Identification of APOB variants in samples from the brazilian population with hypercholesterolemic phenotype

BACKGROUND AND AIMS: Apolipoprotein B plays a crucial role in regulating plasma cholesterol by mediating the interaction of low-density lipoprotein (LDL) with LDL receptors in the liver. Inherited mutations in this gene may increase the risk of developing premature atherosclerotic cardiovascular disease, especially in individuals with familial hypercholesterolemia type 2 (FH2). The aim of this study is to identify APOB variants that may indicate pathogenicity in a sample of the Brazilian population using a data bank exome sequencing study by NGS in a Brazilian population phenotypically diagnosed by clinical and laboratory profile. This finding is going to improve genetic hypercholesteremia diagnosis. METHODS: High-quality DNA samples (n»300) were sequenced using an exon-targeted gene sequencing (ETGS) strategy to identify variants in FHrelated genes. Pathogenicity classification was based on criteria established by the American College of Medical Genetics and Genomics (ACMG), also using information from ClinVar and pathogenicity scores from previous association studies. RESULTS: A total of 121 variants were identified in APOB, of which four are novel variants missense (p.Thr626Asn, p.Ile2750Thr, p.Gln2078Lys and p.Met4184Arg). After curating pathogenicity scores, variants were classified according to the ACMG criteria. Among them four as pathogenic or likely pathogenic (p.Pro2739Leu, p.His1923Arg, p.Pro994Leu and p.Pro877Leu), and 21 variants had uncertain significance. Additionally, 92 previously known variants with uncertain significance were classified as benign or likely benign. The results were submitted to Clinvar for actualization of pathogenicity. CONCLUSIONS: These results improve the molecular diagnosis associating APOB variants with the clinical phenotype of hypercholesterolemia.

Effect of High-radiant Emittance and Short Curing Time on Polymerization Shrinkage Vectors of Bulk Fill Composites.

PURPOSE: To evaluate the effect of short curing time using a high-radiant emittance light on polymerization shrinkage vectors in different consistency bulk-fill composites (BFRCs) using micro-computed tomography. METHODS AND MATERIALS: Radiopaque zirconia fillers were homogeneously incorporated and functioned as radiopaque tracers into two regular-paste: TBFill (Tetric EvoCeram Bulk Fill) and TPFill (Tetric PowerFill), and two flowable (n=6): TBFlow (Tetric EvoFlow Bulk Fill) and TPFlow (Tetric PowerFlow) resin composites. Class I cavities (4 mm depth × 4 mm length × 4 mm width) were 3D-printed and filled in a single increment: TBFill and TBFlow were light-activated using a Bluephase Style 20i (10 seconds in high-mode); TPFill and TPFlow were light-activated using a Bluephase PowerCure (three seconds). The same adhesive system (Adhese Universal) was used for all groups. Microcomputed tomography scans were obtained before and after light-activation. Filler particle movement was identified by polymerization shrinkage vectors at five depths (from 0-4 mm): top, top-middle, middle, middle-bottom and bottom. RESULTS: TPFlow showed the lowest total vector displacement, followed by TBFlow, TBFill and TPFill, significantly different among each other (p<0.05). Generally, BFRCs showed decreased vector displacement with increased depth, and higher displacement at the top-surface (p<0.05). Qualitative analysis showed a similar pattern of vector magnitude and displacement for groups TBFill and TPFill, with displacement vectors on occlusal (top) surfaces toward the center of the restoration from the top to middle areas, and relatively limited displacement at the bottom. TBFlow and TPFlow showed more displacement on the occlusal (top). CONCLUSIONS: Short curing time with high-radiant emittance on fast-curing BFRCs was shown to be a feasible option in terms of vector displacement. Flowable BFRCs presented lower vector displacement than their regular-viscosity versions.

Effect of Argon Plasma Surface Treatment on Repair of Resin Composite Aged Two Years.

OBJECTIVES: To evaluate the effect of argon plasma treatment (PLA) when combined with sandblasting (SAN), silanization (SIL), and hydrophobic bonding resin (HBR) on the shear bond strength (SBS) of a two-year water-aged resin composite bonded to a newly placed composite after 24 hours and one year of water-storage. METHODS AND MATERIALS: Thirty-six light-cured composite plates (20mm x 20mm x 4mm thick) were obtained and stored at 37°C in distilled water for 2 years. These aged plates were distributed into 6 groups (n=6) according to the surface treatment: no treatment (Negative Control); SAN+SIL+HBR (Positive Control); SAN+PLA+SIL+HBR; PLA+ SIL+HBR; PLA+SIL; PLA+HBR. Fresh resin composite cylinders were built up using silicone molds (hole: 1.5 mm high x 1.5 mm diameter) positioned over the aged plates. Half of the SBS samples were stored in distilled water for 24 hours and loaded until failure, while the other half were stored for 1 year before being tested. Data were submitted to two-way analysis of variance and post-hoc Tukey Test (preset alpha of 0.05). RESULTS: Positive Control, SAN+PLA+SIL+HBR and PLA+SIL+HBR groups presented higher SBS means at the 24 hour evaluation. After 1 year of water storage, all groups demonstrated significant SBS reduction, with the SAN+PLA+SIL+HBR group presenting the highest SBS. CONCLUSIONS: Resin plasma treatment in combination with other surface treatments can improve the SBS of composite repairs after one year of water storage. The SBS of the composite repair was not stable over time regardless of the surface treatment.

Functional analysis of ldlr genetic variants identified in hypercholesterolemic patients in brazil, USING CRISPR/CAS9

BACKGROUND AND AIMS: Introduction: The familial hypercholesterolemia (FH) is one of the main causes of cardiovascular diseases, and it is mainly caused by genetic variants at the low-density lipoprotein receptor (LDLR). Although ultrasequencing technology has allowed the identification of several genetic variants, few of them was functional analyzed. The CRISPR/ Cas9 tool promotes precise genetic editing and allows the creation of experimental models, therefore contributing to the functional validation process. Aim: To use the CRISPR/Cas9 tool to perform in vitro functional analysis of LDLR variants identified in FH patients. METHODS: Two missense LDLR variants were selected within a group of variants identified in FH patients, based on in silico data, the affected protein domain and MAF. Three sgRNAs were designed for each of the variants c.551G>A and c.1118G>A, to analyze the accuracy of the sgRNAs. The sgRNAs were inserted on PX458 plasmid, cloned, purified in E. coli DH5a, and then co-transfected with the DNA template at HepG2 cells. The DNAs templates were designed to contain the selected variants. RESULTS: HepG2 cells co-transfected with PX458 constructs and DNA templates showed considerably transfection rate, being possible to visualize it at fluorescence microscopy. However, it was noted that single transfection of sgRNAs showed a higher transfection efficiency than cotransfection. CONCLUSIONS: We designed sgRNA for c.551G>A and c.1118G>A variants, being able to analyze the transfection efficiency. In further steps, we will select new sgRNAs for LDLR variants that have not been described yet, and functional analysis will be performed to determine the clinical relevance of these variants.

External Marginal Gap Evaluation of Different Resin-filling Techniques for Class II Restorations-A Micro-CT and SEM Analysis.

CLINICAL RELEVANCE: Secondary caries are the main reason for the failure of restorations, class II being the most affected. Techniques that promote less gap percentage are important. Flowable bulk fill composites used at such locations have been shown to decrease gap formation while being a faster procedure than an incremental technique.

Effect of Shortened Light-Curing Modes on Bulk-Fill Resin Composites.

CLINICAL RELEVANCE: Shortened light curing does not affect volumetric polymerization shrinkage or cohesive tensile strength but negatively affects the shear bond strength of some bulk-fill resin composites. When performing shortened light curing, clinicians should be aware of the light output of their light-curing units. SUMMARY: Purpose: To evaluate volumetric polymerization shrinkage (VPS), shear bond strength (SBS) to dentin, and cohesive tensile strength (CTS) of bulk-fill resin composites (BFRCs) light activated by different modes.Methods and Materials: Six groups were evaluated: Tetric EvoCeram bulk fill + high mode (10 seconds; TEC H10), Tetric EvoFlow bulk fill + high mode (TEF H10), experimental bulk fill + high mode (TEE H10), Tetric EvoCeram bulk fill + turbo mode (five seconds; TEC T5), Tetric EvoFlow bulk fill + turbo mode (TEF T5), and experimental bulk fill + turbo mode (TEE T5). Bluephase Style 20i and Adhese Universal Vivapen were used for all groups. All BFRC samples were built up on human molar bur-prepared occlusal cavities. VPS% and location were evaluated through micro-computed tomography. SBS and CTS tests were performed 24 hours after storage or after 5000 thermal cycles; fracture mode was analyzed for SBS.Results: Both TEC H10 and TEE H10 presented lower VPS% than TEF H10. However, no significant differences were observed with the turbo-curing mode. No differences were observed for the same BFRC within curing modes. Occlusal shrinkage was mostly observed. Regarding SBS, thermal cycling (TC) affected all groups. Without TC, all groups showed higher SBS values for high mode than turbo mode, while with TC, only TEC showed decreased SBS from high mode to turbo modes; modes of fracture were predominantly adhesive. For CTS, TC affected all groups except TEE H10. In general, no differences were observed between groups when comparing the curing modes.Conclusions: Increased light output with a shortened curing time did not jeopardize the VPS and SBS properties of the BFRCs, although a decreased SBS was observed in some groups. TEE generally showed similar or improved values for the tested properties in a shortened light-curing time. The VPS was mostly affected by the materials tested, whereas the SBS was affected by the materials, curing modes, and TC. The CTS was not affected by the curing modes.

Effect of Thermocycling on Biaxial Flexural Strength of CAD/CAM, Bulk Fill, and Conventional Resin Composite Materials.

New resin-based restorative materials have been developed, such as computer-aided design/computer-aided manufacturing (CAD/CAM) and bulk-fill composites, as an alternative to traditional layering techniques. This study evaluated the biaxial flexural strength (BFS) before and after thermocycling of five different resin composites: one hybrid resin/ceramic CAD/CAM indirect material, Lava Ultimate CAD-CAM Restorative (LU, 3M Oral Care); a conventional composite, Filtek Z350 XT (Z350, 3M Oral Care); two bulk-fill composites, Tetric N-Ceram Bulk Fill (TBF, Ivoclar Vivadent) and Filtek Bulk Fill (FBF, 3M Oral Care); and one bulk-fill flow resin composite, Filtek Bulk Fill Flow (FBFF, 3M Oral Care). Three hundred disc-shaped specimens (6.5 mm in diameter and 0.5 mm thick) were fabricated and divided into five groups (n=30 for each composite and condition). The BFS test was performed in a universal testing machine at a crosshead speed of 0.5 mm/min immediately (i, 24 hours) and after thermocycling (a, 500 thermal cycles of 5°C to 55°C with a 30-second dwell time). The Weibull modulus (m) and characteristic stress (η) were calculated, and a contour plot was used (m vs η) to detect differences between groups (95% two-sided confidence intervals). Significantly higher characteristic stress was observed for LUi (286.6 MPa) and Z350i (248.8 MPa) compared to the bulk-fill groups (FBFi=187.9 MPa, FBFFi=175.9 MPa, TBFi=149.9 MPa), with no differences between LUi and Z350i. Thermocycling significantly decreased the characteristic stress of all groups with the highest values observed for LUa (186.7 MPa) and Z350a (188.9 MPa) and the lowest for FBFFa (90.3 MPa). Intermediate values were observed for FBFa (151.6 MPa) and TBFa (122.8 MPa). The Weibull modulus decreased only for FBFa compared to FBFi. Composition and thermocycling significantly influenced the biaxial flexural strength of resin composite materials.

Análise funcional in silico das variantes no gene da APOB em pacientes com hipercolesterolemia familial / In silico functional analysis of variants in the APOB gene in patients with familial hypercholesterolemia

INTRODUÇÃO: O diagnóstico molecular da hipercolesterolemia familial (HF) é atribuído principalmente as variantes nos genes LDLR, LDLRAP1, APOB e PCSK9. O objetivo deste estudo foi realizar análise in silico para investigar o impacto de variantes sem descrição na literatura no gene APOB observado em pacientes com HF. MÉTODOS: Foram selecionados 141 indivíduos com diagnóstico clínico de HF. As variantes no gene da APOB foram selecionadas após sequenciamento dos éxons de 61 genes utilizando a plataforma MiSeq (Illumina). Os dados foram analisados nos programas Real Time Analysis, MiSeq Reporter, BaseSpace Sequence Hub e VariantStudio. Para a análise in silico, as sequências molde das moléculas da apoB-100 e o LDLr foram selecionadas por modelagem comparativa considerando o maior grau de identidade. As sequências proteicas foram alinhadas e os modelos 3D foram construídos utilizando os programas SEAVIEW e MODELLER v9.21. O gráfico de Ramachandran do modelo de menor energia apresenta 0,5% de outliers e análise de regiões de desordem, como principal validação. Os resultados das conformações de ancoragem foram analisados no software PyMol 2.1. Os estudos de docking molecular foram realizados para identificar o melhor complexo de conformação usando o servidor web clusPRO. RESULTADOS: Após a análise molecular dos 141 pacientes foram identificadas 7 variantes missenses sem descrição na literatura no gene APOB (c.433C>T, c.2630C>T, c.2950G>A, c.5743G>A, c.7367C>A, c.9880T>C e c.10780T>C). Os estudos de docking das variantes demonstraram uma maior afinidade entre o LDLr e a apoB-100 (c.2630C> T; Pro877Leu) em comparação com a proteína não mutada. A troca dos resíduos permaneceu como propriedade físico-química, e comparando as distâncias de ligação das proteínas não-mutadas (5Å) e mutadas (3,5Å), sugere-se uma maior afinidade do complexo (LDLr-apoB-100) para a leucina, tal fato é afirmado pela análise da região de desordens da apoB-100, onde a posição 877 está em uma região desorganizada e flexível. Esta maior afinidade poderia levar a uma menor dissociação intracelular deste complexo, resultando em uma alta taxa de degradação do LDLr pelas enzimas lisossômicas, levando ao aumento da concentração plasmática de LDLc. Para as outras variantes não houve alterações significativas. CONCLUSÃO: Os resultados sugerem que estudos in silico baseados na ferramenta de docking molecular podem melhorar o conhecimento da contribuição genética no desenvolvimento da doença HF. Além disso, a variante APOB c.2630C> T deve ser avaliada in vitropara validação do mecanismo proposto. (AU)

Effect of Argon Plasma Surface Treatment on Bond Strength of Resin Composite Repair.

OBJECTIVES:: This study evaluated the effect of argon plasma treatment (PLA) and its combination with sandblasting (SAN), silanization (SIL), and hydrophobic bonding resin (HBR) application on the micro-shear bond strength of water-aged restorative resin composite to a newly placed composite, simulating restoration repair. METHODS AND MATERIALS:: Forty-five light-cured composite plates (20-mm long × 20-mm wide × 4-mm thick) were fabricated using a hybrid composite and stored at 37°C in distilled water for six months. The aged composite surfaces were treated according to the following experimental groups, varying both treatment and order of application: 1) SAN + SIL + HBR (control), 2) SAN + PLA for 30 seconds + SIL + HBR, 3) SAN + SIL + PLA + HBR, 4) PLA + SIL + HBR, 5) PLA + SIL, 6) PLA + HBR, 7) SIL + PLA + HBR, 8) SIL + PLA, and 9) PLA. After the surface treatments, four fresh resin composite cylinders (1.5-mm high × 1.5-mm diameter) of the same composite were built on each aged composite surface using a silicone mold. After water storage for 24 hours or one year, the specimens were submitted to shear bond strength testing. Data were statistically analyzed by two-way analysis of variance and Tukey's test (5%). RESULTS:: Groups 1, 2, and 4 presented significantly higher bond strength means at 24 hours, although group 4 did not differ from group 7. Groups 5, 8, and 9 demonstrated significantly lower means than the other groups. Even though groups 1 and 2 had a significant bond strength reduction after 1 year, they still demonstrated higher bond strength at one year of storage. CONCLUSIONS:: While PLA application combined with surface treatment methods demonstrated high bond strength results, this treatment alone was not as beneficial as other methods that included SAN, SIL and HBR.

Assessment of range and quality of neck movement using a smartphone-based application.

This study had the objective of measuring the validity of using a smartphone-based application to measure range of motion (ROM) and quality of movement (QOM) of neck motion by comparing it with 3D-motion capture analysis. METHODS: Thirty healthy volunteers participated in this cross-sectional study. A helmet fitted with markers for motion capture analysis and a smartphone were fastened to the head of the participants. The smartphone recorded data using a beta version of Balancy (MEDEI, Denmark). Assessments of full active movement in transverse and sagittal planes were performed. Recordings were made simultaneously with the camera system and the smartphone. ROM and jerkiness were compared with a repeated measures ANOVA and a Pearson product moment was calculated to compare the outcomes from the different applications. Bland-Altman plots were generated to determine the levels of agreement. RESULTS: No difference was found between modalities when comparing measurements of jerkiness or ROM. An excellent Pearson product moment was found for the outcomes of the two modalities for ROM (Pearson's r: 0.83 - 0.96) and jerkiness (Pearson's r: 0.86 - 0.95). The Bland-Altman plot revealed a systemic offset where the phone consistently measured higher values for ROM and lower values for jerkiness. CONCLUSIONS: This study demonstrated that a smartphone-based application can be used to accurately measure ROM and jerkiness during neck movements. These results indicate the utility of using a smartphone-based application to assess neck movement in humans. The findings have implications for assessment of neck movement in research and clinical practice.

Increased postural stiffness during challenging postural tasks in patients with knee osteoarthritis with high pain sensitization.

BACKGROUND: Postural stability is affected in knee osteoarthritis patients who present with pain but the link to pain sensitization is unclear. METHODS: Patients with knee osteoarthritis completed the Knee Injury and Osteoarthritis Outcome Score and pressure pain thresholds were assessed bilaterally at the knee, lower leg and forearm prior to standing quietly (1 min) on a force platform in four conditions: Firm surface with open eyes, firm surface with closed eyes, soft surface with open eyes, and soft surface with closed eyes. Pain intensity during standing was assessed via numerical rating scale. Postural stability was assessed by the range, velocity, and standard deviation of the Center of Pressure (CoP) extracted from the force platform. The means of three repeated measures per standing condition were analysed. High-sensitization and low-sensitization groups were defined based on bilateral pressure pain thresholds from leg and arm. FINDINGS: Fifty-six patients were included. Compared with the low-sensitization group, the high-sensitization group demonstrated 1) smaller pressure pain thresholds at the knee (P < 0.05) although the Knee Injury and Osteoarthritis Outcome Score and pain intensity were not significantly different between groups, and 2) smaller range of the CoP in the anterior-posterior direction during the soft surface with closed eyes condition (P < 0.05). INTERPRETATION: Smaller CoP range suggest that patients with more widespread pain sensitivity have increased postural stiffness compared with the low-sensitization group. The greater stiffness found in high-sensitization patients under sensory restrictions (closed eyes and reduced proprioception) might relate to restricted integration of sensory information due to widespread pain sensitization.

Genomic analyses reveal two distinct lineages of Corynebacterium ulcerans strains.

Corynebacteriumulcerans is an important zoonotic pathogen which is causing diphtheria-like disease in humans globally. In this study, the genomes of three recently isolated C. ulcerans strains, 4940, 2590 and BR-AD 2649, respectively from an asymptomatic carrier, a patient with pharyngitis and a canine host, were sequenced to investigate their virulence potential. A comparative analysis was performed including the published genome sequences of 16 other C. ulcerans isolates. C. ulcerans strains belong to two lineages; 13 strains are grouped together in lineage 1, and six strains comprise lineage 2. Consistent with the zoonotic nature of C. ulcerans infections, isolates from both the human and canine hosts clustered in both the lineages. Most of the strains possessed spaDEF and spaBC gene clusters along with the virulence genes cpp, pld, cwlH, nanH, rpfI, tspA and vsp1. The gene encoding Shiga-like toxin was only present in one strain, and 11 strains carried the tox gene encoding the diphtheria-like toxin. However, none of strains 4940, 2590 and BR-AD 2649 carried any toxin genes. These strains varied in the number of prophages in their genomes, which suggests that they play an important role in introducing diversity in C. ulcerans. The pan-genomic analyses revealed a variation in the number of membrane-associated and secreted proteins that may contribute to the variation in pathogenicity among different strains.

Biofilm formation on different pH conditions by Streptococcus agalactiae isolated from bovine mastitic milk.

Streptococcus agalactiae is among the most relevant aetiologic agent of bovine clinical and subclinical mastitis, a major problem for the dairy industry. In Brazil, clonal diversity, capsular typing and multidrug resistance profiles of S. agalactiae related to human and bovine infections need further investigation. Presently, S. agalactiae isolates of bovine subclinical mastitis, from Brazilian Northeastern region, were submitted to capsular and pulsed-field gel electrophoresis (PFGE)-typing, antimicrobial susceptibility and assays of biofilm formation at different time incubation and pH levels. Sixteen bovine isolates were characterized by polymerase chain reaction assay as S. agalactiae capsular type II (CTII) and classified by PFGE in A1/A2 (n = 06), B1/B2 (n = 06), C (n = 03) and D (n = 01) patterns. Bovine S. agalactiae CTII strains were classified as 25% multidrug-resistant (MDR) with susceptibility to penicillin, linezolid and vancomycin. Biofilm formation on abiotic surface was strain- and time-dependent with significantly higher rates at pH 6·5. In conclusion, S. agalactiae capsular type II isolates recovered from bovine subclinical mastitis produced different pH-dependent biofilm levels. Our findings suggest that biofilm production is modulated by environmental factors and provides S. agalactiae advantageous in colonizing mammary gland during mastitis development, including MDR strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Streptococcus agalactiae is among the most relevant aetiologic agent of bovine clinical and subclinical mastitis, a major problem for the dairy industry. The disease may cause significant economic loss due to decreased production and milk quality and increased use of medicaments. Presently, data demonstrated that biofilm formation favours the establishment of infectious process in health mammary tissue by S. agalactiae and emphasizes that an acidic pH promotes adhesion by biofilm-forming bacterial strains. S. agalactiae strains (25%) showed resistance to tetracycline, azithromycin, erythromycin and clindamycin, and consequently were classified as multidrug-resistant strains.

Effect of Sonic Resin Composite Delivery on Void Formation Assessed by Micro-computed Tomography.

OBJECTIVES: The aim of this study was to quantify the internal void volume formation in commercially available, resin composites inserted using conventional or sonic insertion methods, and analyzed using three-dimensional (3D) micro-computed tomography (µCT). METHODS AND MATERIALS: Four resin composites were evaluated: one conventional (Herculite, Ultra, Kerr Corporation, Orange, CA, USA), one flowable bulk fill (SureFil SDR Flow, Dentsply International, York, PA, USA), and two packable bulk fill (SonicFill, Kerr Corporation, and Tetric EvoCeram Bulk Fill, Ivoclar Vivadent Inc, Schaan, Liechtenstein). Eight groups were evaluated according to each resin composite type and insertion method (conventional or sonic; n=5). Forty ABS 3D-printed cylindrical molds, 5.0 mm in diameter and 4.0 mm in depth, were fabricated. For the conventional resin composite, the mold was filled incrementally (two layers), while for bulk-fill resin composites, insertion was performed in a single increment. The sonic insertion method was performed using a specific handpiece (SonicFill Handpiece, Kerr Corporation). Resin composites were light cured using a multipeak light-emitting diode light-curing unit (VALO, Ultradent Products Inc, South Jordan, UT, USA) in its regular mode. Samples were evaluated by µCT, and data were imported into software (Amira, version 5.5.2, VSG, Burlington, MA, USA) for 3D reconstruction, from which the percentage of void volume was calculated. Data were analyzed using two-way analysis of variance and Tukey post hoc test at a preset alpha of 0.05. RESULTS: The conventional insertion method resulted in reduced porosity, compared with sonic insertion, for SureFil SDR Flow and Tetric EvoCeram bulk fill. The sonic insertion method did not demonstrate any influence on void formation for Herculite Ultra or SonicFill. CONCLUSION: Results suggest that the sonic insertion method might increase void formation during resin composite delivery, depending on restorative material brand.

Blood flow after contraction and cuff occlusion is reduced in subjects with muscle soreness after eccentric exercise.

Delayed onset muscle soreness (DOMS) occurs within 1-2 days after eccentric exercise, but the mechanism mediating hypersensitivity is unclear. This study hypothesized that eccentric exercise reduces the blood flow response following muscle contractions and cuff occlusion, which may result in accumulated algesic substances being a part of the sensitization in DOMS. Twelve healthy subjects (five women) performed dorsiflexion exercise (five sets of 10 repeated eccentric contractions) in one leg, while the contralateral leg was the control. The maximal voluntary contraction (MVC) of the tibialis anterior muscle was recorded. Blood flow was assessed by ultrasound Doppler on the anterior tibialis artery (ATA) and within the anterior tibialis muscle tissue before and immediately after 1-second MVC, 5-seconds MVC, and 5-minutes thigh cuff occlusion. Pressure pain thresholds (PPTs) were recorded on the tibialis anterior muscle. All measures were done bilaterally at day 0 (pre-exercise), day 2, and day 6 (post-exercise). Subjects scored the muscle soreness on a Likert scale for 6 days. Eccentric exercise increased Likert scores at day 1 and day 2 compared with day 0 (P<.001). Compared with pre-exercise (day 0), reduced PPT (~25%, P<.002), MVC (~22%, P<.002), ATA diameter (~8%, P<.002), ATA post-contraction/occlusion blood flow (~16%, P<.04), and intramuscular peak blood flow (~23%, P<.03) were found in the DOMS leg on day 2 but not in the control leg. These results showed that eccentric contractions decreased vessel diameter, impaired the blood flow response, and promoted hyperalgesia. Thus, the results suggest that the blood flow reduction may be involved in the increased pain response after eccentric exercise.

Altered pain sensitivity and axioscapular muscle activity in neck pain patients compared with healthy controls.

BACKGROUND: Previous studies have indicated that neck pain patients feel increased symptoms following upper limb activities, and altered axioscapular muscle function has been proposed as a contributing factor. METHODS: Pain sensitivity and muscle activity, during arm movements, were assessed in neck pain patients and controls. Patients with ongoing insidious-onset neck pain (IONP, N = 16) and whiplash-associated disorders (WAD, N = 9) were included along with sex- and age-matched controls (N = 25). Six series of repeated arm abductions were performed during electromyographic (EMG) recordings from eight bilateral muscles. The first and last three series were separated by 8 min and 42 s, respectively. Each series consisted of three slow and three fast movements. Pressure pain thresholds (PPTs) were recorded bilaterally from neck, head and arm at baseline, after the third and sixth movement series. Pain intensity was recorded on an electronic visual analogue scale (VAS). RESULTS: Larger pain areas and higher VAS scores were found in patients compared with controls (p < 0.001), and in patients, the VAS scores increased in the course of movements (p < 0.02). PPTs were lower in patients compared with controls at all sites (p < 0.03), and these decreased during arm movements in the IONP group (p < 0.03), while increasing at head and neck sites in controls (p < 0.04). During the slow movements, increasing serratus anterior EMG activity was found in the series with short breaks in-between for the WAD group compared with IONP and controls (p < 0.001). CONCLUSION: Axioscapular movement caused different responses in pain sensitivity and muscle activity between neck pain patient groups compared with controls. SIGNIFICANCE: Neck pain patients report increased symptoms following upper limb activities. This study shows that repeated arm movements caused differentiated responses in pain sensitivity and muscle activity between subgroups of neck pain patient and asymptomatic controls. Such findings may be of great clinical significance when planning rehabilitation for this patient population.

Experimental pain in the groin may refer into the lower abdomen: Implications to clinical assessments.

OBJECTIVES: To investigate the effects of experimental adductor pain on the pain referral pattern, mechanical sensitivity and muscle activity during common clinical tests. DESIGN: Repeated-measures design. METHODS: In two separate sessions, 15 healthy males received a hypertonic (painful) and isotonic (control) saline injection to either the adductor longus (AL) tendon to produce experimental groin pain or into the rectus femoris (RF) tendon as a painful control. Pain intensity was recorded on a visual analogue scale (VAS) with pain distribution indicated on body maps. Pressure pain thresholds (PPT) were assessed bilaterally in the groin area. Electromyography (EMG) of relevant muscles was recorded during six provocation tests. PPT and EMG assessment were measured before, during and after experimental pain. RESULTS: Hypertonic saline induced higher VAS scores than isotonic saline (p<0.001), and a local pain distribution in 80% of participants. A proximal pain referral to the lower abdominal region in 33% (AL) and 7% (RF) of participants. Experimental pain (AL and RF) did not significantly alter PPT values or the EMG amplitude in groin or trunk muscles during provocation tests when forces were matched with baseline. CONCLUSIONS: This study demonstrates that AL tendon pain was distributed locally in the majority of participants but may refer to the lower abdomen. Experimental adductor pain did not significantly alter the mechanical sensitivity or muscle activity patterns.

Reduced complexity of force and muscle activity during low level isometric contractions of the ankle in diabetic individuals.

BACKGROUND: This study evaluated the structure and amount of variability of surface electromyography (sEMG) patterns and ankle force data during low-level isometric contractions in diabetic subjects with different degrees of neuropathy. METHODS: We assessed 10 control subjects and 38 diabetic patients, classified as absent, mild, moderate, or severe neuropathy, by a fuzzy system based on clinical variables. Multichannel sEMG (64-electrode matrix) of tibialis anterior and gastrocnemius medialis muscles were acquired during isometric contractions at 10%, 20%, and 30% of the maximum voluntary contraction, and force levels during dorsi- and plantarflexion were recorded. Standard deviation and sample entropy of force signals were calculated and root mean square and sample entropy were calculated from sEMG signals. Differences among groups of force and sEMG variables were verified using a multivariate analysis of variance. FINDINGS: Overall, during dorsiflexion contractions, moderate and severe subjects had higher force standard deviation and moderate subjects had lower force sample entropy. During plantarflexion, moderate subjects had higher force standard deviation and all diabetic subjects had lower entropy. Tibialis anterior presented higher root mean square in absent group and lower entropy in mild subjects. For gastrocnemius medialis, entropy was higher in severe and lower in moderate subjects. INTERPRETATION: Diabetic neuropathy affects the complexity of the neuromuscular system during low-level isometric contractions, reducing the system's capacity to adapt to challenging mechanical demands. The observed patterns of neuromuscular complexity were not associated with disease severity, with the majority of alterations recorded in moderate subject.

Pain-evoked trunk muscle activity changes during fatigue and DOMS.

BACKGROUND: Muscle pain may reorganize trunk muscle activity but interactions with exercise-related muscle fatigue and delayed onset muscle soreness (DOMS) is to be clarified. METHODS: In 19 healthy participants, the trunk muscle activity during 20 multi-directional unpredictable surface perturbations were recorded after bilateral isotonic saline injections (control) and during unilateral and bilateral hypertonic saline-induced low back pain (LBP) in conditions of back muscle fatigue (Day-1) and DOMS (Day-2). Pain intensity and distribution were assessed by visual analogue scale (VAS) scores and pain drawings. The degree of fatigue and DOMS were assessed by Likert scale scores. Root-mean-square electromyographic (RMS-EMG) signals were recorded post-perturbation from six bilateral trunk muscles and the difference from baseline conditions (Delta-RMS-EMG) was extracted and averaged across abdominal and back muscles. RESULTS: In DOMS, peak VAS scores were higher during bilateral control and bilateral saline-induced pain than fatigue (p < 0.001) and during bilateral compared with unilateral pain (p < 0.001). The saline-induced pain areas were larger during DOMS than fatigue (p < 0.01). In response to surface perturbations during fatigue and DOMS, the back muscle Delta-RMS-EMG increased during bilateral compared with unilateral pain and control injections (p < 0.001) and decreased during unilateral pain compared with control injections (p < 0.04). In DOMS compared with fatigue, the post-perturbation Delta-RMS-EMG in back muscles was higher during bilateral pain and lower during unilateral pain (p < 0.001). The abdominal Delta-RMS-EMG was not significantly affected. CONCLUSION: Facilitated and attenuated back muscle responses to surface perturbations in bilateral and unilateral LBP, respectively, was more expressed during exercise-induced back muscle soreness compared with fatigue. SIGNIFICANCE: Back muscle activity decreased during unilateral and increased during bilateral pain after unpredictable surface perturbations during muscle fatigue and DOMS. Accumulation effects of DOMS on pain intensity and spreading and trunk muscle activity after pain-induction.