Genetic Polymorphisms in ACTN3 Contribute to the Etiology of Bruxism in Children.

Objective: Bruxism is a condition defined as a masticatory muscle activity with an unexplored genetic background. The aim of this study was to evaluate the association between genetic polymorphisms in ACTN3 and bruxism. Study design: A total of 151 biological-unrelated children, aged 7-12 years were included in a case control ratio of 1:1.5. The data collection was performed during interview and clinical examination. Saliva samples were collected from all children and 3 genetic polymorphisms in the ACTN3 (rs678397, rs1671064 and rs1815739) were selected for genotyping using real time PCR. Pearson chisquare calculation was used to assess Hardy-Weinberg equilibrium and to evaluate the association between genotypes and alleles frequencies for each genetic polymorphism in the co-dominant and recessive models. An alpha of 5% was used. Results: The genetic polymorphisms rs678397, rs1671064 and rs1815739 were associated with bruxism in the co-dominate model and in the recessive model (p<0.05). Allele distribution was also associated with bruxism for the polymorphisms rs678397 and rs1671064 (p<0.05). Conclusion: The genetic polymorphisms rs678397, rs1671064 and rs1815739 in ACTN3 are associated with bruxism and can contribute to the etiology of this condition in children.

Raman spectroscopy of stored red blood cell concentrate within sealed transfusion blood bags.

The standard practice in blood banks worldwide involves storage of red blood cells (RBCs) in plastic bags until they are needed for transfusion. During storage, the cells gradually degrade in functionality, a condition described as RBC storage lesion. Standard analytical techniques cannot assess the blood quality without breaching the sterility of the transfusion bag. In this study, we employed a commercially available spatially offset Raman spectroscopy (SORS) system using a custom designed protocol to non-invasively explore the biochemical changes in RBC concentrate of healthy donors over a storage period of approximately 42 days in standard transfusion bags, under standard storage conditions. The results reveal an increase in the oxygenation state of haemoglobin over the storage period for all donors, but different profiles for each donor. This study demonstrates the feasibility of acquiring consistent biochemical information relevant to the quality of stored blood, in situ through sealed blood transfusion bags using a commercially available instrument.

Impact of temporomandibular disorders on the stomatognathic system in children.

BACKGROUND: To evaluate the EMG activity and thickness of right masseter (RM), left masseter (LM), right temporal (RT) and left temporal (LT) muscles and bite force in children with temporomandibular disorders (TMD). MATERIAL AND METHODS: Forty five children (mean age 8.8 years; 22 boys and 23 girls) were examined on the basis of the RDC/TMD and the Faces Pain Scale-Revised (FPS-R) was used to determine the level of severity of the signs and symptoms of TMD, resulting in four groups: GI - without TMD (n=10); GII - with mild TMD (n=18), GIII: with moderate TMD (n=12) and GIV: with severe TMD (n=5). The data of electromyographic activity, maximum bite force and muscle thickness were tabulated and submitted to statistical analysis (ANOVA, P≤0.05). RESULTS: Children with TMD signs and symptoms had lower EMG activity than children of the control group. There was significant difference among the groups for the LT at rest (P=0.01), right (P=0.03) and left (P=0.05) laterality, and for the LM (P=0.01) and LT (P=0.03) muscles in maximum voluntary contraction. There were no statistically significant differences among the groups regarding muscle thickness. The bite force was lower in the TMD groups than children of the control group, with significant statistical difference for the right region (P=0.03). CONCLUSIONS: The severity of TMD signs and symptoms affected the EMG activity and the molar bite force in children. However, structural changes in the thickness of masticatory muscles are not perceptible in children with TMD signs and symptoms.

An investigation of red blood cell concentrate quality during storage in paediatric-sized polyvinylchloride bags plasticized with alternatives to di-2-ethylhexyl phthalate (DEHP).

BACKGROUND AND OBJECTIVES: Di-2-ethylhexyl phthalate (DEHP) is a blood bag plasticizer. It is also a toxin, raising concerns for vulnerable populations, for example, neonates and infants. Here, the in vitro quality of red cell concentrates (RCC) stored in paediatric bags formulated with alternative plasticizers to DEHP was compared. MATERIALS AND METHODS: RCC were pooled and split into polyvinylchloride (PVC)/DEHP, PVC/1,2-cyclohexanedicarboxylic acid diisononyl ester (DINCH) or PVC/butyryl trihexyl citrate (BTHC) bags. Quality was assessed on storage days 5, 21, 35 and 43. RESULTS: Metabolism differed among the bags: pCO2 levels were lowest and pO2 were highest in BTHC bags. Glucose consumption and lactate production suggested higher metabolic rates in BTHC bags. ATP levels were best maintained in DINCH bags (day 43 mean level: 2·86 ± 0·29 µmol/g Hb). RCC in BTHC bags had the greatest potassium release (54·6 ± 3·0 mm on day 43). From day 21, haemolysis was higher in BTHC bags (P < 0·01) and by day 43 had exceeded 0·8% (0·85 ± 0·10%). RCC in BTHC bags showed more microparticle formation than RCC in DEHP or DINCH bags. CONCLUSION: The results suggest that the BTHC formulation used was detrimental to RBC quality. DINCH bags could be a viable alternative to DEHP: they outperformed DEHP bags energetically, with better maintenance of ATP levels.

The effect of timing of gamma-irradiation on hemolysis and potassium release in leukoreduced red cell concentrates stored in SAGM.

While irradiation of red cell concentrates (RCC) prevents graft-versus-host disease in susceptible transfusion recipients, it also damages red blood cells (RBC). To understand the ability of irradiation regulations to prevent transfusion of inferior units, we irradiated 980 RCC in saline-adenine-glucose-mannitol (SAGM) using various combinations of pre-irradiation age and post-irradiation storage times, and measured hemolysis and extracellular potassium levels. We observed unacceptably high hemolysis (>0·8%) in some RCC and elevated extracellular potassium levels in all gamma-irradiated RCC. This suggests that more restrictive storage times should be considered for RCC in SAGM.

Standardization of CD62P measurement: results of an international comparative study.

BACKGROUND AND OBJECTIVES: Despite long being a mainstay in describing platelet activation via degranulation, interlaboratory variation remains an issue in measurement of membrane CD62P by flow cytometry. Our objective was to identify actions that may minimize this variation. MATERIALS AND METHODS: Sixteen laboratories participated in an international comparative study. Two sets of platelet samples were prepared in one laboratory. Set 1 was stained and fixed; set 2 was fixed and required staining at participating laboratories. A single-staining method was used, and platelet populations were selected based on forward scatter/side scatter characteristics. Calibration beads were used to standardize measurement across different instruments. RESULTS: There was a large discrepancy in reported CD62P values among study sites [interlaboratory coefficient of variance (CV): 36-78%]. When electronic data were re-analysed by a single analyst using a consistent gating strategy and a stable reference point, variation decreased markedly (CV < 12%), indicating a problem with isotype control samples, possibly related to sample fixation or shipment. CONCLUSION: Consensus regarding gating strategies and use of a reliable reference point would greatly improve agreement in interlaboratory CD62P measurement.

Prevalence of sleep bruxism in children: a systematic review of the literature.

The aim of the present investigation was to perform a systematic review of the literature dealing with the issue of sleep bruxism prevalence in children at the general population level. Quality assessment of the reviewed papers was performed to identify flaws in the external and internal validity. Cut-off criteria for an acceptable external validity were established to select studies for the discussion of prevalence data. A total of 22 publications were included in the review, most of which had methodological problems limiting their external validity. Prevalence data extraction was performed only on eight papers that were consistent as for the sampling strategy and showed only minor external validity problems, but they had some common internal validity flaws related with the definition of sleep bruxism measures. All the selected papers based sleep bruxism diagnosis on proxy reports by the parents, and no epidemiological data were available from studies adopting other diagnostic strategies (e.g. polysomnography or electromyography). The reported prevalence was highly variable between the studies (3·5-40·6%), with a commonly described decrease with age and no gender differences. A very high variability in sleep bruxism prevalence in children was found, due to the different age groups under investigation and the different frequencies of self-reported sleep bruxism. This prevented from supporting any reliable estimates of the prevalence of sleep bruxism in children.

Application of the ADVIA cerebrospinal fluid assay to count residual red blood cells in blood components.

BACKGROUND AND OBJECTIVES: There is no automated, accurate assay for the enumeration of residual red blood cells (rRBCs) in non-RBC components for transfusion, despite the potential risk of allo-immunization when mismatched components are transfused. MATERIALS AND METHODS: The automated ADVIA 120 cerebrospinal fluid (CSF) assay, which is approved to count RBCs and WBCs in CSF samples, was optimized and tested to measure rRBC in platelet concentrate (PC) and plasma components. RESULTS: Sample dilution, incubation time and reagent volume were optimized for use with non-RBC blood products. The assay was linear (R(2) = 0·99), even at low rRBCs counts. Intra- and inter-assay variation gave coefficients of variance (CV) between 2·2 and 9·4% and 2·6 and 14·9%, respectively, depending on rRBC levels. Good correlation (r = 0·995) was found between the automated assay and manual counting, which is considered the gold standard. Using the automated assay, the range of rRBCs (count/unit) in buffy-coat platelet concentrate (PCs) was 27-5505 × 10(6) and in apheresis PCs was 1-361 × 10(6). CONCLUSION: The ADVIA CSF assay is a sensitive, precise and accurate means to assess rRBC counts in non-RBC components.

A comparative study of common techniques used to measure haemolysis in stored red cell concentrates.

BACKGROUND AND OBJECTIVES: There is no standardized method of measuring the parameters for haemolysis determination of red cell concentrate (RCC). Three haemoglobin quantification methods (automated analyser, Harboe and Drabkin's) and two methods of haematocrit measurement (automated analyser and microcapillary centrifugation) were evaluated for use with RCC. MATERIALS AND METHODS: Twenty stored RCC were assayed for total haemoglobin, supernatant haemoglobin and haematocrit. RESULTS: Drabkin's and Harboe methods were linear (r(2) > or = 0.995) over 0.015-220 g/l haemoglobin. Overestimation by Drabkin's increased from 0% at 220 g/l to 137% at 0.015 g/l haemoglobin. Harboe values generally stayed within 6% of expected while haematology analyser values had a maximum 11% underestimation above 10 g/l. Analyser total haemoglobin was significantly lower (202 +/- 22 g/l) than Drabkin's (224 +/- 24 g/l) and Harboe (222 +/- 22 g/l) values. Haematocrit was greater via the analyser (65.7 +/- 5.7%) than with microcapillary centrifugation (59.3 +/- 5.7%). CONCLUSIONS: Harboe and Drabkin's methods are suitable for measuring total haemoglobin and supernatant haemoglobin in RCC. The analyser gave higher haematocrit values (11% on average) than did microcapillary centrifugation.

Liposome-complement interactions in rat serum: implications for liposome survival studies.

Serum complement opsonizes particles such as bacteria for clearance by the reticuloendothelial system. Complement has been reported to interact with liposomes and therefore may mediate the reticuloendothelial system clearance of liposomes. This study has used a rat serum model to define some of the characteristics of liposomes which modulate their ability to activate complement. Using functional hemolytic assays and C3/C3b crossed immunoelectrophoresis, we have demonstrated that liposomes activated rat complement in a dose-dependent manner with higher concentrations of liposomes activating higher levels of complement. The detection of complement activation required the inclusion of phospholipids bearing a net charge. Complement activation occurred via the classical pathway; no alternative pathway activation was detected. The presence of cholesterol contributed to complement activation in a dose-dependent manner. Phospholipid fatty acyl chain length did not influence complement activation while the introduction of unsaturated acyl chains markedly decreased levels of complement activation. Liposome size also influenced complement activation with 400 nm unilamellar vesicles more effectively activating complement than 50 nm vesicles for equivalent amounts of exposed lipid. These studies demonstrate that the composition of the liposome greatly affects the in vitro activation of rat serum complement and suggest that the biological half-life of liposomes in the circulation of rats may be altered by changing the liposome composition to reduce complement activation.

The effect of the Schwartz-Jampel syndrome on masticatory and facial musculatures--an electromyographic analysis.

This study had the goal to perform an electromyography evaluation of the orbicularis oris, orbicularis oculi, masseter, and temporal muscles of two siblings with Schwartz-Jampel syndrome (SJS), in different clinical activities, comparing them to healthy controls (C). The Schwartz-Jampel syndrome is a rare genetic disorder (71 cases reported in worldwide literature), in which myotonia may be observed in the facial muscles, determining a standard facie that shows an appearance of someone who is sad and weeping For the electromyography (EMG), a Myotronics--K61 electromyographer, was used, with superficial and disposable silver chloride electrodes. By means of the analysis of the obtained results, we observed significant statistical differences for the masseter muscle and for orbicularis oculi muscles (p < 0.01) among the studied groups, in which the individuals with SJS presented greater muscular activity than the normal ones, used as healthy controls. The statistical difference between the two groups was not significant for the temporal muscle, as well as for the orbicularis oris muscle, although the EMG averages were much greater in patients who were bringers of the syndrome. This high muscular activity may be related to the facial osseous alterations, evidenced in both patients with SJS, such as the mandibular migrognathia, with an atrophy of the mandibular cortex and a consequent approximation between the inferior dental root apices and the mandible base, as well as the presence of hypoplastic condyles in terms of size and height.

Rapid fibrin D-dimer tests for deep venous thrombosis: factors affecting diagnostic utility.

Measurement of fibrin D-dimer may be a useful diagnostic test to exclude a diagnosis of deep venous thrombosis (DVT) in the emergency department setting. However, the specific assay format may influence its sensitivity and ultimate clinical utility. We tested samples from 200 patients under evaluation for DVT using three fibrin D-dimer assays: the SimpliRED whole blood agglutination assay, a latex agglutination assay, and the Dimertest EIA. Latex agglutination assays were performed in both a specialized laboratory and a routine laboratory. The negative predictive value for all tests was > 90%. The sensitivity of the SimpliRED assay was similar to that of the latex assay. The sensitivity of the latex assay was significantly lower when performed by generalist laboratory technologists. Thus, while D-dimer may be a useful test for the exclusion of DVT, subjective endpoint latex agglutination assays should be performed only by appropriately trained personnel.

Recent updates on electrochemical degradation of bio-refractory organic pollutants using BDD anode: a mini review.

Boron-doped diamond (BDD) is playing an important role in environmental electrochemistry and has been successfully applied to the degradation of various bio-refractory organic pollutants. However, the review concerning recent progress in this research area is still very limited. This mini-review updated recent advances on the removal of three kinds of bio-refractory wastewaters including pharmaceuticals, pesticides, and dyes using BDD electrode. It summarized the important parameters in three electrochemical oxidation processes, i.e., anodic oxidation (AO), electro-Fenton (EF), and photoelectro-Fenton (PEF) and compared their different degradation mechanisms and behaviors. As an attractive improvement of PEF, solar photoelectro-Fenton using sunlight as UV/vis source presented cost-effectiveness, in which the energy consumption for enrofloxacin removal was 0.246 kWh/(g TOC), which was much lower than that of 0.743 and 0.467 kWh/(g TOC) by AO and EF under similar conditions. Finally the existing problems and future prospects in research were suggested.

Treatment of dilute methylene blue-containing wastewater by coupling sawdust adsorption and electrochemical regeneration.

In the present work, the coupling of adsorption and electrochemical oxidation on a boron-doped diamond (BDD) electrode to treat solutions containing dyes is studied. This coupling may be convenient for the treatment of diluted pollutant that is limited by the low rate of electrooxidation due to mass-transfer limitation. A pre-concentration step by adsorption could minimize the design of the electrochemical reactor. The adsorbent chosen was mixed with softwood sawdust, and methylene blue was chosen as the model dye molecule. Isotherms of adsorption and kinetics were investigated as well as the effects of current density and regeneration time. The BDD electrochemical oxidation of methylene blue adsorbed onto sawdust led simultaneously to its degradation and sawdust regeneration for the next adsorption. It was observed that multiple adsorption and electrochemical regeneration cycles led to an enhancement of adsorption capacity of the sawdust. This study demonstrated that adsorption­electrochemical degradation coupling offers a promising approach for the efficient elimination of organic dyes from wastewater.

Eleven new microsatellite loci for the tiger rattlesnake (Crotalus tigris).

Eleven microsatellite loci were isolated from an enriched genomic library from the tiger rattlesnake Crotalus tigris. Average observed heterozygosities in two populations were 0.456 and 0.427, respectively, and mean number of alleles were 7.54 (range 2-14) and 4.72 (range 2-13) respectively. No evidence of linkage disequilibrium was found across pairs of loci. The markers will be used in a long-term study examining the potential effects of urbanization on population dynamics and connectivity of this species in the mountain ranges surrounding Tucson, Arizona.

Effects of prestorage white cell reduction on platelet aggregate formation and the activation state of platelets and plasma enzyme systems.

BACKGROUND: The introduction of prestorage white cell (WBC) reduction in random-donor platelet concentrates in Canada has increased the occurrence of particulate material in PCs. The effects of filtration on platelet activation state and the activation of plasma enzyme systems were assessed. STUDY DESIGN AND METHODS: Particulate material was examined by light microscopy, electron microscopy, protein electrophoresis, and biochemical analysis. Thirty PCs (10 unfiltered, 20 filtered) were examined during processing and 5-day storage for pH, platelet count and mean volume, morphology, activation marker expression, and hypotonic shock response. Complement activation, thrombin generation, and fibrinolysis were assessed by using specific enzyme immunoassays or chromogenic assays. RESULTS: By all analyses, the particulate material appeared to be platelet aggregates. Platelets exposed to WBC-reduction filters expressed a significantly higher level of activation markers CD62 and CD63, altered morphology, and increased platelet microparticles throughout the storage period than did unfiltered platelets. Complement activation at the C3 level was significantly increased in filtered units with little evidence of coagulation or fibrinolytic system activation. CONCLUSION: Exposure of platelets to filters during prestorage WBC reduction increased platelet activation and mildly increased complement activation over the levels during the storage period. These alterations can contribute to the formation of irreversible platelet aggregates during processing.

Evaluación microbiológica y fisicoquímica de néctares pasteurizados elaborados con pulpa de tomate de árbol (Cyphomandra betaceae Sendth) / Microbiological and physicochemical evaluation of pasteurized nectar elaborated with tree tomato (Cyphomandra betaceae Sendth)pulp

Tree tomato (Cyphomandra betaceae Sendth) is a species from high tropical regions. In Venezuela, it is cultivate at the Andean and Aragua state but its consumption is restricted as fruit-fresh, though it a nutritious and industrial potential due its provitamin A content. In this research four nectars were elaborated in proportion I L pulp/4 L of water (1:4) and addition of ascorbic acid (I: 0 per cent; II: 0.5 per cent; III: 1.0 y IV: 1.5 per cent). The nectars were pasteurized (60 degrees C for 30 min), tuned into amber bottle, and stored under refrigeration conditions (7.0 +/- 1.0 degrees C). Weekly during 21 days the mesophilic bacteria, molds, yeasts, total coliforms (MPN/mL), pH, degree Brix, acidity, total carotenoids, vitamin C and total sugars were evaluated. The mesophilic bacteria content was < 200 UFC/mL for every formulation on the first day the evaluation, molds and yeasts were < 10 UFC/mL and the total coliforms (NPN/mL) < 3 until the third evaluation. No significant differences (P > 0.05) were founded on: pH. degree Brix and total sugars. Significant differences (P < 0.05) were founded in vitamin C and total carotenoids content. Sensorial analysis did not show significant differences between formulations for the smell and flavor attributes, when the color was discriminate during the evaluations. The formulation I (without acid ascorbic) had more preference due its color. In conclusion, the nectars showed useful life of 14-21 days under refrigeration condition storage due to the adequate physicochemical and microbiological quality of the product.