Ultrastructure of the myotendinous junction of the medial pterygoid muscle of adult and aged Wistar rats

The myotendon junction (MTJ) is a specialised area into the muscle fibers where the sarcoplasmic membranes connect to the collagen fibers bundles. There are few data about plasticity of the MTJ in aging processes. The aim of this study is to analyse the ultrastructure characteristics of MTJ of medial pterygoid muscle of adult and aged rats. Employing the transmission electron microscopy method, twenty male rats Wistar (Rattus norvegicus) were divided into two groups: A (n = 10) with 12 months of age; B (n = 10) 24 months of age. The animals were anaesthetised with overdose the urethane (3 g/kg, i.p.) and sacrified during the perfusion with modified Karnovsky solution. The specimens were post-fixed in a 1% osmium tetroxide solution, dehydrated in ascending concentration of ethanol and embedded in Spurr resin. The thin sections, of 90 nm thick, were counterstained with uranyl acetate and lead citrate solution, and examined in a Jeol 1010 transmission electron microscope. The fine structure of the MTJ of group A revealed the defined interdigitations and disposed in several levels of deep formations containing the collagen fibers. In the group B, such structures did not observed, detecting the projections irregular in shape, and large of extra matrix with in aspect of remodelling. In conclusion it was possible to identify the plasticity of MTJ in the group B which presented several morphological alterations comparing to the adult animals. These data of group B suggested the occurrence of aging processes in the MTJ in rats.     

The collagen type I segment long spacing (SLS) and fibrillar forms: Formation by ATP and sulphonated diazo dyes

The collagen type I segment long spacing (SLS) crystallite is a well-ordered rod-like molecular aggregate, ∼300 nm in length, which is produced in vitro under mildly acidic conditions (pH 2.5–3.5) in the presence of 1 mM ATP. The formation of the SLS crystallite amplifies the inherent linear structural features of individual collagen heterotrimers, due to the punctate linear distribution and summation of the bulkier amino acid side chains along the length of individual collagen heterotrimers. This can be correlated structurally with the 67 nm D-banded collagen fibril that is found in vivo, and formed in vitro. Although first described many years ago, the range of conditions required for ATP-induced SLS crystallite formation from acid-soluble collagen have not been explored extensively. Consequently, we have addressed biochemical parameters such as the ATP concentration, pH, speed of formation and stability so as to provide a more complete structural understanding of the SLS crystallite. Treatment of collagen type I with 1 mM ATP at neutral and higher pH (6.0–9.0) also induced the formation of D-banded fibrils. Contrary to previous studies, we have shown that the polysulphonated diazo dyes Direct red (Sirius red) and Evans blue, but not Congo red and Methyl blue, can also induce the formation of SLS-like aggregates of collagen, but under markedly different ionic conditions to those employed in the presence of ATP. Specifically, pre-formed D-banded collagen fibrils, prepared in a higher than the usual physiological NaCl concentration (e.g. 500 mM NaCl, 20 mM Tris-HCl pH7.4 or x3 PBS), readily form SLS aggregates when treated with 0.1 mM Direct red and Evans blue, but this did not occur at lower NaCl concentrations. These new data are discussed in relation to the anion (Cl⁻) and polyanion (phosphate and sulphonate) binding by the collagen heterotrimer and their likely role in collagen fibrillogenesis and SLS formation.     

Structure relates to elastic recoil and functional role in quadriceps tendon and patellar ligament

Tendons and ligaments have similar but slightly different structure and composition. Crimps of tendons and ligaments are morphological structures related to the elastic functional properties of these connective tissues. Aim of this study was to investigate the morphological arrangement of collagen fibres, fibrils and crimping pattern of suprapatellar (rectus femoris tendon-RFT and vastus intermedius tendon-VIT) and infrapatellar connective tissues (patellar ligament-PL) to relate their structural aspects to their common function role of leg extension. RFT, VIT and PL were removed from knees of Sprague–Dawley rats and light and electron microscopy (TEM and SEM) performed. Sagittal sections showed that collagen array and crimping pattern were similar in RFT and PL but differed from VIT. Morphometric analysis confirmed that crimp number was about the same in RFT and PL (5.4 ± 1.4 and 6.1 ± 2.8 respectively), but it was almost three times higher in VIT (14.5 ± 4.7). Similarly crimp top angle in RFT and PL (141.5 ± 15.0° and 146.2 ± 12.2° respectively) was significantly higher than in VIT (122.3 ± 14.8°) and the crimp base length was more than twice as wide in RFT (75.5 ± 22.6 μm) and PL (72.3 ± 28.9 μm) than in VIT (36 ± 14.1 μm). The smaller, fewer and most crimped crimps in VIT show that this tendon has a greater elastic recoil and responds to higher forces as among quadriceps muscles the vastus intermedius belly contributes the most during knee extension. By contrast, RFT acting as a “stopper” tendon also plays a ligament role by limiting an excessive flexion of the joint during postural rest position of the knee.     

Collagen fibers evaluation after rhBMP-2 insertion in critical-sized defects

The objective of this investigation was to assess the quantity of collagen fibers with different dosages of recombinant human bone morphogenetic protein, type 2 (rhBMP-2) associated with two different carriers, monoolein and poloxamer gels, in critical bone defects created in the calvaria of Wistar rats. Forty male adult Wistar rats were divided into eight groups of 5 animals each—group I: critical bone defect with application of 1 μg of rhBMP-2 combined with monoolein gel; group II: 3 μg of rhBMP-2 combined with monoolein gel; group III: 7 μg of rhBMP-2 combined with monoolein gel; group IV: 1 μg of rhBMP-2 combined with poloxamer gel; group V: 3 μg of rhBMP-2 combined with poloxamer gel; group VI: 7 μg of rhBMP-2 combined with poloxamer gel; group VII: monoolein gel only and group VIII: poloxamer gel only. A critical-sized defect of 6 mm diameter was produced in the left parietal bone using a surgical round bur and a high-speed micromotor. The bone defects were filled according to the group that animals belonged and after two weeks the rats were perfused and their calvarial bones were removed for histological processing, and collagen fibers quantification. Differences among the eight groups were statistically analyzed by Anova and Bonferroni test (p < 0.05). The results did not show statistical difference between the groups, in exception, between the comparisons II and III. According to the experimental methodology used in this research, it was observed, in a general way, a qualitative inverse relationship between collagen fibers presence and rhBMP-2 quantity inserted in the critical bone defect, associated or not to a material carrier.     

Effects of phonophoresis with Arnica montana onto acute inflammatory process in rat skeletal muscles: An experimental study

This study aimed at verifying the effects of phonophoresis associated with Arnica montana on the acute phase of an inflammatory muscle lesion. Forty Wistar male rats (300 ± 50 g), of which the Tibialis Anterior muscle was surgically lesioned, were divided into four groups (n = 10 each): control group received no treatment; the ultrasound group (US) was treated in pulsed mode with 1-MHz frequency, 0.5 W/cm² intensity (spatial and temporal average – SATA), duty cycle of 1:2 (2 ms on, 4 ms off, 50%), time of application 3 min per session, one session per day, for 3 days; the phonophoresis or ultrasound plus arnica (US+A) group was treated with arnica with the same US parameters plus arnica gel; and the arnica group (A) was submitted to massage with arnica gel, also for 3 min, once a day, for 3 days. Treatment started 24 h after the surgical lesion. On the 4th day after lesion creation, animals were sacrificed and sections of the lesioned, inflamed muscle were removed for quantitative (mononuclear and polymorphonuclear cell count) and qualitative histological analysis. Collected data from the 4 groups were statistically analyzed and the significance level set at p < 0.05. Results show higher mononuclear cell density in all three treated groups with no significant difference between them, but values were significantly different (p < 0.0001) when compared to control group’s. As to polymorphonuclear cell density, significant differences were found between control group (p = 0.0134) and US, US+A and A groups; the arnica group presented lesser density of polymorphonuclear cells when compared (p = 0.0134) to the other groups. No significant difference was found between US and US+A groups. While the massage with arnica gel proved to be an effective anti-inflammatory on acute muscle lesion in topic use, these results point to ineffectiveness of Arnica montana phonophoresis, US having seemingly checked or minimized its anti-inflammatory effect.     

Fragmentation of 370 MeV/n 20Ne and 470 MeV/n 24Mg in light targets

Total charge-changing cross sections and cross sections for the production of projectile-like fragments were determined for fragmentation reactions induced by 370 MeV/n ²⁰Ne ions in water and lucite, and 490 MeV/n ²⁴Mg ions in polyethylene, carbon and aluminum targets sandwiched with CR-39 plastic nuclear track detectors. An automated microscope system and a track-to-track matching algorithm were used to count and recognize the primary and secondary particles. The measured cross sections were then compared with published cross sections and predictions of different models. Two models and the three-dimensional Monte Carlo Particle Heavy Ion Transport Code System (PHITS) were used to calculate total charge-changing cross sections. Both models agreed within a few percent for the system ²⁴Mg + CH₂, however a deviation up to 20% was observed for the systems ²⁰Ne + H₂O and C₅H₈O₂, when using one of the models. For all the studied systems, PHITS systematically underestimated the total charge-changing cross section. It was also found that the partial fragmentation cross sections for ²⁴Mg + CH₂ measured in present and earlier works deviated up to 20% for Z = 6–11. Measured cross sections for the production of fragments (Z = 4–9) for ²⁰Ne + H₂O and C₅H₈O₂ were compared with predictions of three different semi-empirical models and JQMD which is used in the PHITS code. The calculated cross sections differed from the measured data by 10–90% depending on which fragment and charge was studied, and which model was used.     

Dielectric,ferroelectric and piezoelectric properties of (1−x)[K0.5Na0.5NbO3]−x[LiSbO3] ceramics

Lead-free (1?x)[K_0.5Na_0.5NbO₃]?x[LiSbO₃] (x=0, 0.04, 0.05 and 0.06)/(KNN-LS) ceramics were prepared by conventional solid-state reaction route (CSSR). For dense morphology pure KNN ceramic was sintered at 1120 °C and LS modified KNN ceramics were sintered at 1080 °C for 4 h, respectively. The structural study at room temperature (RT) revealed the transformation of pure orthorhombic to tetragonal structure with the increase in LS content in KNN-LS ceramics. Temperature dependent dielectric study confirmed the increase of diffuse phase transition nature with the increase in LS content in KNN-LS ceramics. The presence of orthorhombic to tetragonal (T_O?T) polymorphic phase transition temperature (PPT) ～43 °C confirmed the presence of two ferroelectric (orthorhombic and tetragonal) phases in 0.95KNN-0.05LS ceramics at RT. 0.95KNN-0.05LS ceramics showed better ferroelectric and piezoelectric properties i.e., remnant polarization (P_r)～18.7 μC/cm², coercive field (E_c)～11.8 kV/cm, piezoelectric coefficient (d₃₃)～215 pC/N, coupling coefficient (k_p)～0.415 and remnant strain ～0.07% were obtained.     

Ultrasound-intensified laccase production from Trametes versicolor

An efficient intermittent ultrasonic treatment strategy was developed to improve laccase production from Trametes versicolor mycelia cultures. The optimized strategy consisted of exposing 2-day-old mycelia cultures to 5-min ultrasonic treatments for two times with a 12-h interval at the fixed ultrasonic power and frequency (120 W, 40 kHz). After 5 days of culture, this strategy produced the highest extracellular laccase activity of 588.9 U/L among all treatments tested which was 1.8-fold greater than the control without ultrasound treatment. The ultrasonic treatment resulted in a higher pellet porosity that facilitated the mass transfer of nutrients and metabolites from the pellets to the surrounding liquid. Furthermore, the ultrasonic treatment induced the expression of the laccase gene (lcc), which correlated with a sharp increase in both extracellular and intracellular laccase activity. This is the first study to find positive effects of ultrasound on gene expression in fungal cells. These results provide a basis for understanding the stimulation of metabolite production and process intensification by ultrasonic treatment in filamentous fungal culture.     

Ultrastructural analysis of the low level laser therapy effects on the lesioned anterior tibial muscle in the Gerbil

Low level laser therapy (LLLT) is known for its positive results but studies on the biological and biomodulator characteristics of the effects produced in the skeletal muscle are still lacking. In this study the effects of two laser dosages, 5 or 10 J/cm², on the lesioned tibial muscle were compared. Gerbils previously lesioned by 100 g load impact were divided into three groups: GI (n = 5) controls, lesion non-irradiated; GII (n = 5), lesion irradiated with 5 J/cm² and GIII (n = 5), lesion irradiated with 10 J/cm², and treated for 7 consecutive days with a laser He–Ne (λ = 633 nm). After intracardiac perfusion, the muscles were dissected and reduced to small fragments, post-fixed in 1% osmium tetroxide, dehydrated in increasing alcohol concentrations, treated with propylene oxide and embedded in Spurr resin at 60 °C. Ultrafine cuts examined on a transmission electron microscope (Jeol 1010) revealed in the control GI group a large number of altered muscle fibers with degenerating mitochondria, intercellular substance containing degenerating cell fragments and budding blood capillaries with underdeveloped endothelial cells. However, groups GII and GIII showed muscle fibers with few altered myofibrils, regularly contoured mitochondria, ample intermembrane spaces and dilated mitochondrial crests. The clean intercellular substance showed numerous collagen fibers and capillaries with multiple abluminal processes, intraluminal protrusions and several pinocytic vesicles in endothelial cells. It was concluded that laser dosages of 5 or 10 J/cm² delivered by laser He–Ne (λ = 633 nm) during 7 consecutive days increase mitochondrial activity in muscular fibers, activate fibroblasts and macrophages and stimulate angiogenesis, thus suggesting effectivity of laser therapy under these experimental conditions.     

Assessing the effect of football play on knee articular cartilage using delayed gadolinium-enhanced MRI of cartilage (dGEMRIC)

The prevalence of cartilage lesions is much higher in football athletes than in the general population. Delayed gadolinium-enhanced MRI of cartilage (dGEMRIC) has been shown to quantify regional variations of glycosaminoglycan (GAG) concentrations which is an indicator of early cartilage degeneration. The goal of this study is to determine whether dGEMRIC can be used to assess the influence in cartilage GAG concentration due to college level football play. Thirteen collegiate football players with one to four years of collegiate football play experience were recruited and both knee joints were scanned using a dedicated 8-channel phased array knee coil on a 3 T MRI system. The contrast concentrations within cartilage were calculated based on the T₁ values from dGEMRIC scans. No substantial differences were found in the contrast concentrations between the pre- and post-season across all the cartilage compartments. One year collegiate football players presented an average contrast concentration at the pre-season of 0.116 ± 0.011 mM and post-season of 0.116 ± 0.011 mM. In players with multiple years of football play, contrast uptake was elevated to 0.141 ± 0.012 mM at the pre-season and 0.139 ± 0.012 mM at the post-season. The pre-season 0.023 ± 0.016 mM and post-season 0.025 ± 0.016 mM increase in contrast concentration within the group with multiple years of experience presented with a > 20% increase in contrast uptake. This may indicate the gradual, cumulative damage of football play to the articular cartilage over years, even though the effect may not be noticeable after a season of play. Playing collegiate football for a longer period of time may lead to cartilage microstructural alterations, which may be linked to early knee cartilage degeneration.     

In situ magneto-optical Kerr effect study of uncovered Fe films on ZnSe(0 0 1)

The magnetic properties of uncovered Fe/ZnSe/GaAs(1 0 0) ultrathin films have been determined in situ by magneto-optical Kerr effect (MOKE). Fe films up to 10 monolayers (ML) thick were deposited on c(2×2) Zn-rich ZnSe/GaAs(0 0 1) surfaces at 180 °C. We have studied the thickness dependence of the in-plane lattice parameter of the Fe films and of the MOKE hysteresis loops in the longitudinal geometry, at 150 K, under magnetic fields up to 0.1 T applied along the [1 1 0] and [1-1 0] directions of the ZnSe(0 0 1). Reflection high energy electron diffraction show that in the low thickness regime the Fe films present an in-plane structural anisotropy characterized by an expansion along the [1 1 0] direction. Hysteretic loops were obtained only starting from ∼5 ML Fe. We found the onset of an uniaxial magnetic anisotropy with [1 1 0] magnetic easy axis at 7 ML Fe.     

Thermoluminescence,infrared reflectivity and electron paramagnetic resonance properties of hemimorphite

Silicate mineral hemimorphite has been investigated concerning its TL, IR and EPR properties. A broad TL peak around 180 °C and a weaker and narrower peak around 360 °C were found in a sample annealed at 600 °C for 1 h and then irradiated. The deconvolution using the CGCD method revealed peaks around 132, 169, 222 and 367 °C. The reflectivity measurements showed several bands in the NIR region due to H₂O, OH and Al–OH complexes. No band was observed in the visible region. The thermal treatments were carried out from ∼110 to 940 °C and dehydration was observed, first causing a diminishing optical absorption in general and the disappearance of water and hydroxyl absorption bands. The EPR spectrum of natural hemimorphite, presented Cu²⁺ signals at g = 2.4 and g = 2.1 plus E₁′ signal superposed to Fe³⁺ signal around g = 2.0.     

Electrical and structural investigations of perovskite structure reactively sputter deposited coatings

Sr(Zr_0.84Y_0.16)_0.91O_3 ? δ (SZY) and Ba(Zr_0.84Y_0.16)_0.96O_3 ? δ (BZY) protonic conductor coatings were co-sputter deposited from metallic targets in argon–oxygen reactive gas mixtures. The chemical and structural features were investigated by energy dispersive X-ray spectroscopy and X-ray diffraction, and their morphology was assessed by scanning electron microscopy of the surface and of brittle fracture cross sections. The electrical properties of the coating were determined by complex impedance spectroscopy as a function of temperature in air. Relationships are established between the electrical properties and the morphology of the coatings. The SZY as deposited coatings is amorphous and crystallises under the convenient perovskite structure after annealing treatment at 873 K under air. The BZY as deposited coatings is crystallised at 523 K in situ under perovskite structure and a further annealing treatment increases the grain size. Conductivities and activation energies of crystallised coatings were 3.1 · 10^? 5 S cm^? 1/2 · 10^? 5 S cm^? 1 and 0.65 eV/0.71 eV after stabilization at 773 K for strontium and barium zirconate, respectively.     

A 48% Ni–Fe alloy of low coercivity and improved corrosion resistance in a cyclic damp heat test

Electric arc 30t melts, deoxidised with Mn and Si, are compared to melts additionally deoxidised with cerium. In a cyclic damp heat test the melts with Ce show better corrosion behaviour. μ₄ (DC) and H_c of samples of 1.0 mm thickness after heat treatment at 1080°C vary in the same range of values for both categories of melts. After heat treatment at 1080°C or 1180°C μ₄ (50 Hz) and H_c of samples of 0.2 mm thickness are impaired by 20% or more for melts with Ce. For melts with Ce the strong binding of the sulphur to the deoxidiser impedes desulphurization. With small strip thicknesses, the strong binding energy of Ce to oxygen causes additional internal oxidation during heat treatment . Both deteriorate the magnetic properties. At the larger thickness of 1.0 mm there is no noticeable internal oxidation and/or desulphurization in both categories of melt due to the strip thickness.     

Magnetic fluid thermochemotherapy of murine tumors

Four series of water-based dextran-ferrite (DF) magnetic fluids (MFs) containing Melphalan (MP) were prepared. Their saturation magnetization (M_s) was from 0.8 to 7.8 kA/m; specific power absorption rates were 240 W/g Fe. After nine courses of magnetic fluid thermochemotherapy with MP containing DF MF (M_s 7.5 kA/m, pH 6.6, ζ+15 mV) at 44–46 °C for 30 min in an AC magnetic field (0.88 MHz, 9.3 kA/m, 0.3 kW), complete P388 tumor regression was seen in 30% of BDF₁ mice. Furthermore, a life span increase of 180% was achieved.     

Studies of structural and electrical properties on four-layers Aurivillius phase BaBi4Ti4O15

BaBi₄Ti₄O₁₅ (BBT) ceramic was synthesized using mixed oxide route and the structural and electrical properties were investigated systematically. The structural studies confirmed it to be an n=4 member of the Aurivillius oxide. A broad dielectric peak with frequency dependent dielectric maximum temperature was observed. The dielectric relaxation obeyed the Vogel–Fulcher relation wherein f₀=8.37E+14 Hz, E_a=0.13 eV, and T_f=608.18 K. The diffuseness parameter γ established the relaxor nature and it was attributed to the A-site cationic disorder. The specimen exhibited the excellent reproducibility in the measurements of displacement current, a remnant polarization of 5.4 μC/cm², and a coercive field of 4.03 MV/m. The room temperature piezoelectric coefficient d₃₃ was found to be 23 pC/N and the field-induced strain S was about 0.018% at the 8 MV/m electric field.     

Analysis of temperature-dependent electrical resistivity of ZnO nano-structures

The temperature–dependent electrical resistivity ρ(T) in metallic and semiconducting phase of ZnO nanostructures is theoretically analysed. ρ(T) shows semiconducting phase in low temperature regime (140 K<T<180 K), shows an absolute minimum near 180 K and increases linearly with T at high temperatures (200 K<T<300 K). The resistivity in metallic phase is estimated within the framework of electron–phonon and electron–electron scattering mechanism. The contributions to the resistivity by inherent acoustic phonons (ρ_ac) as well as high frequency optical phonons (ρ_op) were estimated using Bloch–Gruneisen (BG) model of resistivity. The electron–electron contributions ρ_e?e=BT² in addition with electron–phonon scattering is also estimated for complete understanding of resistivity in metallic phase. Estimated contribution to resistivity by considering both phonons, i.e., ω_ac and ω_op and the zero limited resistivity are added with electron–electron interaction ρ_e–e to obtain the total resistivity. Resistivity in Semiconducting phase is discussed with small polaron conduction (SPC) model. The SPC model consistently retraces the low temperature resistivity behaviour (140 K<T<180 K). Finally the theoretically calculated resistivity is compared with experimental data which appears favourable with the present analysis in wide temperature range.     

Study of the microstructure and mechanical properties of white clam shell

The microstructure and mechanical properties of white clam shell were investigated, respectively. It can be divided into horny layer, prismatic layer and nacreous layer. Crossed-lamellar structure was the microstructural characteristic. The extension direction of lamellae in prismatic layer was different from that in nacreous layer, which formed an angle on the interface between prismatic layer and nacreous layer. The phase component of three layers was CaCO₃ with crystallization morphology of aragonite, which confirmed the crossed-lamellar structural characteristic. White calm shell exhibited perfect mechanical properties. The microhardness values of three layers were 273 HV, 240 HV and 300 HV, respectively. The average values of flexure and compression strength were 110.2 MPa and 80.1 MPa, respectively. The macroscopical cracks crossed the lamellae and finally terminated within the length range of about 80 μm. It was the microstructure characteristics, the angle on the interface between prismatic and nacreous layer and the hardness diversity among the different layers that enhanced mechanical properties of white calm shell.     

Morphological and ultrastructural examination of senescence in Morchella elata

In recent years, the artificial cultivation of Morchella mushrooms that belong to Elata Clade, including Morchella elata, has been developed rapidly in China. However, the prominent problem of spawn aging has been frustrating the morel farming. In this paper, aging in M. elata was achieved from 12 to 17 subcultures and lifespan of 1536–2256 h by successive subculturing. The lifespan can be roughly divided into juvenile phase and senescent phase with respect to the mycelia linear growth rate. After a certain period of rapid growth with almost constant rate at the juvenile phase, the isolate entered the senescent phase characterized by slow down of mycelia growth, producing pigments ahead of time and final death of the apical hyphae. The period of the senescent phase was definitely 240–288 h; while that of the juvenile phase was diverse relying on different isolates. Moreover, microscopic study showed that angles between the leading and primary hyphae increased constantly with aging. In senesced hyphal cells of M. elata, the typical characteristics of autophagy (enlargement of vacuoles and existence of organelles sequestrated lysosomes) and apoptosis (condensation of the cytoplasm and nuclear and plasmolysis) were observed. In addition, in the final stage of senescence, the apical hyphae collapsed with the plasma membrane and all the cellular organelles disrupted, indicated a necrotic mode of cell death. Taken together, these data revealed the involvement of autophagy, apoptosis and necrosis in senescence of M. elata. The characterization and molecular mechanism of autophagy, apoptosis and necrosis need further study and the systematic study of morel aging will be beneficial for the healthy development of morel farming.