates translation in plants.

Various studies have described the biological properties of the Leucocyte- and Platelet Rich Fibrin (L-PRF) such as the antimicrobial effect against wound bacteria, but less is known about the effect against periodontal pathogens. The aim of this study was to evaluate the antibacterial properties of the L-PRF membrane and L-PRF exudate against the main periopathogens cultured on agar plates and in planktonic solution. This study demonstrated the antibacterial effect of the L-PRF membrane against P. intermedia, F. nucleatum, and A. actinomycetemcomitans, but especially against P. gingivalis. The L-PRF exudate also showed a strong inhibition against P. gingivalis on agar plates. No inhibition could be observed for the other bacterial strains. Moreover, L-PRF exudate decreased the number of viable P.gingivalis in a planktonic solution in a dose-dependent way. However, A. actinomycetemcomitans showed an increased growth in planktonic solution when in contact with the L-PRF exudate.

AIM: To evaluate dimensional changes in the alveolar ridge and bone structure after tooth extraction when L-PRF or A-PRF+ was used in comparison to unassisted socket healing. MATERIALS AND METHODS: Twenty patients in need of at least three tooth extractions in the aesthetic zone were included. L-PRF, A-PRF+ or control was randomly assigned, leaving one empty socket/edentulous site between conditions. CBCT scans were obtained immediately after tooth extraction and after 3 months of healing. Horizontal and vertical dimensional changes of the ridge and socket fill were calculated. Histological and micro-CT analysis of bone biopsies were used to evaluate post-surgical bone structural healing. RESULTS: Mean horizontal and vertical changes at 1-mm below the crest (buccal and palatal side) were similar for the three sites (p > 0.05). For the socket fill, L-PRF (85.2%) and A-PRF+ (83.8%) showed superior values than the control (67.9%). The histological and radiological analysis reported more newly formed bone for the PRF groups, without any significant differences between both. CONCLUSIONS: PRF matrices failed to reduce the dimensional changes after multiple tooth extractions in the premaxilla. After 3-month healing, both PRF matrices showed radiographically a significant superiority for the socket fill. Histologically, they seemed to accelerate new bone formation. ispartof: JOURNAL OF CLINICAL PERIODONTOLOGY vol:48 issue:7 pages:984-995 ispartof: location:United States status: published

BACKGROUND: This study aims to evaluate the use of the leukocyte- and platelet-rich fibrin (L-PRF) membranes in increasing the width of the keratinized mucosa (KM) around implants. MATERIAL AND METHODS: Eight patients in need for bilateral widening of the KM around implants in the lower jaw were recruited for a spit-mouth randomized controlled trial. At the control site, a free gingival graft (FGG) was used, whereas at the other side (test), L-PRF membranes were applied. The primary outcome was the increase in width of KM around the implants. As secondary outcomes, the postoperative pain and surgical time were assessed. The follow-up period was 6 weeks. RESULTS: A significant increase in the total bucco-lingual width of KM in both groups was observed, with 1.3 mm ± 0.9 extra gain (P

PURPOSE: The leukocyte- and platelet-rich fibrin block (L-PRF block) is a composite graft that combines a xenograft that is acting as a scaffold with L-PRF membranes that serve as a bioactive nodule with osteoinductive capacity. This study evaluated the properties of the L-PRF block and its components in terms of release of growth factors, cellular content, and structure. MATERIALS AND METHODS: The concentration of transforming growth factor-β1 (TGF-β1), vascular endothelial growth factor (VEGF), platelet-derived growth factor-AB (PDGF-AB) and bone morphogenetic protein-1 (BMP-1) released by a L-PRF membrane (mb) and a L-PRF block were examined with ELISA for five time intervals (0 to 4 hours, 4 hours to 1 day, 1 to 3 days, 3 to 7 days, 7 to 14 days). Those levels in L-PRF exudate and liquid fibrinogen were also evaluated. The cellular content of the liquid fibrinogen, L-PRF membrane and exudate was calculated. The L-PRF block was also analyzed by means of a microCT scan and scanning electron microscopy (SEM). RESULTS: TGF-β1 was the most released growth factor after 14 days, followed by PDGF-AB, VEGF, and BMP-1. All L-PRF blocks constantly released the four growth factors up to 14 days. L-PRF membrane and liquid fibrinogen presented high concentration of leukocytes and platelets. The microCT and SEM images revealed the bone substitute particles surrounded by platelets and leukocytes, embedded in a dens fibrin network. CONCLUSION: The L-PRF block consists of deproteinized bovine bone mineral particles surrounded by platelets and leukocytes, embedded in a fibrin network that releases growth factors up to 14 days. ispartof: INTERNATIONAL JOURNAL OF ORAL & MAXILLOFACIAL IMPLANTS vol:34 issue:4 pages:855-864 ispartof: location:United States status: published

Aim To analyse the effect of Leucocyte- and Platelet Rich Fibrin (L-PRF) on bone regeneration procedures and osseointegration. Materials and Methods An electronic and hand search was conducted in three databases (MEDLINE, EMBASE and Cochrane). Only randomised clinical trials, written in English where L-PRF was applied in bone regeneration and implant procedures, were selected. No follow-up restrictions were applied. Results A total of 14 articles were included and processed. Three subgroups were created depending on the application: sinus floor elevation (SFE), alveolar ridge preservation, and implant therapy. In SFE, for a lateral window as well as for the trans-alveolar technique, histologically faster bone healing was reported when L-PRF was added to most common xenografts. L-PRF alone improved the preservation of the alveolar width, resulting in less buccal bone resorption compared to natural healing. In implant therapy, better implant stability over time and less marginal bone loss were observed when L-PRF was applied. Meta-analyses could not be performed due to the heterogeneity of the data. Conclusions Despite the lack of strong evidence found in this systematic review, L-PRF might have a positive effect on bone regeneration and osseointegration. This article is protected by copyright. All rights reserved.

Aim To analyse the regenerative potential of leucocyte‐ and platelet‐rich fibrin (L‐PRF) during periodontal surgery. Materials and Methods An electronic and hand search were conducted in three databases. Only randomized clinical trials were selected and no follow‐up limitation was applied. Pocket depth (PD), clinical attachment level (CAL), bone fill, keratinized tissue width (KTW), recession reduction and root coverage (%) were considered as outcome. When possible, meta‐analysis was performed. Results Twenty‐four articles fulfilled the inclusion and exclusion criteria. Three subgroups were created: intra‐bony defects (IBDs), furcation defects and periodontal plastic surgery. Meta‐analysis was performed in all the subgroups. Significant PD reduction (1.1 ± 0.5 mm, p 0.05), CAL gain (0.2 ± 0.5 mm, p > 0.05), KTW (0.3 ± 0.4 mm, p > 0.05) and recession reduction (0.2 ± 0.3 mm, p > 0.05). Conclusions L‐PRF enhances periodontal wound healing.

This study sought to improve the mechanical and blood-absorbing properties of collagen sponges, while keeping them compressible, by incorporating blended hydroxyapatite (HA)-starch. Results were compared with CollaPlug(®) (pure collagen). The elastic modulus increased from 1.5±0.2kPa for CollaPlug(®) to 49±8kPa for sponges with composition 1:4:10 (collagen:HA:starch, by weight). The modified microstructure and surface area provided by the starch granules on the sponges improved cell viability. Sponges with composition 1:4:10 maintained their blood-clotting capability with almost no change from 5 to 15min after contact with blood, while CollaPlug(®) diminished to about half its capacity to absorb blood and form clots. Incorporation of HA-starch into the sponges with composition of 1:4:10, increased the elastic modulus of the collagen-HA sponges, making them more structurally robust. The viability of cells and the blood-clotting capability increased with starch incorporation.

In many plant species, an exposure to a sublethal temperature triggers an adaptative response called acclimation. This response involves an extensive molecular reprogramming that allows the plant to further survive to an otherwise lethal increase of temperature. A related response is also launched under an abrupt and lethal heat stress that, in this case, is unable to successfully promote thermotolerance and therefore ends up in plant death. Although these molecular programmes are expected to have common players, the overlapping degree and the specific regulators of each process are currently unknown. We have carried out a high-throughput comparative proteomics analysis during acclimation and during the early stages of the plant response to a severe heat stress that lead Arabidopsis seedlings either to survival or death. This analysis dissects these responses, unravels the common players and identifies the specific proteins associated with these different fates. Thermotolerance assays of mutants in genes with an uncharacterized role in heat stress demonstrate the relevance of this study to uncover both positive and negative heat regulators and pinpoint a pivotal role of JR1 and BAG6 in heat tolerance.

Hydrogels provide effective alternatives for drug delivery when therapeutics cannot be applied directly to a wound, or if adverse effects are associated with systemic administration. However, drug delivery vehicles need to be biocompatible and biodegradable and exhibit sufficient mechanical strength to withstand handling and different physiological conditions, such as those encountered during topical administration of a therapeutic. Wound healing can be divided into three phases stimulated by transforming growth factor-beta (TGF-&beta, ) and, subsequently, targeted therapeutics have been developed to inhibit this cytokine for the treatment of chronic wounds and to prevent scarring. In this study, the capacity of calcium alginate hydrogels plasticized with poly(ethylene glycol) methyl ether methacrylate (PEGMA) to deliver anti-TGF-&beta, antibodies (1D11.16.8) to a wound was investigated in situ. Three levels of antibodies, 10, 50, and 100 &mu, g, were loaded into calcium-alginate-PEGMA hydrogels and evaluated in an excisional wound model in mice. Hydrogels containing 50 and 100 &mu, g 1D11.16.8 produced less inflammation, accompanied by a marked reduction in collagen deposition and cell infiltration. These findings demonstrate the capacity of calcium-alginate-PEGMA hydrogels to deliver larger proteins, such as antibodies, to the site of a wound.

Summary Virus infections in plants cause changes in host gene expression that are common to other environmental stresses. In this work, we found extensive overlap in the transcriptional responses between Arabidopsis thaliana plants infected with Tobacco rattle virus (TRV) and plants undergoing senescence. This is exemplified by the up-regulation during infection of several senescence-associated Dark-inducible (DIN) genes, including AtDIN1 (Senescence 1, SEN1), AtDIN6 (Asparagine synthetase 1, AtASN1) and AtDIN11. DIN1, DIN6 and DIN11 homologues were also activated in Nicotiana benthamiana in response to TRV and Potato virus X (PVX) infection. Reduced TRV levels in RNA interference (RNAi) lines targeting AtDIN11 indicate that DIN11 is an important modulator of susceptibility to TRV in Arabidopsis. Furthermore, low accumulation of TRV in Arabidopsis protoplasts from RNAi lines suggests that AtDIN11 supports virus multiplication in this species. The effect of DIN6 on virus accumulation was negligible in Arabidopsis, perhaps as a result of gene or functional redundancy. However, TRV-induced silencing of NbASN, the DIN6 homologue in N. benthamiana, compromises TRV and PVX accumulation in systemically infected leaves. Interestingly, NbASN inactivation correlates with the appearance of morphological defects in infected leaves. We found that DIN6 and DIN11 regulate virus multiplication in a step prior to the activation of plant defence responses. We hypothesize on the possible roles of DIN6 and DIN11 during virus infection.

Aim To investigate the influence of the use L-PRF as a socket filling material and its ridge preservation properties. Materials and Methods Twenty-two patients in need of single bilateral and closely symmetrical tooth extractions in the maxilla or mandible were included in a split-mouth RCT. Treatments were randomly assigned (L-PRF socket filling versus natural healing). CBCT scans were obtained after tooth extraction and three months. Scans were evaluated by superimposition using the original DICOM data. Mean ridge width differences between timepoints were measured at three levels below the crest on both the buccal and lingual sides (crest −1 mm (primary outcome variable), −3 mm and −5 mm). Results Mean vertical height changes at the buccal were −1.5 mm (±1.3) for control sites and 0.5 mm (±2.3) for test sites (p

Objectives This clinical trial explored the clinical and radiographic dimensional changes of the alveolar ridge in the first 4 months after tooth extraction in combination with the application of Leukocyte- and Platelet Rich Fibrin (L-PRF). Methods Eighteen single rooted maxillary and mandibular sockets were filled with L-PRF without soft tissue closure. Clinical measurements (bone sounding) were performed using a customized acrylic stent and radiographic measurements were accomplished using Cone Beam Computed Tomography (CBCT), immediately after tooth extraction and after 4 months. Results The clinical observations indicated a mean horizontal resorption of 1.18 ± 2.4 mm (p = 0.8) at the crest, 1.25 ± 2.0 mm (p = 0.57) and 0.83 ± 2.0 mm (p = 0.78) at 2 mm and 4 mm apical to the crest, respectively. The buccal plate demonstrated a mean vertical loss of 0.44 ± 3.5 mm (p = 0.9), the centre of the socket had a significant filling of 5.72 ± 3.6 mm (p = 0.0001) and the oral cortical plate had a mean vertical gain of 0.09 mm ± 1.57 mm (p = 0.9). The radiographic analysis demonstrated a mean vertical bone loss of 0.27 ± 2.5 mm (p = 0.9) on the buccal and of 0.03 ± 1.6 mm (p = 0.9) at the oral crest. The width of the alveolar ridge had a mean loss of 1.33 mm ± 1.43 mm. Conclusions Within the limitations of this pilot study, it can be concluded that L-PRF might show clinical benefits for ridge preservation.

El sedentarismo es en la actualidad una de las principales fuentes de amenaza para la salud de las personas como efecto de la inactividad física provocando los problemas cardiovasculares, la prevención, tratamiento, observación y recuperación a causa de las enfermedades crónicas no transmisibles; es así que los especialistas en ciencias médicas y del deporte y consecuentemente las autoridades sanitarias recomiendan que toda persona con o sin problemas cardiovasculares incluyan en el desempeño de su vida cotidiana, tanto en el trabajo como en el hogar desarrollen actividad física de forma regular, ya sea para recuperarse o mantener su salud. Por lo tanto, es de considerar los beneficios de la actividad física como herramienta clave para solucionar los problemas propios del sedentarismo causando los problemas cardiovasculares, las nuevas recomendaciones de actividad física para la salud establecen con la aprobación de la OMS, destacan que al menos 30 minutos diarios, por lo menos cinco días de la semana, de forma continua o en sesiones acumulativas de 10 o 15 minutos de una actividad de intensidad moderada.

[EN] Plants have developed versatile strategies to deal with the great variety of challenging conditions they are exposed to. Among them, the regulation of translation is a common target to finely modulate gene expression both under biotic and abiotic stress situations. Upon environmental challenges, translation is regulated to reduce the consumption of energy and to selectively synthesize proteins involved in the proper establishment of the tolerance response. In the case of viral infections, the situation is more complex, as viruses have evolved unconventional mechanisms to regulate translation in order to ensure the production of the viral encoded proteins using the plant machinery. Although the final purpose is different, in some cases, both plants and viruses share common mechanisms to modulate translation. In others, the mechanisms leading to the control of translation are viral- or stress-specific. In this paper, we review the different mechanisms involved in the regulation of translation initiation under virus infection and under environmental stress in plants. In addition, we describe the main features within the viral RNAs and the cellular mRNAs that promote their selective translation in plants undergoing biotic and abiotic stress situations., This work was supported by the ERC Starting Grant 260468 to M. Mar Castellano.

Multifunctional materials and devices found in nature serve as inspiration for advanced synthetic materials, structures and robotics. Here, we elucidate the architecture and unusual deformation mechanisms of seahorse tails that provide prehension as well as protection against predators. The seahorse tail is composed of subdermal bony plates arranged in articulating ring-like segments that overlap for controlled ventral bending and twisting. The bony plates are highly deformable materials designed to slide past one another and buckle when compressed. This complex plate and segment motion, along with the unique hardness distribution and structural hierarchy of each plate, provide seahorses with joint flexibility while shielding them against impact and crushing. Mimicking seahorse armor may lead to novel bio-inspired technologies, such as flexible armor, fracture-resistant structures or prehensile robotics.

The mechanical properties of fully demineralized, fully deproteinized and untreated cortical bovine femur bone were investigated by compression testing in three anatomical directions (longitudinal, radial and transverse). The weighted sum of the stress–strain curves of the treated bones was far lower than that of the untreated bone, indicating a strong molecular and/or mechanical interaction between the collagen matrix and the mineral phase. Demineralization and deproteinization of the bone demonstrated that contiguous, stand-alone structures result, showing that bone can be considered an interpenetrating composite material. Structural features of the samples from all groups were studied by optical and scanning electron microscopy. Anisotropic mechanical properties were observed: the radial direction was found to be the strongest for untreated bone, while the longitudinal one was found to be the strongest for deproteinized and demineralized bones. A possible explanation for this phenomenon is the difference in bone microstructure in the radial and longitudinal directions.

Heat stress is one of the most prominent and deleterious environmental threats affecting plant growth and development. Upon high temperatures, plants launch specialized gene expression programs that promote stress protection and survival. These programs involve global and specific changes at the transcriptional and translational levels. However, the coordination of these processes and their specific role in the establishment of the heat stress response is not fully elucidated. We have carried out a genome-wide analysis to monitor the changes in the translation efficiency of individual mRNAs of Arabidopsis thaliana seedlings after the exposure to a heat shock stress. Our results demonstrate that translation exerts a wide but dual regulation of gene expression. For the majority of mRNAs, translation is severely repressed, causing a decreased of 50% in the association of the bulk of mRNAs to polysomes. However, some relevant mRNAs involved in different aspects of homeostasis maintenance follow a differential pattern of translation. Sequence analyses of the differentially translated mRNAs unravels that some features, such as the 5'UTR G+C content and the cDNA length, may take part in the discrimination mechanisms for mRNA polysome loading. Among the differentially translated genes, master regulators of the stress response stand out, highlighting the main role of translation in the early establishment of the physiological response of plants to elevated temperatures.