Holm-Sidak Test Research Articles

Mucoadhesive Polymers Enhance Ocular Drug Delivery: Proof of Concept Study with 0.5% Tropicamide in Dogs.

Purpose: To assess the efficacy of 0.5% tropicamide applied after topical administration of 1.4% hydroxyethyl cellulose (HEC) or 1.2% hyaluronic acid (HA) at different time intervals. Methods: Eleven healthy Labrador retriever dogs were used. The study consisted of 7 trials in which pupil diameter (PD) was measured every 30 min for 7 h after drug administration. In trial 1, PD was measured after tropicamide application (control). In trials 2-7, PD was measured after treatment with tropicamide 10 s, 1 and 5 min after application of HEC or HA. Data were analyzed using analysis of variance and post hoc Holm-Sidak tests. Results: Maximal PD (mean ± standard deviation) was significantly greater (P < 0.001) in 5 of 6 experimental trials, with the greatest PD recorded when HEC was applied 1 min (12.2 ± 0.4 mm) and HA 10 s (12.3 ± 0.5 mm) before tropicamide, compared with 11.2 ± 0. 7 mm in control trial 1. PD >10 mm was maintained for 2.75-5 h in trials 2-7 and 2.25 h in trial 1 (P < 0.001). Area under the PD-time curve ranged from 61.9 to 71.3 ± 2.9 mm·hours in trials 2-7, significantly greater than 59.3 ± 1.8 mm·hours in control trial 1 (P ≤ 0.031). Conclusions: Tropicamide efficacy was enhanced by prior administration of a mucoadhesive polymer. Additional studies are needed to determine if HEC and HA of varying concentrations similarly extend and potentiate the effect of other topical ophthalmic drugs in canine eyes.

Lost-wax or laser sintering? Findings on marginal and internal adaptation of metallic copings

Marginal and internal adaptation are parameters of crucial importance to the success of prosthetic crowns. Automatized process creates an expectative of superior or equivalent results compared to restorations manufactured ​​by conventional lost-wax technique. The purpose of this study was to evaluate the marginal adaptation and internal adaptation (cement space) of metal-ceramic crown copings produced by lost-wax (LW) and direct metal laser sintering (DMLS) techniques. An artificial lower first molar was prepared for a full crown, duplicated in plaster and scanned. Twenty metal-ceramic crown copings were fabricated in cobalt-chromium by the two techniques (n=10). The copings were filled with low viscosity silicone and seated on the prepared tooth, resulting in a replica of the internal space. The pellicle formed was embedded in heavy body silicone, sectioned and captured by means of a stereomicroscope at 50x magnification, according to replica technique (RT). Shapiro-Wilk test followed by Holm-Sidak test were used for statistical analysis (α=.05). Marginal adaptation presented no difference between LW (101.5 ± 41.6) and DMLS (86.3 ± 39.9) groups (p=0.24). Conventional LW technique showed significantly lower occlusal (p&lt;0.008) and axial spaces (p&lt;0.03). Measurements of all regions showed numerically larger adaptation values than that defined during design​​ for DMLS group. Both the LW technique and the DMLS technique are within the clinically acceptable.

The Effect of Touch-Cure Polymerization on the Conversion and Hardness of Core Build-Up Resin Composites: A Laboratory Study.

To improve the self-curing capacity and interfacial strength with dentine of dual-cured composite materials, touch-cure activators have been introduced. The aim of the study was to evaluate the effect of these activators on the hardness and conversion of dual-cured resin composite core build-up restoratives. The materials tested were Clearfil DC Core Plus (CF) and Gradia Core (GC) with the corresponding adhesives Clearfil S3 Bond Plus (for CF) and G-Premio Bond/G-Premio DCA activator (for GC). Disk-shaped specimens (n = 6/group) were prepared for the following groups: dual-cured, self-cured and self-cured in contact with the adhesive activators at the bottom surface. After a 3-week storage period (dark/dry/37 °C) the Martens hardness (HM) and degree of conversion (DC%) were determined for the previously mentioned groups and the top surfaces of groups in contact with the adhesives. A statistical analysis was performed by a one-way ANOVA and Holm–Sidak test per material and a Pearson’s correlation analysis (HM vs. DC%) at an α = 0.05. The self-cured specimens resulted in significantly lower HM and DC% values from the dual-cured group, as expected. However, in the presence of the adhesives with touch-cure activators, the conversion of the self-cured groups showed insignificant differences in HM and DC% from the dual-cured in both composite materials. The improvements on the bottom composite surfaces in contact with the adhesives did not extend to the entire specimen length. Nevertheless, improved interfacial curing may improve interfacial durability.

Formation of hydroxyapatite nanoprecursors by the addition of bioactive particles in resin-modified glass ionomer cements

The objective of this study was to evaluate the impact of adding bioactive glasses 45S5 and niobophosphate (NbG) in different concentrations (5, 10 and 20%) of two resin modified glass ionomer cements (RMGIC). Two RMGIC were used: Vitro Fil LC (DFL) and Resiglass F (Biodinâmica) they were modified by the addition of two bioactive glasses - 45S5 or NbG. Different concentrations of bioactive glasses (5, 10 and 20 wt%) were mixed with the powder of the RMGICs. The properties evaluated were: alkalinization activity (pH), fluoride (F−), calcium (Ca2+) and phosphate (PO43−) release, flexural strength, flexural modulus, Knoop microhardness and bioactivity. The analyses of the surfaces were examined using SEM and FTIR after storage at 37 °C for 28 days in simulated body fluid (SBF). The data were analyzed using ANOVA One-way (multiple comparison vs control) and Holm-Sidak test (α = 0.05). The addition of 45S5 increased the pH while the addition of NbG neutralized the pH of the Vitro Fil RMGIC. The RMGIC Resiglass maintained its acidic pH condition in almost all conditions. The addition of the 45S5 and NbG in high concentration increased the release of F−, Ca2+ and PO43− ions. Resiglass did not reduce its mechanical properties with the addition of bioactive glasses (p > 0.05). NbG glass did not alter the mechanical properties of the Vitro Fil (p > 0.05). The addition of the bioactive glasses 45S5 and NbG contributed to the formation of nanoprecursors of hydroxyapatite. The obtaining of bioactive RMGIC is dependent on the composition of the material. The NbG glass provides a neutralization of the pH, an increase of the ions release and the amount of precipitates without jeopardizing the mechanical properties of the RMGIC.

A Multi-Objective Anti-Predatory NIA for E-Commerce Logistics Optimization Problem

Nature-inspired algorithms (NIAs) have established their promising performance to solve both single-objective optimization problems (SOOPs) and multi-objective optimization problems (MOOPs). Anti-predatory NIA (APNIA) is one of the recently introduced single-objective algorithm based on the self-defense behavior of frogs. This paper extends APNIA as multi-objective algorithm and presents the first proposal of APNIA to solve MOOPs. The proposed algorithm is a posteriori version of APNIA, which is named as multi-objective anti-predatory NIA (MO-APNIA). It uses the concept of Pareto dominance to determine the non-dominated solutions. The performance of the MO-APNIA is established through the experimental evaluation and statistically verified using the Friedman rank test and Holm-Sidak test. MO-APNIA is also employed to solve a multi-objective variant of hub location problem (HLP) from the perspective of the e-commerce logistics. Results indicate that the MO-APNIA is also capable to finds the non-dominated solutions of HLP. This finds immense use in logistics industry.

Assessment of the survival and success rates of lithium disilicate crowns after different surface finishing procedures: An in vitro study

Statement of problemEvidence is limited for the impact of clinical adjustments and polishing on the longevity of glazed lithium disilicate restorations. PurposeThe purpose of this in vitro study was to evaluate the influence of surface finishing on the survival and success rates of lithium disilicate restorations based on fatigue resistance and failure mode. Material and methodsLithium disilicate (IPS e.max CAD) maxillary premolar crowns (N=54) were cemented on a dentin analog. The restorations were divided into 3 groups: overglaze (OG), abrasion (GA), and abrasion and polishing (AP). The crowns were submitted to cyclic fatigue in 37 oC water at 100 N and 2 Hz in 2 lifetimes. The load was applied to the occlusal surface by using anatomic pistons to simulate a clinical tripod occlusal contact. After cycling, the crowns were examined for failure (cracking, chipping, or catastrophic fractures) under optical and scanning electron microscopy. Cracking was considered either a structural failure (success analysis) or a survival (clinical criteria - survival analysis). Data were analyzed by using the log rank Kaplan-Meier and Holm-Sidak tests (α=.05). ResultsSurface finishing had no influence on the structural integrity of lithium disilicate, with similar success rates (P=.720). The calculated survival rate was higher for AP than that for other groups (P=.028). Cracking was found for GA and AP crowns, mostly initiating from the external surface. Chipping occurred in all experimental groups, and AP crowns did not show catastrophic failures. ConclusionsAlthough surface treatments had no influence on the success of lithium disilicate, polishing showed a positive effect on the survival rate of the crowns based on the clinical implications of cracking (no need for replacement).

Energy-Dispersive X-Ray Spectrometry Analysis and Radiopacity of Five Different Root Canal Sealers.

Root canal sealers should have optimum adhesion to the root dentin, low cytotoxicity, sufficient radiopacity and high dimensional stability, which depend mainly on their chemical composition. The aims of this study were to evaluate the chemical composition of Syntex (Cerkamed, Stalowa Wola, Poland), Adseal (Meta Biomed, Cheongju-si, Korea), Sealapex (SybronEndo, Scafati, Italy), MTA Fillapex (Angelus, Londrina, Brazil) and GuttaFlow Bioseal (Coltene/Whaledent, Altstätten, Switzerland) and to determine their radiopacity comparatively. Stainless steel molds with an inner diameter of 5 mm and a depth of 2 mm were constructed. Tested root-canal sealers were mixed and poured into the molds. Following setting, digital periapical radiographs of the specimens (n=12) were taken under standard exposure conditions and the mean gray-values of specimens were measured using a computer graphics program. Then the specimens were examined by scanning electron microscopy, and Energy-dispersive X-ray analysis was performed. The data were analyzed using One-way ANOVA followed by Holm-Sidak multi-comparison test (p = 0.05). Sealapex showed higher calcium peak than MTA Fillapex, GuttaFlow Bioseal, Adseal and Syntex sealers. MTA Fillapex contained a highest amount of tungstate. Sealapex presented the highest radiopacity value among the tested materials (p < 0.05), whereas MTA Fillapex was the least radiopaque material (p < 0.05). Each root canal sealer contains different compositions of radiopacifiers including bismuth oxide, calcium tungstate and zirconium oxide. The tested root canal sealers had radiopacity values above the ISO-specified limit. Sealapex was the most radiopaque root canal sealer followed by GuttaFlow Bioseal, Syntex, Adseal and MTA Fillapex.

P–372 Maternal circadian disruption is associated with impaired rhythmic expression of molecular clock genes and IUGR during placenta development in mice

Abstract Study question Are molecular clock genes (MCGs) expressed rhythmically in mouse placenta, and whether maternal circadian rhythm disruption (MCRD) is associated with intrauterine growth retardation (IUGR) through disturbing rhythmic expression of MCGs? Summary answer Maternal circadian disruption causes impaired rhythmic expression of MCGs (Bmal1, Clock, Npas2, Per1, Per2, Per3, Cry1, and Cry2) and IUGR during placenta development in mice. What is known already The world economy is based on a 24/7 society and shift work or jet travel across time zones disrupts circadian rhythm in pregnant women. Evidence indicates that gestational chrono-disruption results in IUGR. Mature mouse and human placenta express MCGs. There is no information in the literature on whether the MCG expression in the placenta is rhythmic or not and whether the rhythmic expression of MCGs is impaired due to MCRD during pregnancy. Also, it is not known whether the association with MCRD and IUGR is related to MCGs. Study design, size, duration Young adult female BALB/c mice were paired with males until vaginal plug formation was verified. Females were randomly assigned to two groups: control and phase-advance. Controls remained on a constant 12-hr light:12-hr dark cycle, whereas phase-advanced mice were subjected to 6-hr advances in the LD cycle every 5 days. Placentae (n = 1329) and fetuses were obtained from 144 mice at Zeitgeber time (ZT)0, ZT6, ZT12, and ZT18 days 12, 14, and 16 of pregnancy. Participants/materials, setting, methods The following analysis was performed: (i) open field test was used for locomotor activity evaluations to confirm MCRD, (ii) placenta/fetus weight ratio for evaluation of IUGR development, (iii) morphometric evaluation of placental compartments utilizing H&amp;E staining (iv) gene expression analysis of MCGs utilizing qRT-PCR. One-way and Two-way ANOVA test followed by Holm-Sidak posthoc test was used for multiple comparisons. Values are expressed as mean ± standard error, and values below p &amp;lt; 0.05 were considered statistically significant. Main results and the role of chance Expression of MCGs (Bmal1, Clock, Npas2, Per1, Per2, Per3, Cry1, and Cry2) was rhythmic in the early and mature placenta development stages (days 12, 14, 16). Locomotor activity tests reveal that the total distance covered on the 16th day of pregnancy significantly decreased compared to the control group (p = 0.000158). The ratio of the time spent in the outer/inner quadrant, an anxiety indicator, significantly increased in the MCRD group on the 14th (p = 0.0351) and 16th days of pregnancy (p = 0.000329). While the number of fetuses was similar in both groups for all gestational days (p = 0.896), in the MCRD group, the fetus/placenta weight ratio decreased significantly on the 12th and 16th days of pregnancy (p &amp;lt; 0.001). Thus, IUGR developed due to MCRD. Histomorphometry analysis of the placental compartments revealed a significant reduction in the spongiotrophoblast layer’s size on all days of pregnancy and the labyrinth layer on day 16 (p &amp;lt; 0.05). Finally, the rhythmic expression MCGs were impaired in placentas obtained from MCRD groups on days 12th, 14th, 6th of pregnancy (p &amp;lt; 0.001). In conclusion, we found a robust relationship with the disturbed MCGs expression and occurrence of IUGR during a chrono-disrupted gestation. Limitations, reasons for caution Since this study was conducted in mice, care should be taken when translating the results to humans. Wider implications of the findings: Our results in mice are important for initiating basic science knowledge regarding the outcomes of maternal chrono-disruption. Moreover, research in the placenta of gestational chrono-disrupted mothers, such as shift-workers, are urgently needed to translate our findings into the clinic. Trial registration number TUBITAK–119S121 and Akdeniz University Research Projects Unit TYL–2018–3960

Brain derived neurotrophic factor mediates accelerated recovery of regenerative electrical stimulation in an animal model of stress urinary incontinence

ObjectiveStress urinary incontinence (SUI) is prevalent among older women and can result from insufficient regeneration of the pudendal nerve (PN). Electrical stimulation (ES) of the PN upregulates brain derived neurotrophic factor (BDNF) and accelerates regeneration. Using tyrosine kinase B (TrkB) to reduce the availability of free BDNF, the aim of this study was to determine if BDNF is necessary for accelerated recovery via ES in a model of SUI. MethodsOur SUI model consists of Female Sprague-Dawley rats, whose PNs were crushed bilaterally twice for 30 s, followed by insertion of a modified Foley catheter into the vagina with balloon inflation for 4 h. These rats were divided into 4 groups: 1) Sham PN crush and sham vaginal distension without electrode implantation and with saline treatment (sham injury); 2) SUI with sham stimulation and saline treatment (SUI); 3) SUI and ES with saline treatment (SUI&ES); and 4) SUI and ES with TrkB treatment (SUI&ES&TrkB). Animals underwent ES or sham stimulation four times a week for two weeks. Four weeks after injury, animals underwent functional testing consisting of leak point pressure (LPP) with simultaneous external urethral sphincter (EUS) electromyography (EMG) and pudendal nerve recordings. Data was analyzed using ANOVA with Holm-Sidak posthoc test (p < 0.05). EUS and PN specimen were sectioned and stained to semi-quantitatively evaluate morphology, regeneration, and reinnervation. ResultsLPP and EUS EMG firing rate were significantly increased in the sham injury and SUI&ES groups compared to the SUI and SUI&ES&TrkB groups. EUS of SUI rats showed few innervated neuromuscular junctions compared to sham injured rats, while both treatment groups showed an increase in reinnervated neuromuscular junctions. ConclusionES accelerates functional recovery via a BDNF-mediated pathway in a model of SUI. These findings suggest ES could be used as a potential regenerative therapy for women with SUI.

Efficacy of a Next Generation Quaternary Ammonium Chloride Sanitizer on Staphylococcus and Pseudomonas Biofilms and Practical Application in a Food Processing Environment

Foodborne pathogens are known to adhere strongly to surfaces and can form biofilms in food processing facilities; therefore, their potential to contaminate manufactured foods underscores the importance of sanitation. The objectives of this study were to (1) examine the efficacy of a new-generation sanitizer (Decon7) on Staphylococcus and Pseudomonas biofilms, (2) identify biofilm bacteria from workers’ boots in relation to previous sanitizer chemistry, (3) validate the efficacy of Decon7 on biofilm from workers’ boots from an abattoir/food processing environment, and (4) compare the sensitivity of isolated boot biofilm bacteria to new- and early (Bi-Quat)-generation QAC sanitizers. Decon7 was applied at two concentrations (5%, 10%) and was shown to be effective within 1 min of exposure against enhanced biofilms of Staphylococcus spp. and Pseudomonas spp. in 96-well microplates. Decon7 was also used to treat workers’ boots that had accumulated high levels of biofilm bacteria due to ineffective sanitization. Bacteria isolated before enzyme/sanitizer treatment were identified through 16S rRNA PCR and DNA sequencing. All treatments were carried out in triplicate and analyzed by one-way RM-ANOVA or ANOVA using the Holm–Sidak test for pairwise multiple comparisons to determine significant differences (p &lt; 0.05). The data show a significant difference between Decon7 sanitizer treatment and untreated control groups. There was a ~4–5 log reduction in Staphylococcus spp. and Pseudomonas spp. (microplate assay) within the first 1 min of treatment and also a &gt; 3-log reduction in the bacterial population observed in the biofilms from workers’ boots. The new next-generation QAC sanitizers are more effective than prior QAC sanitizers, and enzyme pre-treatment can facilitate biofilm sanitizer penetration on food contact surfaces. The rotation of sanitizer chemistries may prevent the selective retention of chemistry-tolerant microorganisms in processing facilities.

Renal Oxygen Sensing Mechanisms May Contribute to Maintaining Cerebral Perfusion During Acute Anemia

Introduction The integrative physiological response to anemia is complex and incompletely understood. We utilized data from studies of acute anemia in rodents to determine the relationship between changes in blood oxygen content (CaO2) and the heterogenous response of the kidney, heart and brain. We hypothesize that renal hypoxia sensing mechanisms contribute to adaptive physiological responses to maintain cerebral oxygen delivery (DO2) during acute anemia. Methods With animal care committee approval, we synthesized novel and published data from 5 previously published studies. Outcomes included: assessment of the relationship between CaO2 and microvascular renal and brain PO2 (phosphorescence quenching of oxyphor G4); cardiac output (CO) and renal and cerebral blood flow (ultrasound Doppler); hypoxia induced cellular responses (brain and kidney erythropoietin (EPO) mRNA and serum protein levels (ELISA)). Statistical analysis (SigmaPlot 14) was performed by ANOVA, Holm-Sidak and Mann-Whitney rank sum test when appropriate. Significance was assigned at p<0.05. Results In two models of anemia (hemodilution and RBC antibody mediated), acute reductions in blood CaO2 were associated with larger decreases in renal microvascular PO2, relative to brain microvascular PO2 (p<0.05). After acute hemodilution, there was a strong relationship between CaO2 and renal microvascular PO2 (r2=0.75). The magnitude of reduction in renal microvascular PO2 correlated with the degree of renal EPO mRNA expression and serum EPO protein levels. The magnitude of the increase in EPO mRNA was much larger in the kidney than in the brain (p<0.03). While no change in renal blood flow was observed in either model, a significant increase in common carotid and internal carotid blood flow was observed in both models (p<0.012). When DO2 was assessed, the kidney DO2 was reduced at all levels of anemia (p<0.01) whereas brain tissue DO2 was maintained in mild and moderate (Hb 90 and 70 g/L) (p=0.44) anemia but reduced in severe anemia (Hb 50 g/L) (p<0.02). The role of active cardiovascular increases in brain blood flow and maintained DO2 during anemia was impaired by systemic beta blockade, suggesting that active cardiovascular mechanisms are required to maintain optimal brain DO2 during anemia. Discussion Our analysis demonstrated: evidence of quantitative renal PO2 sensing of changes in CaO2; the clamping of renal blood flow (reduced DO2) during anemia may be a central mechanism allowing for sensing of changes in CaO2; reduced arterial CaO2 resulted in a local renal hypoxia response (increased serum EPO) and may have initiated the cardiovascular response to increase cerebral blood flow and maintain cerebral DO2. Inhibition of the adrenergic system impaired these responses and resulted in reduced brain DO2. Understanding the heterogeneous adaptive responses to acute anemia may inform clinical practice and optimize management of acutely anemia patients.

Antibacterial Activity of a New Ready-To-Use Calcium Silicate-Based Sealer.

The aim of this study was to evaluate the antibacterial potential of a calcium silicate-based sealer (Bio-C Sealer, Angelus) against common bacteria in primary and secondary endodontic infections. Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus mutans were exposed to fresh Bio-C Sealer for 24 h by the agar diffusion method (n=5). Additionally, the antibacterial activity was investigated against E. faecalis and S. mutans biofilms (48 h old) grown in discs with 4 mm in diameter and 2 mm in height. (n=3) of set discs of Bio-C Sealer (Angelus), EndoFill (Dentsply-Mallefer), Sealer 26 (Dentsply), AH Plus (Dentsply), Sealapex (Sybron-Endo) and EndoSequence BC Sealer (Brasseler). The antibacterial activity was evaluated by colony forming unity (CFU) counting using ImageJ software. Data were compared by one-way ANOVA followed by Holm-Sidak test (a=5%). Fresh Bio-C Sealer exhibited antimicrobial activity against all bacteria evaluated by agar diffusion method, except for S. mutans. Set discs of all endodontic sealers tested showed similar CFU values for E. faecalis (p>0.05). S. mutans in biofilms showed higher susceptibility to EndoFill compared with the other sealers (p<0.05). In conclusion, the results indicate that fresh Bio-C Sealer does not inhibit S. mutans growth, but exhibits antibacterial activity against E. faecalis, S. aureus, P. aeruginosa and E. coli. After setting, the Bio-C Sealer exhibits an antimicrobial potential comparable to that of the other sealers evaluated in E. faecalis biofilm, but lower than that of EndoFill for S. mutans biofilm.

Impact of ambient temperature on inflammation-induced encephalopathy in endotoxemic mice\u2014role of phosphoinositide 3-kinase gamma

BackgroundSepsis-associated encephalopathy (SAE) is an early and frequent event of infection-induced systemic inflammatory response syndrome. Phosphoinositide 3-kinase γ (PI3Kγ) is linked to neuroinflammation and inflammation-related microglial activity. In homeotherms, variations in ambient temperature (Ta) outside the thermoneutral zone lead to thermoregulatory responses, mainly driven by a gradually increasing sympathetic activity, and may affect disease severity. We hypothesized that thermoregulatory response to hypothermia (reduced Ta) aggravates SAE in PI3Kγ-dependent manner.MethodsExperiments were performed in wild-type, PI3Kγ knockout, and PI3Kγ kinase-dead mice, which were kept at neutral (30 ± 0.5 °C) or moderately lowered (26 ± 0.5 °C) Ta. Mice were exposed to lipopolysaccharide (LPS, 10 μg/g, from Escherichia coli serotype 055:B5, single intraperitoneal injection)—evoked systemic inflammatory response (SIR) and monitored 24 h for thermoregulatory response and blood–brain barrier integrity. Primary microglial cells and brain tissue derived from treated mice were analyzed for inflammatory responses and related cell functions. Comparisons between groups were made with one-way or two-way analysis of variance, as appropriate. Post hoc comparisons were made with the Holm–Sidak test or t tests with Bonferroni’s correction for adjustments of multiple comparisons. Data not following normal distribution was tested with Kruskal-Wallis test followed by Dunn’s multiple comparisons test.ResultsWe show that a moderate reduction of ambient temperature triggers enhanced hypothermia of mice undergoing LPS-induced systemic inflammation by aggravated SAE. PI3Kγ deficiency enhances blood–brain barrier injury and upregulation of matrix metalloproteinases (MMPs) as well as an impaired microglial phagocytic activity.ConclusionsThermoregulatory adaptation in response to ambient temperatures below the thermoneutral range exacerbates LPS-induced blood–brain barrier injury and neuroinflammation. PI3Kγ serves a protective role in suppressing release of MMPs, maintaining microglial motility and reinforcing phagocytosis leading to improved brain tissue integrity. Thus, preclinical research targeting severe brain inflammation responses is seriously biased when basic physiological prerequisites of mammal species such as preferred ambient temperature are ignored.

Intraarticular Adeno-Associated Virus Serotype AAV-PHP.S-Mediated Chemogenetic Targeting of Knee-Innervating Dorsal Root Ganglion Neurons Alleviates Inflammatory Pain in Mice.

Joint pain is the major clinical symptom of arthritis that affects millions of people. Controlling the excitability of knee-innervating dorsal root ganglion (DRG) neurons (knee neurons) could potentially provide pain relief. We undertook this study to evaluate whether the newly engineered adeno-associated virus (AAV) serotype, AAV-PHP.S, can deliver functional artificial receptors to control knee neuron excitability following intraarticular knee injection. The AAV-PHP.S virus, packaged with dTomato fluorescent protein and either excitatory (Gq ) or inhibitory (Gi ) designer receptors exclusively activated by designer drugs (DREADDs), was injected into the knee joints of adult mice. Labeling of DRG neurons with AAV-PHP.S from the knee was evaluated using immunohistochemistry. The functionality of Gq - and Gi -DREADDs was evaluated using whole-cell patch clamp electrophysiology on acutely cultured DRG neurons. Pain behavior in mice was assessed using a digging assay, dynamic weight bearing, and rotarod performance, before and after intraperitoneal administration of the DREADD activator, Compound 21. We showed that AAV-PHP.S can deliver functional genes into ~7% of lumbar DRG neurons when injected into the knee joint in a similar manner to the well-established retrograde tracer, fast blue. Short-term activation of AAV-PHP.S-delivered Gq -DREADD increased excitability of knee neurons in vitro (P = 0.02 by unpaired t-test), without inducing overt pain in mice when activated in vivo. By contrast, in vivo Gi -DREADD activation alleviated digging deficits induced by Freund's complete adjuvant-mediated knee inflammation (P = 0.0002 by repeated-measures analysis of variance [ANOVA] followed by Holm-Sidak multiple comparisons test). A concomitant decrease in knee neuron excitability was observed in vitro (P = 0.005 by ANOVA followed by Holm-Sidak multiple comparisons test). We describe an AAV-mediated chemogenetic approach to specifically control joint pain, which may be utilized in translational arthritic pain research.

Novel nanocarrier of miconazole based on chitosan-coated iron oxide nanoparticles as a nanotherapy to fight Candida biofilms

Overexposure of microorganisms to conventional drugs has led to resistant species that require new treatment strategies. This study prepared and characterized a nanocarrier of miconazole (MCZ) based on iron oxide nanoparticles (IONPs) functionalized with chitosan (CS), and tested its antifungal activity against biofilms of Candida albicans and Candida glabrata. IONPs-CS-MCZ nanocarrier was prepared by loading MCZ on CS-covered IONPs and characterized by physicochemical methods. Minimum inhibitory concentration (MIC) of the nanocarrier was determined by the microdilution method. Biofilms were developed (48 h) in microtiter plates and treated with MCZ-carrying nanocarrier at 31.2 and 78 μg/mL, in both the presence and absence of an external magnetic field (EMF). Biofilms were evaluated by total biomass, metabolic activity, cultivable cells (CFU), extracellular matrix components, scanning electron microscopy and confocal microscopy. Data were analyzed by two-way ANOVA and Holm-Sidak test (p < 0.05). A nanocarrier with diameter lower than 50 nm was obtained, presenting MIC values lower than those found for MCZ, and showing synergism for C. albicans and indifference for C. glabrata (fractional inhibitory concentration indexes of <0.12 and <0.53, respectively). IONPs-CS-MCZ did not affect total biomass and extracellular matrix. IONPs-CS-MCZ containing 78 μg/mL MCZ showed a superior antibiofilm effect to MCZ in reducing CFU and metabolism for single biofilms of C. albicans and dual-species biofilms. The EMF did not improve the nanocarrier effects. Microscopy confirmed the antibiofilm effect of the nanocarrier. In conclusion, IONPs-CS-MCZ was more effective than MCZ mainly against C. albicans planktonic cells and number of CFU and metabolism of the biofilms.

Cyclic contact fatigue resistance of ceramics for monolithic and multilayer dental restorations

ObjectiveTo evaluate the cyclic contact fatigue resistance and failure mode of ceramics for monolithic and multilayer restorations. MethodsCeramic structures (10 mm × 1.8 mm) were fabricated as follows (n = 28): (1) CAD-on- trilayer structure composed of Y-TZP (yttria stabilized tetragonal zirconia polycrystal- IPS e.max ZirCAD) infrastructure, fusion glass–ceramic (IPS e.max CAD Crystall/Connect) and lithium disilicate-based glass–ceramic (IPS e.max CAD); (2) ZFC- bilayer structure composed of Y-TZP infrastructure veneered by a fluorapatite glass–ceramic (IPS e.max Ceram); (3) LDC- monolithic lithium-disilicate glass–ceramic (IPS e.max CAD); and (4) YZW- monolithic Y-TZP (Zenostar Zr Translucent). All ceramics structures were bonded to a dentin analog substrate (G10). Specimens were submitted to cyclic contact fatigue test in a pneumatic cycling machine with 80 N load and 2 Hz frequency in distilled water at 37 °C. Test was interrupted after 104, 105, 5 × 105 and 106 cycles and the presence or absence of failure was recorded. Fatigue data were analyzed using Kaplan–Meier (log rank) and Holm–Sidak tests (α = 0.05). The relationship between the type of crack leading to failure and the experimental group was analyzed using chi-square test (α = 0.05). ResultsThere was no statistical difference between CAD-on and YZW groups (p = 0.516), which presented the highest survival rates after cyclic loading, followed by ZFC and LDC groups (p < 0.01). There was a significant relationship between type of crack and experimental group (p < 0.001). LDC specimens showed the greatest frequency of radial cracks, while cone cracks were more prevalent for ZFC and CAD-on specimens. SignificanceMonolithic Y-TZP (YZW) showed similar fatigue resistance to CAD-on multilayer specimens, but different failure mode. Monolithic lithium disilicate glass–ceramic (LDC) and Y-TZP conventionally veneered by glass–ceramic (ZFC) showed lower survival time under fatigue.

Porphyromonas Gingivalis Load is Balanced by 0.20% Chlorhexidine Gel. A Randomized, Double-Blind, Controlled, Microbiological and Immunohistochemical Human Study

Microbial contamination could compromise the stability of dental implants increasing the risk of inflammatory reactions in the surrounding soft tissues. In this human, randomized, double-blind, clinical study, the presence of Porphyromonas gingivalis on the healing abutment and the inflammatory infiltrate surrounding peri-implant soft tissues were investigated. Experiments were done in order to clarify the effect of 0.20% chlorhexidine (CHX) versus placebo, applied during each rehabilitation stage. Thirty patients (15 per group) were included. The load of adhering P. gingivalis on the healing screw were quantified by quantitative Polymerase Chain Reaction (qPCR) Taq-Man. Immunohistochemical analysis was carried out on the gingival biopsy. Moreover, clinical data were recorded. Analysis of variance and the Holm–Sidak test was used to evaluate differences between groups. The results showed a significant low presence of P. gingivalis load in healing abutments belonging to the 0.20% CHX group. Overall, the differences in terms of P. gingivalis DNA copy number between two groups were statistically significant (p < 0.01). All implants showed very low plaque and bleeding scores, but the placebo group appeared to have the highest expression of inflammation markers for T Lymphocytes, B Lymphocytes and macrophages Cluster definitions (CD3, CD20 and CD68). The use of 0.20% CHX could be recommended in all clinical procedures as it reduces significantly P. gingivalis load and host inflammatory response around implants.

Chondrotoxicity of Local Anesthetics: Liposomal Bupivacaine Is Less Chondrotoxic than Standard Bupivacaine.

Objective The purpose of this study is to determine whether (1) liposomal bupivacaine is chondrotoxic; (2) the chondrotoxicity of liposomal bupivacaine differs from standard bupivacaine; and (3) chondrotoxic effects are time dependent. Materials and Methods We obtained 72 10 mm articular cartilage plugs from 12 fresh bovine distal femoral knee joints and exposed them to either saline, 0.5% bupivacaine, or liposomal bupivacaine for either 30 or 90 minutes. Twenty-four hours after treatment, chondrocyte viability was measured with the use of a fluorescent live/dead assay. An ANOVA test of variance was performed followed by a Holm–Sidak test to make pairwise comparisons across conditions. Student's t-test was used to compare means. Results Percent viability of cells exposed to liposomal bupivacaine for 30 minutes was less versus saline control (53.9% ± 21.5% vs. 73.7 ± 18.4%, p=0.035), and this remained significant at 90 minutes (49.1% ± 20.3% vs. 67.2% ± 25.6%, p < 0.001). Liposomal bupivacaine had less chondrotoxic effects when compared with bupivacaine after 90 minutes, with greater viability (49.1% ± 20.3% vs. 21.4% ± 14.0%, p=0.003). Chondrotoxicity was found to be time dependent within the bupivacaine group (percent viability at 30 min: 45.5 ± 18.2%, 90 min: 21.4 ± 14.0%, p=0.001); however, liposomal bupivacaine did not demonstrate a significant time-dependent chondrotoxic relationship (p=0.583). Conclusions Bupivacaine and liposomal bupivacaine are both toxic to chondrocytes. Liposomal bupivacaine is less chondrotoxic than standard bupivacaine and does not demonstrate a time-dependent toxicity.

Anti-Predatory NIA for Unconstrained Mathematical Optimization Problems

Nature-Inspired Algorithms (NIAs) are one of the most efficient methods to solve the optimization problems. A recently proposed NIA is the anti-predatory NIA, which is based on the anti-predatory behavior of frogs. This algorithm uses five different types of self-defense mechanisms in order to improve its anti-predatory strength. This paper demonstrates the computation steps of anti-predatory for solving the Rastrigin function and attempts to solve 20 unconstrained minimization problems using anti-predatory NIA. The performance of anti-predatory NIA is compared with the six competing meta-heuristic algorithms. A comparative study reveals that the anti-predatory NIA is a more promising than the other algorithms. To quantify the performance comparison between the algorithms, Friedman rank test and Holm-Sidak test are used as statistical analysis methods. Anti-predatory NIA ranks first in both cases of “Mean Result” and “Standard Deviation.” Result measures the robustness and correctness of the anti-predatory NIA. This signifies the worth of anti-predatory NIA in the domain of mathematical optimization.

Bovine Heparin Demonstrates the Same Interaction with HIT Antibodies As Porcine Heparin

Heparin, an anticoagulant widely used in numerous medical applications, is considered an essential medicine by the WHO. Due to its high volume use and that it is the parent material for low molecular weight heparins, there is potential for the raw material to be in short supply. The African swine fever epidemic in China, ongoing since August 2018, has added further restraints on heparin source material supply. At present medical grade heparin in the US is only derived from porcine intestinal mucosa; however, there are explorations into using bovine, ovine, and other sources. Bovine heparin, once common place in the US pharmaceutical sector, is again under consideration by the US FDA. This study focused on the primary immunogenic activity associated with heparin, that is heparin-induced thrombocytopenia (HIT), and how the interaction of bovine heparin with functional HIT antibodies compares to that of porcine heparin. Materials and Methods Bovine unfractionated heparin from multiple manufacturers was compared to commercial medical grade porcine heparin obtained from US pharmacies. The US Pharmacopeia porcine heparin standard was used to determine potency equivalence. Antibodies to the complex of platelet factor 4/heparin (PF4/heparin) from banked clinically confirmed HIT patient apheresis fluids were combined with heparin and donor platelet rich plasma (PRP; blood collected in citrate from volunteers after signing a consent document). Heparins were tested at final concentrations of 0.1, 0.4, 0.8, 1, and 100 U/mL. The platelet activation response was determined on the BioData PAP-8 Platelet Aggregometer and quantitated in terms of primary slope (PS), area under the curve (AUC), maximum aggregation (MA), and final aggregation (FA). Characterization of the biophysical interaction between varying molar ratios of human PF4 and heparin was performed using photon correlation spectroscopy (PCS) and zeta potential (Zp) measurements of PF4/heparin complexes using Zetasizer Nano ZS instrumentation and software. Differences between bovine and porcine heparin were assessed by t-test or Mann-Whitney test. Concentration-response curves were analyzed by two-way ANOVA followed by the Holm-Sidak multiple comparison test using SigmaPlot software. Results Platelet activation to PF4/heparin antibodies at bovine and porcine heparin concentrations of 0.1 U/mL (56 ± 9 % vs. 54 ± 11 % MA) and 0.4 U/mL (59 ± 10 % vs. 65 ± 8 % MA) were the same with the expected inhibition (9 ± 4% MA) at the supra-therapeutic concentration of 100 U/mL. Consistent responses were obtained across 21 lots of bovine heparin, 30 lots of porcine heparin, and 38 platelet-HIT antibody combinations. The HIT potential of bovine heparin and porcine heparin was not statistically different (p&gt;0.05). At higher medical use doses, the platelet aggregation response in the presence of HIT antibodies was actually lower for bovine heparin than porcine heparin (0.8 U/mL, 49 ± 10 % vs. 64 ± 9 % MA, p&lt;0.05; and 1.0 U/mL, 45 ± 11 % vs. 62 ± 9 % MA, p&lt;0.05). By PCS, it was observed that the maximal aggregation between PF4 and either porcine or bovine heparin occurred at comparable molar ratios (7.3 ± 1.5 vs. 6.4 ± 0). Although the porcine and bovine heparins exhibited comparable molecular weights (16,333 ± 153 vs. 16,790 ± 230 Da) and polydispersities (1.19 ± 0.02 vs. 1.15 ± 0.01), porcine heparin formed somewhat larger complexes with PF4 (1113 ± 65 nm) than did bovine heparin (863 ± 68 nm). The molar ratios of PF4 to heparin at which the charge of the complex was fully neutralized (Zp = 0) was comparable for porcine and bovine heparin (9.04 ± 0.19 vs. 9.97 ± 0.65). Consistent responses were obtained across 4 lots of bovine heparin and 3 lots of porcine heparin. Conclusions Bovine heparin and porcine heparin had the same in vitro functional platelet activation response in the presence of HIT antibodies, the same potential to form complexes with human PF4, and the same associated features that make PF4 immunogenic. This investigation demonstrates that bovine heparins should have a similar immunogenic response as porcine heparin at equi-unit dosing. Current refinements in the manufacturing process for bovine and porcine heparins have led to well-characterized and purified products which is reflected in their comparable biological behavior. Disclosures No relevant conflicts of interest to declare.