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Purpose: The tensiomyography (TMG) technique is increasingly used to determine muscle contractile properties in exercise and injury management. The present study investigated the informative value of TMG parameters in correlation with commonly used (creatine kinase, CK; myoglobin, Mb) and novel candidate biomarkers of muscle damage (heart-type fatty acid-binding protein, h-FABP; high-mobility group box 1, HMGB1)., Methods: Ten untrained men performed 6 × 10 eccentric contractions of the elbow flexors at 110% of the concentric one repetition maximum. CK, Mb, h-FABP, HMGB1, arm circumference, pain and TMG data, including maximal displacement (D m ) and temporal outcomes as the contraction time (T c ), sustained time (T s ), delay time (T d ), and relaxation time (T r ), were assessed pre-exercise, post-exercise, 20 min, 2 h and on the consecutive 3 days post-exercise., Results: CK and h-FABP significantly increased beginning at 24 h, Mb already increased at 2 h (p < 0.05). HMGB1 was only increased immediately post-exercise (p = 0.02). T c and T d remained unchanged, whereas T s and T r were significantly slower beginning at 24 h (p < 0.05). D m was decreased within the first 24 h and after 72 h (p < 0.01). The % change from pre-exercise correlated for D m with CK, Mb, and h-FABP the highest at 48 h (r = - 0.95, - 0.87 and - 0.79; p < 0.01) and for h-FABP with CK and Mb the highest at 24 h (r = 0.96 and 0.94, for all p < 0.001)., Conclusion: This study supports the correlation of TMG parameters with muscle damage markers after eccentric exercise. Therefore, TMG could represent a non-invasive and cost effective alternative to quantify the degree of muscle damage after exercise interventions.

The lubrication effect of bicontinuous microemulsions describes a facilitated flow along hydrophilic planar surfaces because the fluid forms lamellar domains that can slide off along each other much better than randomly ordered domains. The applicability of this effect is based on the prerequisite of the lamellar structure not being destroyed by external shear-fields. In this paper, we demonstrate that the lamellar structure is highly stable for shear rates of up to 600 s -1 using neutron reflectivity and grazing incidence small angle neutron scattering experiments. The transition structure that consists of perforated lamellae is attacked by the flow, and the bicontinuous microemulsion comes closer to the solid interface when shear is applied. All of this verifies the stability of the lubrication effect for technical applications., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

Introduction: In this study we aimed to identify nerve entry points (NEPs) of superficial skeletal muscles obtained by dissection of 20 human cadavers and compared them with motor points (MP) obtained previously by electrical stimulation., Methods: The biceps brachii (BB), trapezius (TZ), latissimus dorsi (LD), pectoralis major (Pmaj), and pectoralis minor (Pmin) muscles were dissected from human cadavers. NEP data (mean ± standard deviation) from each muscle were calculated. F-tests with Bonferroni corrections were used to compare NEPs and MPs., Results: The number of NEPs was 2 in BB, 1 in Pmin, 4 in TZ, and 3 in LD, whereas the total number in Pmaj varied from 3 to 5. NEPs and MPs were statistically equal only in Pmin and in the descending part of TZ., Discussion: The findings show crucial differences between NEPs and MPs, possibly impacting the effectiveness of several medical treatment strategies. Muscle Nerve 57: 460-465, 2018., (© 2017 Wiley Periodicals, Inc.)

Osteoarthritis of the knee is one of the most commonly diagnosed joint ailments and responsible for increased rates of total knee arthroplasty surgeries worldwide. Whereas the surgical approach is able to diminish the perceived knee pain of concerned patients', the postoperative recovery is often accompanied by persistent skeletal muscle dysfunctions and atrophy, which is responsible for functional deficits for up to several years. Recent findings indicate that surgery induced adverse effects on skeletal muscles are largely associated with the use of pneumatic tourniquets, wherefore several studies try to reduce tourniquet use in orthopedic surgery. However, due to comparable incidence of muscle impairment and increased surgical challenge, the most frequently applied surgical technique in TKA is still associated with the use of tourniquets. When attenuating TKA induced adverse effects, the preoperative preparation of patients by specific exercises (called prehabilitation) was able to enhance preoperative overall fitness through associated accelerated recovery. Based on patients' limited functional activity, prehabilitation techniques have to be particularly designed to allow regular adherence. The present paper is based on a narrative review of current literature, and provides a novel hypothesis by which blood flow restriction exercises (BFR) are able to improve patients' compliance to prehabilitation. BFR training is characterized by the application of low-resistance exercise with similar intensities as daily living tasks in association with a suppression of venous blood flow in an extremity, achieving significant morphological and neuromuscular adaptations in skeletal muscles. In addition, preoperative enhancements in muscle health with corresponding benefits in overall fitness, BFR induced molecular alterations could also be able to interfere with TKA induced pathological signaling. Therefore, based on the known major impact of BFR on skeletal muscle physiology, the present paper aims to illustrate the potential beneficial impact of BFR training as a prehabilitation concept to promote patients regular adherence to preoperative exercises and thus achieve an accelerated recovery and increases in patients' satisfaction., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Purpose: Ischemic preconditioning (IPC) is known to reduce muscle damage induced by ischemia and reperfusion injury during surgery. Because of similarities between the pathophysiological formation of ischemia and reperfusion injury and eccentric exercise-induced muscle damage (EIMD), as characterized by an intracellular accumulation of Ca, an increased production of reactive oxygen species, and increased proinflammatory signaling, the purpose of the present study was to investigate whether IPC performed before eccentric exercise may also protect against EIMD., Methods: Nineteen healthy men were matched to an eccentric-only (ECC; n = 9) or eccentric proceeded by IPC group (IPC + ECC; n = 10). The exercise protocol consisted of bilateral biceps curls (3 × 10 repetitions at 80% of the concentric one-repetition maximum). In IPC + ECC, IPC was applied bilaterally at the upper arms by a tourniquet (200 mm Hg) immediately before the exercise (3 × 5 min of occlusion, separated by 5 min of reperfusion). Creatine kinase (CK), arm circumference, subjective pain (visual analog scale score), and radial displacement (tensiomyography, maximal radial displacement) were assessed before IPC, preexercise, postexercise, and 20 min, 2 h, 24 h, 48 h, and 72 h postexercise., Results: CK differed from baseline only in ECC at 48 h (P < 0.001) and 72 h (P < 0.001) postexercise. After 24, 48, and 72 h, CK was increased in ECC compared with IPC + ECC (between groups: 24 h, P = 0.004; 48 h, P < 0.001; 72 h, P < 0.001). The visual analog scale score was significantly higher in ECC at 24-72 h postexercise when compared with IPC + ECC (between groups: all P values < 0.001). The maximal radial displacement was decreased on all postexercise days in ECC (all P values < 0.001) but remained statistically unchanged in IPC + ECC (between groups: P < 0.01)., Conclusions: These findings indicate that IPC performed before a bout of eccentric exercise of the elbow flexors blunts EIMD and exercise-induced pain while maintaining the contractile properties of the muscle.

Purpose: Advantages of tourniquet use in TKA include benefits for surgeons and patients, varying from a bloodless operation site to a reduced intervention time. The time under ischemia and the reperfusion period are crucial phases for affected soft-tissue, most commonly the extensor mechanism., Case Reports: documented its impact on soft-tissue, ranging from necrotic muscle damage to systemic inflammation. Recently, research regarding tourniquet application patterns discuss clinical outcome parameters in the context of soft-tissue damage, excluding the underlying pathophysiological mechanisms., Methods: This review summarizes the molecular aspects of soft-tissue damage occurring during tourniquet application in TKA with special focus on ischemia/reperfusion injury. Recent meta-analyses and original trials were reviewed for data on muscle damage and are presented., Conclusion: Although underlying pathomechanisms are well known and presented, clinical orthopedic research has so far not addressed this issue. In context of physical training, positive effects regarding postoperative recovery might be possible if more attention is paid to prepare involved muscle preoperatively to TKA (prehabilitation).

Eccentric exercise training is effective for increasing muscle mass and strength, and improving insulin sensitivity and blood lipid profiles. However, potential muscle damage symptoms such as prolonged loss of muscle function and delayed onset of muscle soreness may restrict the use of eccentric exercise, especially in clinical populations. Therefore, strategies to reduce eccentric exercise-induced muscle damage (EIMD) are necessary, and an extensive number of scientific studies have tried to identify potential intervention modalities to perform eccentric exercises without adverse effects. The present paper is based on a narrative review of current literature, and provides a novel hypothesis by which an ischemic preconditioning (IPC) of the extremities may reduce EIMD. IPC consists of an intermittent application of short-time non-lethal ischemia to an extremity (e.g. using a tourniquet) followed by reperfusion and was discovered in clinical settings in an attempt to minimize inflammatory responses induced by ischemia and ischemia-reperfusion-injury (I/R-Injury) during surgery. The present hypothesis is based on morphological and biochemical similarities in the pathophysiology of skeletal muscle damage during clinical surgery and EIMD. Even though the primary origin of stress differs between I/R-Injury and EIMD, subsequent cellular alterations characterized by an intracellular accumulation of Ca 2+ , an increased production of reactive oxygen species or increased apoptotic signaling are essential elements for both. Moreover, the incipient immune response appears to be similar in I/R-Injury and EIMD, which is indicated by an infiltration of leukocytes into the damaged soft-tissue. Thus far, IPC is considered as a potential intervention strategy in the area of cardiovascular or orthopedic surgery and provides significant impact on soft-tissue protection and downregulation of undesired excessive inflammation induced by I/R-Injury. Based on the known major impact of IPC on skeletal muscle physiology and immunology, the present paper aims to illustrate the potential protective effects of IPC on EIMD by discussing possible underlying mechanisms., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

In sports medicine, muscle enzymes in the blood are frequently used as an indicator of muscle damage. It is commonly assumed that mechanical stress disrupts plasma membrane to an extent that allows large molecules, such as enzymes, to leak into the extracellular space. However, this does not appear to fully explain changes in muscle enzyme activity in the blood after exercise. Apart from this mechanically induced membrane damage, we hypothesize that, under critical metabolic conditions, ATP consuming enzymes like creatine kinase (CK) are "volitionally" expulsed by muscle cells in order to prevent cell death. This would put themselves into a situation comparable to that of CK deficient muscle fibers, which have been shown in animal experiments to be virtually infatigable at the expense of muscle strength. Additionally we expand on this hypothesis with the idea that membrane blebbing is a way for the muscle fibers to store CK in fringe areas of the muscle fiber or to expulse CK from the cytosol by detaching the blebs from the plasma membrane. The blebbing has been shown to occur in heart muscle cells under ischaemic conditions and has been speculated to be an alternative pathway for the expulsion of troponin. The blebbing has also been seen skeletal muscle cells when intracellular calcium concentration increases. Cytoskeletal damage, induced by reactive oxygen species (ROS) or by calcium activated proteases in concert with increasing intracellular pressure, seems to provoke this type of membrane reaction. If these hypotheses are confirmed by future investigations, our current understanding of CK as a blood muscle damage marker will be fundamentally affected., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Objectives: In the revised version of ISO 7405 there are so far no detailed recommendations concerning temperature and humidity during specimen production for light curing and chemically setting dental materials. The main objective of the present study was to observe if different environmental conditions during specimen production influence cytotoxicity and degree of conversion of four post and core composite materials and to investigate if cytotoxicity of post and core materials is influenced by their corresponding bonding substances., Methods: Specimens of four different post and core composite materials (LuxaCore - Dual, Core X-Flow, Flow White and MultiCore Flow) were produced in a climate test chamber at 23°C/50% relative humidity or 37°C/95% relative humidity and were dual-cured or self-cured, with or without their corresponding bonding substances. Specimens were added to cell cultures immediately after production or after preincubation for 7 days. Specimens were incubated with L-929 fibroblasts for 72h and cell numbers determined by a flow cytometer. FTIR spectroscopic measurements of post and core materials were performed at the same temperature conditions as for the cytotoxicity assay (23°C or 37°C)., Results: Dual-cured specimens of all post and core composites exhibited less cytotoxicity under both environmental conditions than self-cured specimens. All self-cured specimens manufactured at 37°C/95% showed less cytotoxicity than specimens produced at 23°C/50%. All dual-cured specimens showed similar cytotoxicity at both environmental conditions. After 7 days of preincubation most dual-cured specimens produced at 23°C/50% showed less cytotoxicity than self-cured specimens (with the exception of Flow White). Compared to fresh specimens, 7-day aged specimens of most materials showed reduced cytotoxicity. Materials already showing low cytotoxicity as fresh specimens did not further reduce their cytotoxicity after 7 days of preincubation. For dual-cured materials the degree of conversion was higher compared to self-cured materials., Significance: Different temperatures during specimen production have an impact on cytotoxicity and degree of conversion of dual-curing composite materials. Detailed recommendations for standardization concerning environmental conditions during specimen production are required., (Copyright © 2014 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.)

In 2006, we reported a mariner (Mos1)-transformed Aedes aegypti line, Carb77, which was highly resistant to dengue-2 virus (DENV2). Carb77 mosquitoes expressed a DENV2-specific inverted-repeat (IR) RNA in midgut epithelial cells after ingesting an infectious bloodmeal. The IR-RNA formed double-stranded DENV2-derived RNA, initiating an intracellular antiviral RNA interference (RNAi) response. However, Carb77 mosquitoes stopped expressing the IR-RNA after 17 generations in culture and lost their DENV2-refractory phenotype. In the current study, we generated new transgenic lines having the identical transgene as Carb77. One of these lines, Carb109M, has been genetically stable and refractory to DENV2 for >33 generations. Southern blot analysis identified two transgene integration sites in Carb109M. Northern blot analysis detected abundant, transient expression of the IR-RNA 24 h after a bloodmeal. Carb109M mosquitoes were refractory to different DENV2 genotypes but not to other DENV serotypes. To further test fitness and stability, we introgressed the Carb109M transgene into a genetically diverse laboratory strain (GDLS) by backcrossing for five generations and selecting individuals expressing the transgene's EGFP marker in each generation. Comparison of transgene stability in replicate backcross 5 (BC5) lines versus BC1 control lines demonstrated that backcrossing dramatically increased transgene stability. We subjected six BC5 lines to five generations of selection based on EGFP marker expression to increase the frequency of the transgene prior to final family selection. Comparison of the observed transgene frequencies in the six replicate lines relative to expectations from Fisher's selection model demonstrated lingering fitness costs associated with either the transgene or linked deleterious genes. Although minimal fitness loss (relative to GDLS) was manifest in the final family selection stage, we were able to select homozygotes for the transgene in one family, Carb109M/GDLS.BC5.HZ. This family has been genetically stable and DENV2 refractory for multiple generations. Carb109M/GDLS.BC5.HZ represents an important line for testing proof-of-principle vector population replacement.

Aedes aegypti is the principal vector of dengue virus (DENV) throughout the tropical world. This anthropophilic mosquito species needs to be persistently infected with DENV before it can transmit the virus through its saliva to a new vertebrate host. In the mosquito, DENV is confronted with several innate immune pathways, among which RNA interference is considered the most important. The Ae. aegypti genome project opened the doors for advanced molecular studies on pathogen-vector interactions including genetic manipulation of the vector for basic research and vector control purposes. Thus, Ae. aegypti has become the primary model for studying vector competence for arboviruses at the molecular level. Here, we present recent findings regarding DENV-mosquito interactions, emphasizing how innate immune responses modulate DENV infections in Ae. aegypti . We also describe the latest advancements in genetic manipulation of Ae. aegypti and discuss how this technology can be used to investigate vector transmission of DENV at the molecular level and to control transmission of the virus in the field.

Transgenic mosquitoes generated by transposable elements (TEs) often poorly express transgenes owing to position effects. To avoid these effects, the ΦC31 site-directed recombination system was used to insert transgenes into a locus favourable for gene expression in Aedes aegypti. We describe phenotypes of mariner Mos1 TE and ΦC31 transgenic mosquitoes expressing the enhanced green fluorescent protein (EGFP) reporter in midguts of blood-fed females. Mosquitoes of nine TE-generated lines [estimated transformation frequency (TF): 9.3%] clearly expressed the eye-specific selection marker but only 2/9 lines robustly expressed the EGFP reporter. The piggyBac TE-generated ΦC31 docking strain, attP26, supported recombination with attB site containing donors at an estimated TF of 1.7-4.9%. Using a codon-optimized ΦC31 integrase mutant instead of the 'wild-type' enzyme did not affect TF. Site-directed recombination of line attP26 with an attB-containing donor expressing EGFP from the Ae. aegypti carboxypeptidase promoter produced one transgenic line with blood-fed females expressing the reporter in midgut tissue. Docking strain attP26 also supported robust expression of Flock House virus B2 from the Ae. aegypti polyubiquitin promoter. Our data confirm that eye-specific selection marker expression alone is not a reliable indicator for robust gene-of-interest expression in Ae. aegypti and that the ΦC31 system can ensure predictable transgene expression in this mosquito species., (© 2011 The Authors. Insect Molecular Biology © 2011 The Royal Entomological Society.)

Objectives: Assessment of dental material biocompatibility is gaining increasing importance for both patients and dentists. Dental cements may be in contact with oral soft tissues for prolonged periods of time and play an important role in prosthetic rehabilitation. The aim of the present study was to evaluate eight dental cements using a standardized L929-fibroblast cell culture test., Methods: For each material, fresh specimens (added to the cultures immediately after preparation) and specimens preincubated for 7 days in cell culture medium were prepared according to the manufacturers' recommendations. After exposure to test specimens, cell numbers were compared to glass controls. The main outcome was a two-sided 95% confidence interval for the mean value of the standardized cell number for each substance investigated., Results: Fresh specimens of all tested cements showed significant cytotoxicity, which diminished after 7 days preincubation. Cytotoxicity of fresh adhesive and self-adhesive resin cements was lower when specimens were dual-cured compared to self-cured. A rank order of cytotoxicity was established based on mean values: Nexus 2 (dual-cured) showed least cytotoxicity, followed by Variolink II (dual-cured), Nexus 2 (self-cured), Harvard, RelyxUnicem (dual-cured), Panavia 21, Fujicem, Durelon, Variolink II (self-cured), RelyxUnicem (self-cured), Maxcem (dual-cured) and Maxcem (self-cured). When bondings were added to Nexus 2 or Variolink II specimens, a slight increase in cytotoxicity was observed., Significance: Adhesive resin cements showed less cytotoxicity than self-adhesive and chemically setting cements. Bonding only slightly influenced cytotoxicity of the adhesive resin cements. Dual-cured specimens of adhesive and self-adhesive resin cements showed significantly less toxicity than self-cured specimens.

Objectives: Recently, we found that dental bonding substances tested alone or in combination with composites are far more cytotoxic than composite materials alone. These data are in line with several cytotoxicity reports but contradict in vivo studies showing the beneficial effects of bonding materials. The aims of the present study were to develop a preparation method for composite specimens modelling conditions in the oral cavity and to analyse the influence of bonding substances on the cytotoxicity of six different composite materials., Methods: Cylindrical composite specimens were prepared in polyethylene blocks containing 5 mm diameter cylindrical holes (cylinder height 2 mm), covered with a polyethylene foil and light cured from one end for 40 s. In a second series of experiments, composite specimens were combined with bonding materials. Bonding was applied onto the polyethylene foil in one or two layers and light cured according to the manufacturers' instructions. Subsequently, polyethylene moulds were placed on top of the bonding materials and composites prepared as described above. After unilateral light curing from the top of the cylindrical holes, visual confirmation of adherence at the base was obtained. Specimens were added to the cultures immediately after production or after preincubation for 7 days under cell culture conditions. Specimens were incubated with L-929 fibroblasts for 72 h and cell numbers determined by flow cytometry. To evaluate the degree of conversion (DC) of bonding materials cured with and without air inhibition a third series of experiments was performed. FTIR spectroscopic measurements were made on thin-films of dentin-bonding agents, cured under both an-aerobic and aerobic conditions, to determine degree of conversion., Results: Cytotoxicities of all six tested composites were significantly different (p<0.0001) and diminished after 7 days of preincubation (p<0.0001). Bonding substances had no statistically significant influence on the cytotoxicity of composite materials (p=0.159). A highly significant statistical reduction in the degree of conversion for each resin cured under air inhibition conditions was documented (p<0.01)., Significance: Our study demonstrates that cell culture toxicity data are highly model dependent and that internationally standardized test protocols for toxicity screening of dental materials in line with the existing standards are clearly needed to obtain comparable results.

The objective of this study was to compare the cytotoxicity of four orthodontic bonding systems, Light Bond, Enlight, Concise, and Transbond, and to evaluate their shear bond strength (SBS). These orthodontic bonding materials were applied to metal brackets (Mini Diamond). Glass specimens were used as controls in all experiments. Only Concise was a chemically cured system, the other systems were light cured. The specimens were added to L-929 fibroblast cultures immediately after fabrication or after pre-incubation for 7 days. The incubation time was 72 hours and the cells were counted by flow cytometry. One hundred and fifty-seven freshly extracted human third molars were used for testing the SBS in a universal testing machine. Statistical significance was determined using analysis of variance followed by post hoc comparisons for multiple-level alpha control. Pairwise comparisons showed a significant difference only between Light Bond and Concise (P = 0.0126). The highest SBS was obtained with Light Bond (23.23 +/- 1.53 MPa) followed by Transbond (20.39 +/- 1.18 MPa) and Enlight, (20.32 +/- 1.06 MPa). Concise (17.87 +/- 1.04 MPa) showed the lowest SBS. The cytotoxicity of all light-cured systems for fresh specimens was comparable, whereas the chemically cured system, Concise, was significantly more cytotoxic. After 7 days of pre-incubation, all systems were significantly less cyotoxic than fresh specimens (P < 0.001). Brackets alone were not cytotoxic. All bonding systems showed a clinically satisfactory bond strength higher than 10 MPa, with the chemically cured system showing the lowest SBS.

Objectives: The standardization protocols for biomaterial cytotoxicity testing require fine tuning for oral biomaterials to obtain international comparability as the basis for risk assessment. The principal aims were specifically to evaluate the effect of (i) relative interface area (ratio of specimen surface to cell layer surface) and (ii) volume of cell culture medium on cytotoxicity as a potential modification of ISO 10993-5., Methods: ISO 10993-5 was followed with an interface area of 12.5%, as recommended, using primary human gingival fibroblasts and L-929 mouse fibroblasts. In another series of experiments (using L-929 cells) the interface area was varied between 12.5% and 0.71%. For each relative interface area, three conditions for affecting the cure of the resin composite were investigated by using three mould materials: white, transparent and black moulds. In addition, the volume of cell culture medium was varied. Composite specimens (Herculite XRV) were added to the cultures immediately after production or preincubation for 1, 2, 7 days or 6 weeks under cell culture conditions. Specimens were incubated with fibroblasts for 72 h and cell numbers determined by flow cytometry. Glass specimens resembling composite specimens in diameter and height were used as negative controls., Results: Cytotoxicity results with primary gingival fibroblasts were comparable to results with the cell line L-929. An effect from the color/material of the specimen moulds was found. Different ratios of specimen sizes to cell culture parameters (cell layer surface, volume of cell culture medium) produced different results. Three out of four differently designed specimens showed the same behavior in cell culture., Significance: Cytotoxicity tests should be further standardized in line with existing standards with regard to specimen production protocols to ensure results are internationally comparable to validate these tests as tools for risk assessment.

Mosquitoes (Aedes aegypti) were genetically modified to exhibit impaired vector competence for dengue type 2 viruses (DENV-2). We exploited the natural antiviral RNA interference (RNAi) pathway in the mosquito midgut by constructing an effector gene that expresses an inverted-repeat (IR) RNA derived from the premembrane protein coding region of the DENV-2 RNA genome. The A. aegypti carboxypeptidase A promoter was used to express the IR RNA in midgut epithelial cells after ingestion of a bloodmeal. The promoter and effector gene were inserted into the genome of a white-eye Puerto Rico Rexville D (Higgs' white eye) strain by using the nonautonomous mariner MosI transformation system. A transgenic family, Carb77, expressed IR RNA in the midgut after a bloodmeal. Carb77 mosquitoes ingesting an artificial bloodmeal containing DENV-2 exhibited marked reduction of viral envelope antigen in midguts and salivary glands after infection. DENV-2 titration of individual mosquitoes showed that most Carb77 mosquitoes poorly supported virus replication. Transmission in vitro of virus from the Carb77 line was significantly diminished when compared to control mosquitoes. The presence of DENV-2-derived siRNAs in RNA extracts from midguts of Carb77 and the loss of the resistance phenotype when the RNAi pathway was interrupted proved that DENV-2 resistance was caused by a RNAi response. Engineering of transgenic A. aegypti that show a high level of resistance against DENV-2 provides a powerful tool for developing population replacement strategies to control transmission of dengue viruses.

Objective: Advanced dental packable and nonpackable composite materials have been recently introduced onto the market as 'amalgam alternatives' for the restoration of posterior teeth. Most established composites are applied in increments of no more than 2mm and light cured to ensure complete polymerization, whereas new formulations have been claimed to be suitable for application in increments of 4-5mm. The aims of the present study were to analyze the cytotoxicity of these new composites in comparison to an established nonpackable composite using standardized cell culture systems and to examine the influence of thickness of the light cured increments on the cytotoxicity of these materials., Methods: Specimens were prepared in polyethylene tubes covered with mylar. All materials were light cured in 2.5 or 5mm increments with a Demetron curing light (light intensity: 550 mW/cm(2)). Specimens were added to the cultures immediately after production or after preincubation for 1, 2, 7 days or 6 weeks under cell-culture conditions. Specimens were incubated with L-929 fibroblasts for 72 h and cell numbers determined by flow cytometry. In a different series of experiments, dopaminergic cells were incubated with composite supernatants., Results: Results with L-929 fibroblasts demonstrated that all freshly prepared composite materials reduced cell numbers (p<0.05) in comparison to controls. The cytotoxicity of all substances diminished after increasing preincubation times (p<0.0001). In a rank order of cytotoxicity, the established composite and two of the advanced composite materials exhibited least cytotoxicity, whereas the other advanced composites showed moderate or severe cytotoxic effects. The cytotoxicity of each material was much higher when polymerized in 5mm increments than in 2.5mm composite increments (p<0.0001)., Significance: Our data demonstrate that the advanced composites tested show similar or more severe cytotoxicity than an established nonpackable composite and that cytotoxicity of all materials investigated increases when applied in a 5mm bulk increment.