Local burn versus local cold induced acute effects on in vivo microcirculation and histomorphology of the human skin

Background: The impact of burns and colds on human skin microcirculation and histomorphology has not been compared as yet. Reflectance confocal microscopy (RCM) enables in vivo insight in human skin on cellular and subcellular levels. We evaluated analogies and differences of thermal injuries on microcirculation and histomorphology in vivo using RCM. Methods: Local superficial burn (6 female, 4 male; aged 28.4 ± 2.9 years, burn group) versus superficial cold (4 female, 6 male; aged 30.4 ± 5.2 years, cold group) was induced on the dorsum of the hand in an experimental immersion hand model. In vivo RCM was performed prior (control), immediately (t1) and 15 minutes (t2) following thermal injury to evaluate: Individual blood cell flow (IBCF), functional capillary density (FCD), epidermal thickness (ET), and granular cell size (GCS). Results: In the burn group, IBCF was increased at t1 (78.02 ± 2.60/min) and remained elevated at t2 (84.16 ± 3.04/min). In the cold group, IBCF decreased at t1 (12.62 ± 2.12 min) and increased at t2 (74.24 ± 3.14/min, P < 0.05) compared to the controls (58.23 ± 3.21/min). FCD was 6.74 ± 0.52/mm² in controls and increased at both t1 (7.82 ± 0.72/mm²) and t2 (8.02 ± 0.81/mm²) in the burn group. In the cold group, FCD decreased at t1 (2.60 ± 0.42/mm²) and increased at t2 (7.92 ± 0.44/mm², P < 0.05). ET increased at both t1 (43.12 ± 4.08 μm, P > 0.05) and t2 (47.26 ± 4.72 μm, P < 0.05) in the burn group. In the cold group, ET decreased at t1 (39.92 ± 3.14 μm, P > 0.05) and increased at t2 (44.72 ± 4.06 μm, P < 0.05) compared to the controls (41.26 ± 3.82 μm). Control GCS was 726.9 ± 59.4 μm² and increased at both t1 (739.8 ± 69.8 μm², P > 0.05) and t2 (762.6 ± 71.4 μm², P < 0.05) in the burn group. In the cold group, GCS decreased at t1 (712.4 ± 53.8 μm², P > 0.05) and increased at t2 (742.6 ± 64.8 μm², P < 0.05). Conclusions: Superficial burn induces more cellular destruction and cold leads to huge fluctuation in tissue perfusion, however, with moderate impact on histomorphology. The effect on dermal capillaries suggests a selective neural control and cold injuries might down‐regulate this system, much more than burns can activate it. Microsc. Res. Tech., 2011. © 2011 Wiley‐Liss, Inc.     

Biomechanical and microstructural benefits of physical exercise associated with risedronate in bones of ovariectomized rats

Several treatments have been developed aiming the prevention of bone loss. There are discussions about the best prophylactic and therapeutic procedures for osteoporosis. This study evaluated the effects of physical exercise associated with risedronate as a prophylactic and therapeutic procedure in osteopenic bones of rats submitted to ovariectomy. We used 48 Wistar rats divided into: ovariectomized or subjected to sham surgery. Ovariectomized rats were divided into the following sub‐groups: OVX, 12 weeks sedentary; OVX‐EX, treadmill training for 12 weeks; OVX‐RA, 12 weeks with risedronate administration; and OVX‐EX‐RA, 12 weeks with risedronate administration and treadmill training. Rats subjected to sham surgery were divided into the following sub‐groups: SH, 12 weeks sedentary; SH‐EX, treadmill training for 12 weeks; SH‐RA, 12 weeks with risedronate administration; and SH‐EX‐RA, 12 weeks with risedronate administration and training on the treadmill. The effectiveness of the treatment was evaluated in tibias using biomechanical, radiological, histomorphometric, and immunohistochemical analyses. Data were analyzed by statistical tests, with significance level of P < 0.05. Results of mechanical tests showed that the SH‐RA group had lower values compared with OVX‐RA group; densitometry showed no significant differences; according to histomorphometric methods, OVX group presented lower results than the SH‐EX, OVX‐RA, SH‐EX‐RA, and OVX‐EX‐RA groups, and SH‐EX‐RA and OVX‐EX‐RA groups showed values higher than SH‐RA, SH, and OVX‐EX groups. The SH‐EX‐RA and OVX‐EX‐RA groups had decreased immunostaining for tartrate‐resistant acid phosphatase and receptor activator of nuclear factor kappa‐B ligand and increased osteoprotegerin immunostaining. In this experimental model, it was concluded that the physical training associated with use of risedronate exerted positive effects on biomechanical and microstructural properties in bones of ovariectomized rats. Microsc. Res. Tech. 77:431–438, 2014. © 2014 Wiley Periodicals, Inc.     

Automated identification of neurons and their locations

Individual locations of many neuronal cell bodies (>10⁴) are needed to enable statistically significant measurements of spatial organization within the brain such as nearest‐neighbour and microcolumnarity measurements. In this paper, we introduce an Automated Neuron Recognition Algorithm (ANRA) which obtains the (x, y) location of individual neurons within digitized images of Nissl‐stained, 30 μm thick, frozen sections of the cerebral cortex of the Rhesus monkey. Identification of neurons within such Nissl‐stained sections is inherently difficult due to the variability in neuron staining, the overlap of neurons, the presence of partial or damaged neurons at tissue surfaces, and the presence of non‐neuron objects, such as glial cells, blood vessels, and random artefacts. To overcome these challenges and identify neurons, ANRA applies a combination of image segmentation and machine learning. The steps involve active contour segmentation to find outlines of potential neuron cell bodies followed by artificial neural network training using the segmentation properties (size, optical density, gyration, etc.) to distinguish between neuron and non‐neuron segmentations. ANRA positively identifies 86 ± 5% neurons with 15 ± 8% error (mean ± SD) on a wide range of Nissl‐stained images, whereas semi‐automatic methods obtain 80 ± 7%/17 ± 12%. A further advantage of ANRA is that it affords an unlimited increase in speed from semi‐automatic methods, and is computationally efficient, with the ability to recognize ～100 neurons per minute using a standard personal computer. ANRA is amenable to analysis of huge photo‐montages of Nissl‐stained tissue, thereby opening the door to fast, efficient and quantitative analysis of vast stores of archival material that exist in laboratories and research collections around the world.     

Influence of different calcium hydroxide removal protocols on the bond strength of epoxy resin-based sealer in long oval root canals

This study evaluated the effect of different protocols for Ca(OH)₂ removal on the bond strength (BS) of epoxy resin-based sealer in long oval root canals. Forty-eight mandibular incisors with long oval root canals were selected by CBCT. Biomechanical preparation was performed, the samples were filled with Ca(OH)₂ paste and distributed in four groups (n = 12): G-without Ca(OH)₂ paste (C); G2-syringe/needle (SN); G3-SN + XP-Endo® Finisher (XPF); G4-SN + passive ultrasonic irrigation (PUI). After 7 days, Ca(OH)₂ was removed and teeth were filled using AHPlus and guttapercha. In four samples from each group, the sealer was manipulated with 0.1% rhodamine to assess sealer penetration by fluorescent laser confocal microscopy (FLCM). Samples were sectioned in slices for BS, scanning electron microscopy, and FLCM analysis. The BS data were analyzed by ANOVA and Tukey tests (α = 0.05). The lowest BS values were found for the SN group (12.89 ± 4.36) compared to C (35.55 ± 10.05), while PUI group presented intermediate values (28.57 ± 9.35) and the XPF group (31.34 ± 9.8) showed values that were sometimes similar to C group and sometimes similar to PUI group (p > .05). The analysis of the adhesive interface showed evident gaps, with the presence of residues between the dentin and the filling material for the SN group, and for C, XPF and PUI groups juxtaposed adhesive interface. FLCM images showed sealer penetration in the dentinal tubules along the entire perimeter for C, XPF, and PUI groups. Ca(OH)₂ removal with XPF and PUI from long oval root canals resulted in higher bond strength values compared to SN, besides better sealer penetration on dentinal tubules and juxtaposed adhesive interface.     

A rabbit maxillary sinus model with simultaneous customized‐implant placement: Comparative microscopic analysis for the evaluation of surface‐treated implants

We describe the use of a rabbit maxillary sinus model, characterized by thin osseous tissue and low bone density, for the evaluation of surface‐treated implants by histologically and histomorphometrically comparing the osseointegration patterns depending on the surface treatment methods. Twenty rabbits were randomly assigned to two groups of 10 animals, one receiving 5 × 3 mm customized implants (machined, MA or sandblasted and acid etched, SLA) placed in sinus and the other receiving implants placed in a tibia. Histological observation of the implant placed in sinus shows relatively more active new bone formation, characterized by trabecular bone pattern underneath the cortical bone in sinus as compared with that in tibia. Histomorphometric analysis in the rabbits receiving implants in a tibia, the NBIC (%) associated with the SLA surface implant was greater than that associated with the MA implant at 2 weeks (55.63 ± 8.65% vs. 47.87 ± 10.01%; P > 0.05) and at 4 weeks (61.76 ± 9.49% vs. 42.69 ± 10.97%; P < 0.05). Among rabbits receiving implants in a sinus, the NBIC (%) associated with the SLA surface implant was significantly greater than that associated with the MA surface implant both at 2 weeks (37.25 ± 7.27% vs. 20.98 ± 6.42%; P < 0.05) and at 4 weeks (48.82 ± 6.77% vs. 31.51 ± 9.14%; P < 0.05). As a result, we suggest that the maxillary sinus model is an appropriate animal model for assessing surface‐treated implants and may be utilized for the evaluation of surface‐treated implants in poor bone quality environment. Microsc. Res. Tech. 78:697–706, 2015. © 2015 Wiley Periodicals, Inc.     

Micro‐Computed Tomography and Scanning Electron Microscopy Comparisons of Two Nickel–Titanium Rotary Root Canal Instruments Used With Reciprocating Motion

The single‐file root canal instrumentation technique using reciprocating motion has been gaining concern. Therefore, the purpose of this study was to compare the shaping ability of single ProTaper F2 file and WaveOne Primary file when they were used in the curved root canal with reciprocation motion and to investigate the durability of the file after use with a scanning electron microscopy (SEM). Changes in structure model index (SMI), root canal volume, curvature, surface area, and degree of transportation were measured from the cross‐sectional images of the prepared canals using the micro‐CT system with an isotropic resolution of 16 μm. Results showed that there were no differences in the changes of root canal volume, surface area, and SMI between the two file groups after the preparation (p > 0.05). The ProTaper group showed a curvature straightening value of 25.45 ± 12.51%, while the WaveOne group showed 27.30 ± 10.91%, and there was no statistically significant difference (p > 0.05). The transportation values between the two groups showed no significant differences (p > 0.05). SEM revealed that 60% of ProTaper files showed initiation of microcracks on the surface while those were detected on the only one WaveOne file. The single‐file technique using either WaveOne Primary or ProTaper F2 can be safely used under each reciprocating motion without creating an increased apical transportation in curved canals. However, the metallurgic property resists cyclic fatigue was more favorable with WaveOne under the scanning evaluation. SCANNING 35: 112‐118, 2013. © 2012 Wiley Periodicals, Inc.     

Morphological,microscopic, and physicochemical studies of Diospyros montana

Adulteration is the root cause of producing not only a chemically and pharmacologically inferior but also in some instances hazardous or poisonous drug. Despite availability of several techniques, microscopy and physicochemical analyses are the most practical approaches for crude drug authentication. Hence, the present study aimed to evaluate morphological, microscopic, and physicochemical properties of root, bark, leaf, and fruit of Diospyros montana Morphological properties were determined by sensory organs, whereas microscopic features of cross‐sections and powders were determined by light and scanning electron microscopy. The proximate and fluorescence analyses were performed using the standard guidelines. The physical examination of fresh, shade‐dried, and powdered material showed no significant change in color. The identifying cellular structures included cuboidal cork, pitted tracheids, scalariform, reticulate and spiral xylary vessels, and rosettes, raphide, and cuboidal calcium oxalate crystals. The stomatal number, stomatal index, vein‐islet and vein‐termination number, and palisade ratio in the leaf were found to be 293.91 ± 32.68 mm^?2, 64.18 ± 3.42%, 22.00 ± 3.81 mm^?2 and 38.40 ± 5.81 mm^?2, and 3.85 ± 0.60, respectively. Total ash, acid insoluble ash, water soluble ash and sulfated ash of leaf (9.00 ± 0.50%, 1.67 ± 0.23%, 2.00 ± 0.22% and 14.50 ± 0.99%, respectively), foaming index of bark and root (111.11 ± 2.11), and swelling index of fruit (19.00 ± 3.45) were higher than the other parts. The powder of different parts showed characteristic colors in the daylight and UV light upon treatment with various regents. The plant was found to be rich in saponins, fibers, and flavonoids. The results of the present study may serve as identifiers of different parts of Diospyros montana.     

Reflectance confocal‐laser‐scanning microscopy in vivo assessments of cigarette‐induced dynamic alterations of cutaneous microcirculation on histomorphological level

Objective: Until now, high resolution reflectance confocal‐laser‐scanning microscopy (CLSM) was used for observation of cutaneous morphology in vivo and in real time. We hypothesized that CLSM also allows observation of dynamic processes of cutaneous microcirculation. Methods: Reflectance CLSM (Vivascope1500; Lucid, Rochester, NY) was performed in 24 young male habitual smokers (23 years, range: 19–26, body mass index 23.9 ± 4.04) with relatively limited cigarette exposure (mean: 3.1 ± 2.4 pack‐years). Eight matched nonsmokers served as controls. The quantitative blood cell flow and the diameter of capillary loops were determined prior (baseline), during, as well as 5 and 10 min after smoking. Results: Baseline value for blood cell flow was 55.50 ± 2.33 cells/min, and decreased over 45% during smoking (30.43 ± 3.76/min; P = 0.02). They were still 22% lower (43.33 ± 2.45/min; P = 0.01) 5 min after smoking and exceeded baseline values 10 min after smoking by 13% (63.00 ± 3.10/min; P > 0.05). The baseline values for capillary loop diameter (9.03 ± 0.22 μm) decreased by 21% (7.18 ± 0.28 μm; P = 0.03) during smoking, remained about 9% (8.23 ± 0.18 μm; P = 0.01) lower 5 min after smoking and exceeded baseline values insignificantly by 4% (9.38 ± 0.28 μm; P > 0.05) 10 min after smoking. There were no significant differences to the controls. Conclusion: Reflectance CLSM enables qualitative and quantitative observation of dynamic processes of cutaneous microcirculation on histomorphological level. Microsc. Res. Tech., 2009. © 2008 Wiley‐Liss, Inc.     

腔镜下食管癌根治术在食管癌治疗中的应用

目的探讨应用胸腔镜联合腹腔镜进行食管癌根除术的可行性和疗效。方法选取2009年9月至2011年6月入住我院的食管癌患者42例,完全随机分为微创组(胸腔镜联合腹腔镜食管癌根治组)21例、开放组(传统开放食管癌根治组)21例。比较两组围手术期临床资料、淋巴结清扫数及阳性转移率及主要并发症情况。结果微创组平均手术时间265±57min与开放组362±83min间相比较差异不具有统计学意义(P=0.11>0.05);微创组平均手术出血量187±21ml与开放组279±38ml间相比较差异具有统计学意义(P=0.04<0.05);微创组平均住院天数10±3dl与开放组16±2d之间相比较差异具有统计学意义(P=0.04<0.05)。微创组平均清扫淋巴结14±4个与开放组13±5个间相比较差异不具有统计学意义(P=0.80>0.05);微创组术后转移率为38.1%(8/21)与开放组42.9%(9/21)相比较差异不具有统计学意义(P=0.75>0.05)。开放组术后出现肺部感染1例(4.8%),肺不张1例(4.8%),而微创组术后未出现肺部感染及肺不张病例,与开放组比较差异具有统计学意义(P=0.049<0.05)。结论胸腔镜联合腹腔镜食管癌根治术及胸、腹腔淋巴结清扫治疗食管癌安全可行且具有出血量小、住院天数少、术后并发症少等优势。     

Histological and histochemical evaluations on the effects of high incubation temperature on the embryonic development of tibial growth plate in broiler chickens

The effects of experimentally induced high incubation temperature on the embryonic development of growth plate of the chicken were investigated by means of histological and enzyme histochemical methods. In the experiments, 250 fertile eggs of Ross‐308 broiler strain were divided into two groups, the control eggs were maintained under optimal conditions (37.8°C and 65% ± 2% relative humidity, rh) during the whole incubation period. Heat‐stress imposed eggs were maintained under normal conditions (37.8°C and 65% ± 2% rh) until the 10th day of incubation, and then, continuously (24 h per day) exposed to high temperature (38.8°C and 65% ± 2% rh). Tissue samples were taken from 10 animals of each group at the 11th, 13th, 15th, 18th, 21st days of incubation. Tissue samples were processed by enzyme histochemical methods in addition to routine histological techniques. The relative tibia weights and tibia length were lower in the heat‐stress group compared to the control group. The results of the measurements of the growth plate showed that the proliferative zone was narrowed whereas, the transitional and hypertrophic zone were thickened in the heat stress group. Alkaline phosphatase (ALP) density was significantly decreased in the heat‐stress group compared to the control group. In conclusion, bone formation and growth plate formation are crucial for embryo development and 1°C higher from optimum may increase the incidence of skeletal disorders and leg problems in broiler chickens which is one of the major animal welfare concerns for the poultry industry. Microsc. Res. Tech. 79:106–110, 2016. © 2016 Wiley Periodicals, Inc.     

Quantitative evaluation of root canal surface roughness after filing with adaptive reciprocating and continuous rotary instruments

Obtaining clean and smooth root canal walls is the ideal clinical outcome of the cleaning and shaping stage in root canal treatment. This study compares the surface roughness of root canal surfaces instrumented with a NiTi filing system with either adaptive reciprocating (AR) or continuous rotation (CR). Root canal cleaning and shaping was carried out on the mesial canals of 24 extracted first molars roots with either AR or CR. Roots were split in half and the surface roughness of their canals was evaluated in 12 three dimensional roughness reconstructions using a scanning electron microscope. Rz (nm) values were calculated in three areas of each reconstruction and analyzed (α = 0.05). Mann‐Whitney tests showed that surface roughness was significantly higher overall in the AR group (Rz = 967 ± 250 nm) compared with the CR group (Rz = 739 ± 239 nm; p = 0.044). The roughness values generally increased from apical towards the coronal third in both groups. A less aggressive finishing file or a continuous rotary system to end the cleaning and shaping stage may be beneficial to reduce roughness of the root canal surface.     

MicroRNA expression profile and lipid metabolism characteristics in liver of rat undergoing high-fat diet

This study aimed to investigate the microRNA expression profile and the characteristics of lipid metabolism in the livers of rats undergoing a high-fat diet. Fifty male Sprague-Dawley (SD) rats were divided into a standard chow group (C group, N = 10) and a high-fat diet group (H group, N = 40). After 12 weeks, the rat body weight, body length, fat mass, and serum lipid concentration were measured. The expression profile of microRNAs and the gene and protein expression levels involved in lipid metabolism in rat liver were detected. Body fat and serum lipid concentrations were all significantly higher in the H group than those in the C group (p < 0.05 or p < 0.01). The expression of 10 microRNAs showed significant differences in the liver (p < 0.05). In particular, the let-7 family expression levels significantly increased (p < 0.05) in the H group compared with those in the C group. Compared with the C group, the high-fat diet resulted in low FAS, CPT1A, and ApoAI mRNA expression levels (p < 0.05 or p < 0.01) and high PPARα and FAT/CD36 mRNA expression levels in the H group rat liver (p < 0.01). Meanwhile, the protein PPARα, FAS, CPT1A, FAT/CD36, and ApoAI expression levels were all significantly lower in the H group than those in the C group (p < 0.05 or p < 0.01). In conclusion, the high-fat diet increased the body fat and serum lipid levels and altered the 10 microRNA expression levels in the liver. The high-fat diet may affect hepatic carbohydrate metabolism and increase ectopic fat accumulation through let-7 family overexpression. The high-fat diet for 12 weeks decreased lipid metabolism level in the liver, thereby decreasing fatty acid synthesis, oxidation, and transport by down-regulating the PPARα, FAS, CPT1A, FAT/CD36, and ApoAI protein levels.     

Effect of processing methods for transmission electron microscopy on corneal collagen fibrils diameter and spacing

Introduction: The corneal tissue was processed in fixatives and embedded in resin for transmission electron microscopy to observe the ultrastructure of the collagen fibrils (CFs). The effect of these processing methods on the CF diameter and the interfibrillar spacing was studied. Methods: Four normal human corneal buttons were used for this study. A part of each cornea was fixed in 2.5% glutaraldehyde containing cuprolinic blue in sodium acetate buffer and embedded in spurr's resin (SpurrCB). A second part of each cornea was fixed in 2.5% glutaraldehyde + osmium tetroxide and embedded spurr's resin (SpurrOsm). The third part of each cornea was fixed in paraformaldehyde (4%) and embedded in LR White at 4°C (LRWhite). Ultrathin sections were stained with uranyl acetate and lead citrate. Results: In the tissue, fixed in SpurrCB, the diameter was 38.4 ± 5.9 nm and spacing between CF was 52.5 ± 5.3 nm. In the tissue fixed in SpurrOsm, the diameter was 28.37 ± 5.84 nm and spacing between CF was 45 ± 4.57 nm. In the tissue fixed in LR White, the CF diameter was 24 ± 2.3 nm and spacing between CF was 39.0 ± 4.2 nm. The diameters and interfibrillar spacing of the tissue processed by SpurrCB, SpurrOsm, and LRWhite were significantly different (P < 0.001) from one another. Conclusion: Our study shows that there is a variation in the CF diameter and spacing depending on the method of fixation and embedding resins used. This needs to be considered when comparative studies using different methods are done. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Antimicrobial effect,frictional resistance,and surface roughness of stainless steel orthodontic brackets coated with nanofilms of silver and titanium oxide: a preliminary study

Nano‐silver and nano‐titanium oxide films can be coated over brackets in order to reduce bacterial aggregation and friction. However, their antimicrobial efficacy, surface roughness, and frictional resistance are not assessed before. Fifty‐five stainless‐steel brackets were divided into 5 groups of 11 brackets each: uncoated brackets, brackets coated with 60 µm silver, 100 µm silver, 60 µm titanium, and 100 µm titanium. Coating was performed using physical vapor deposition method. For friction test, three brackets from each group were randomly selected and tested. For scanning electron microscopy and atomic‐force microscopy assessments, one and one brackets were selected from each group. For antibacterial assessment, six brackets were selected from each group. Of them, three were immediately subjected to direct contact with S. mutans. Colonies were counted 3, 6, 24, and 48 h of contact. The other three were stored in water for 3 months. Then were subjected to a similar direct contact test. Results pertaining to both subgroups were combined. Groups were compared statistically. Mean (SD) friction values of the groups 'control, silver‐60, silver‐100, titanium‐60, and titanium‐100' were 0.55 ± 0.14, 0.77 ± 0.08, 0.82 ± 0.11, 1.52 ± 0.24, and 1.57 ± 0.41 N, respectively (p = .0004, Kruskal–Wallis). Titanium frictions were significantly greater than control (p < .05), but silver groups were not (p > .05, Dunn). In the uncoated group, colony count increased exponentially within 48 h. The coated groups showed significant reductions in colony count (p < .05, two‐way‐repeated‐measures ANOVA). In conclusions, all four explained coatings reduce surface roughness and bacterial growth. Nano‐titanium films are not suitable for friction reduction. Nano‐silver results were not conclusive and need future larger studies.     

Determination of protoberberine alkaloids in Coptidis Rhizoma by novel extraction and high-performance liquid chromatography

An alternative, effective, and solvent-free microextraction method has been developed using weighing paper as the adsorbent for five protoberberine alkaloids from the methanolic extract of Coptidis Rhizoma (C. Rhizoma). Several variables influencing extraction efficiency were optimized. 1.0?×?1.0?cm glassine weighing paper was employed for the isolation of the analytes followed by high-performance liquid chromatography. Under the optimum conditions, the limits of detection were between 0.2 and 0.4?ng/mL with enrichment factors from 41–47. The calibration curves were linear across the concentration ranges examined with correlation coefficients exceeding 0.992. All relative standard deviations were less than 10.0%. The method was employed for the determination of the analytes in C. Rhizoma. Berberine, palmatine, coptisine, epiberberine, and jatrorrhizine were present at concentrations between 9.62?±?0.87?mg/g and 0.48?±?0.05?mg/g.     

Microscopic evaluation of vesicles shed by erythrocytes at elevated temperatures

The images of human erythrocytes and vesicles were analyzed by a light microscopy system with spatial resolution of better than 90 nm. The samples were observed in an aqueous environment and required no freezing, dehydration, staining, shadowing, marking, or any other manipulation. Temperature elevation resulted in significant concentration increase of structurally transformed erythrocytes (echinocytes) and vesicles in the blood. The process of vesicle separation from spiculated erythrocytes was video recorded in real time. At a temperature of 37°C, mean vesicle concentrations and diameters were found to be 1.50 ± 0.35 × 10⁶ vesicles per microliter and 0.365 ± 0.065 μm, respectively. The vesicle concentration increased approximately threefold as the temperature increased from 37 to 40°C. It was estimated that 80% of all vesicles found in the blood are smaller than 0.4 μm. Accurate account of vesicle numbers and dimensions suggest that 86% of the lost erythrocyte material is lost not by vesiculation but by another, as yet, unknown mechanism. Microsc. Res. Tech. 76:1163–1170, 2013. © 2013 Wiley Periodicals, Inc.     

An investigation of the influences of noise on EEG power bands and visual cognitive responses for human-oriented product design

Various different human-oriented approaches are required in industrial activities. Noise is one of the most widespread sources of environmental stress. So, it is important to consider noise when we design human-oriented products. This study investigates the responses of EEG and eye movement data in order to evaluate the direct effects of low, middle, and high frequency noise on the two main physiological stress aspects: the EEG band power (alpha and beta frequency bands) and pupil response time (PRT) for a human-oriented product design. Fifteen subjects were exposed to low (100 Hz), middle (1000 Hz), and high frequency (10000 Hz) noise while awake. EEG and eye movement data were collected during noise exposure. Alpha band activity in low and high frequency noise ranges was smaller than that in no sound. Alpha band activity decreased 19.3 ± 4.5 % in the low frequency noise range. Additionally, alpha band decreased 19.5 ± 5.4% in high frequency noise range. On the other hand, Beta band activity in low and high frequency noise ranges was greater than that in no sound. Beta band activity increased 26.9 ± 7.9 % in the low frequency noise range and increased 30.6 ± 6.1% in high frequency noise range. The PRT, or visual cognitive responses, in low or high frequency noise was greater than that in no sound. PRT increased 15.3 ± 3.0% in low frequency noise range. Alternatively, PRT increased 18.1 ± 3.2% in high frequency noise range. And results of EEG and eye movement were statistically significant in low and high frequency noise (r > 0.92, p < 0.05). The findings of this study indicate that the stress induced by low frequency noise is as stressful as the stress induced by high frequency noise. Additionally, utilizing eye movement data and acquiring the PRT is useful in the analysis of human stress responses during various stressful situations in addition to the analysis of human stress responses during noise exposure.     

Effects of Er:YAG laser on bond strength of self‐etching adhesives to caries‐affected dentin

The erbium:yttrium–aluminum–garnet (Er:YAG) laser may be effective the bond strength of adhesive systems on dentine surfaces, the chemical composition and aggressiveness of adhesive systems in clinical practice. The purpose of this study was to evaluate the effects of the Er:YAG laser system with the bonding ability of two different self‐etching adhesives to caries‐affected dentine in primary molars. Ninety mid‐coronal flat dentine surfaces obtained from sound and caries‐affected human primary dentine were treated with an Er:YAG laser or a bur. The prepared surfaces were restored with an adhesive system (Xeno V; Clearfil S³) and a compomer (Dyract Extra). The restored teeth were sectioned with a low‐speed saw and 162 samples were obtained. The bond strength of the adhesive systems was tested using the micro‐tensile test method. The data were statistically analyzed. A restored tooth in each group was processed for scanning electron microscopy evaluation. The values of the highest bond strength were obtained from the Clearfil S³‐Er:YAG laser‐sound dentine group in all groups. (24.57 ± 7.27 MPa) (P > 0.05). The values of the lowest bond strength were obtained from the Xeno V‐Er:YAG laser‐sound dentine group in all groups (11.01 ± 3.89 MPa). It was determined that the Clearfil S³ increased the bond strength on the surface applied with Er:YAG laser according to the Xeno V. Microsc. Res. Tech. 77:282–288, 2014. © 2014 Wiley Periodicals, Inc.     

Shear bond strength of repairs in porcelain conditioned with laser

The aim of this study was to evaluate the shear bond strength of repairs in porcelain conditioned with laser. Sixty porcelain discs were made and six groups were formed (n = 10): G1: conditioning with laser with potency 760 mW; G2: conditioning with laser with potency 760 mW and application of 37% phosphoric acid for 15 s; G3: conditioning with laser with potency 900 mW; G4: conditioning with laser with potency 900 mW and application of 37% phosphoric acid for 15 s; G5: application of 37% phosphoric acid for 15 s (group control) and G6: application of 10% hydrofluoric acid for 2 min. The composite resin was insert of incremental layers at the porcelain surface aided with a metal matrix, and photoactivation for 20 s each increment. The specimens were submitted to a thermal cycling by 1000 cycles of 30 s in each bath with temperature between 5 and 55°C. After the thermal cycling, specimens were submitted to the shear bond strength. The results were evaluated statistically through analysis of variance and Tukey's tests with 5% significance. The averages and standard deviation founded were: G1, 11.25 (±3.10); G2, 12.32 (±2.65); G3, 14.02 (±2.38); G4, 13.44 (±2,07); G5, 9.91 (±2,18); G6, 12.74 (±2.67). The results showed that the femtosecond laser produced a shear bond strength of repairs in porcelain equal to the hydrofluoric acid and significantly superior to the use of phosphoric acid. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.     

Acute effects of splint immobilization of the forearm on in vivo microcirculation and histomorphology of the human skin

BACKGROUND: Splint immobilization of the forearm is often performed in clinical practice. Previous studies investigated the effect of immobilization on bone, cartilage, muscle, and tendon, however, the acute effects on human skin microcirculation and histomorphology remains elusive. METHODS: In 12 healthy, nonsmoking individuals (aged 29.7 ± 9.1 years) a randomly selected forearm was immobilized by splinting for 72 h, whereas the other forearm served as control. In vivo Reflectance‐Mode Confocal‐Microscopy (RMCM) was performed prior (baseline value) and postimmobilization to evaluate: quantitative blood cell flow; density of functional dermal capillaries; epidermal thickness; and granular cell size. RESULTS: At 72h forearm immobilization, quantitative blood cell flow was significantly reduced (42.86 ± 3.68 cells/min) compared to the control blood flow (53.11 ± 3.68 cells/min, P < 0.05) and dermal capillaries indicates less functional density (5.73 ± 0.63 capillaries/mm²) compared to the controls (7.04 ± 0.81 capillaries/mm², P < 0.05). Histometric assessment reveals significantly thinner epidermis following immobilization compared to the control site (40.02 ± 2.91 vs. 46.64±3.09 µm, P < 0.05). Granular cell size was significantly altered at 72 h splinting (730.1 ± 42.53 µm²) compared to the control cell size at 770.2 ± 38.21 µm². Comparison of baseline values of both forearms indicate statistically insignificance (P > 0.05) for each parameter. CONCLUSION: At 72 h splint immobilization, for the first time, significant adaptive mechanisms were evaluated on human skin microcirculation and histomorphology using in vivo RMCM. These adaptations may be considered as an incipient atrophy of the human skin. Long‐term effects of immobilization including the regenerative potential should be evaluated in further RMCM studies. Microsc. Res. Tech. 77:99–103, 2014. © 2013 Wiley Periodicals, Inc.