Physiologic combinations of pH, lactate, andATP activate DRG neurons and produce pain in humans [11,12]. In the present study, we extend these findings by showing this low dose com- bination of (pH 7.5, 15 μM lactate, 76 nM ATP) produces musclehyperalgesiaand is synergis- tic. It is curious that the lowest concentration combination produced mechanical hyperalgesia. This may mean that increasing concentration of these compounds is not sufficient to produce hyperalgesia; rather, concentrations must be within a specific range for the receptors to be acti- vated. In subsequent experiments, we used a higher dose oflactate with physiological doses ofATPand pH (pH 6.0, 10 mM lactate, 2.4 μM ATP), doses similar to that used in humans . We show the interaction between these 3 substances is synergistic, and that their effects are long-lasting. Lower concentrations of these compounds (pH 7.3, 400 nM ATP, 5 mM lactate) injected into the muscle produce warmth and fatigue sensations, while pain is reported with injection of higher concentrations of the combined compounds in humans (pH 7.2, 500 nM ATP, 10 mM lactate—pH 6.6, 5 μM ATP, 50 mM lactate) . The fact that three ineffective doses when combined together cause significant decreases inmuscle withdrawal threshold sug- gests protons, lactate, andATP act synergistically to produce mechanical hyperalgesia [20,21]. Further, we show combining all 3 substances is necessary to produce the mechanical hyperalge- sia, as each paired combination failed to produce mechanical hyperalgesia. This is consistent with previous studies showing acid-evoked currents and calcium influx inmuscle DRG are potentiated, and the greatest effects occur, by combining all 3 metabolites [11,13,14]. The present behavior studies also show a slow onset requiring 1–2 hours for maximal hyperalgesia. This hyperalgesia lasts for hours after a single injection, suggesting activation of cellular pro- cesses which are independent of ion channel effects, activation of other cell types such as mac- rophages , and/or triggering release of inflammatory cytokines .
After the immobilization, with the musclein the short position, the muscular atrophy and the proliferation of the connective tissue are considered the most important muscular adaptations. Both alterations are caused by disequilibrium between the synthesis and degradation of the muscular and collagen proteins (10). Studies show that the normal collagen remodeling is of only 1.3% a day in muscles ofrats, while the synthesis rate of muscular proteins is 11.8%. This way, during the immobilization, a higher loss of muscular proteins occurs with a proportional increase in the disorganized intramuscular connective tissue (ICT), reducing the flexibility of the muscular tissue and the cross-section area (CSA) of the muscular fibers (11,12), resulting in a smaller functional capability. On the other hand, the muscular stretching exercise is known as a powerful stimulus to smooth the atrophy and to induce the muscular hypertrophy (4,5,13,14). A recent study showed that stretch bouts applied to the soleus muscleof a rat 3 times a week, during 40 minutes, increased the number of serial sarcomeres and the cross-section area (4). Other studies have also identified the beneficial effectof the muscular stretching for the remodeling and reorganization of collagen fibers of the ICT (1,14).
Experimental Design and Drug administration All the animals of either sex were divided into two groups of six male and six female and were treated with tolfenamic acid alone (4 mg/kg, intramuscular) and tolfenamic acid along with moxifloxacin (5 mg/kg, intramuscular), respectively. Tolfenamic acid was formulated in 30 % labrasol solution in phosphate buffer saline (pH 7.2) (vehicle-I), where as the moxifloxacin was formulated for dose administration in 5% Twin-80, 5% DMSO (dimethyl sulphoxide) in Milli-Q water (w/v) (vehicle-II). Doses were calculated according to body weight of the animals and administrated as per concentration strength of formulation. The drugs were administered by intramuscular route using sterile 1 ml syringe and needle (26 G, 0.45mm x 13mm) in deep gluteal muscle.
The system of supply chain of tuna commodity can be said that it is very short. This relates to the high price of fresh tuna and the requirements of fresh fish quality determined by buyers in export market. The figure 3 shows that the whole tuna fish from the fishermen that belongs to grade A (according to the PPN fishermen in Pelabuhan Ratu) are sold to the big fishermen. Big fishermen are fishery companies that have big armada for tuna fishing and logistic armada to supply the need of the tuna armada and tuna traders. The big fishermen have double functions – selling the fish caught alone and storing the tuna to other fishermen and supplying the need of tuna armada. Fish with grades A, B+, B- and C exported to America, Amsterdam, Spanyol and Asean continent must be processed first into loin, saku saku, steak and fillet. Other remaining fish are called tetelan fish from the remaining fish with CO gas and fish reject. This type of Tuna fish is sold with various prices. The market chain of this market fish in Pelabuhanratu is relatively short, the role of industry is very strong in which big companies invest their modal and manage the system from downstream to upstream. The big companies have big armada by themselves. The benefit of the business is mostly enjoyed by the big industries, and local businessmen only enjoy small part of the benefit. Fish from fishermen in Pelabuhanratu are brought to the fish auction to sell. Fish from outside the area are like long tail tuna (tongkol), skipjack tuna and others. Usually the fish are directly sold to the big traders in Pelabuhan Ratu. Fish accepted by the big traders are then bought by the retailers and processors as well. The fish are then sliced into pindang and bought by the fish manufacturers/ pemindang (Figure 4). The analysis of VA of long tail tuna (tongkol) and skipjack tuna commodity consists of two chains, i.e. VCA of long tail tuna (tongkol) from the retailers and manufacturers. The explanation below will discuss the retailers and manufacturers. The development of skipjack tuna and long tail tuna (tongkol) as industrial products in Pelabuhan Ratu depends on the supply outside Pelabuhan Ratu, especially from Muara Baru.
To investigate whether the high Venus/CFP ratios of the dot- like structures detected in cells replicating SGR-ATeam are located at the HCV RC, FRET images of SGR-AT1.03- replicating cells were analyzed, followed immunofluorescence analysis of cells fixed and stained with either anti-NS5A or anti- NS3 antibodies (Figure 5C). Confocal fluorescence microscopy at high magnification demonstrated that the high Venus/CFP ratios that were identified in foci of various sizes were co-localized with NS5A and NS3 that were possibly membrane-bound within the cytoplasm of the viral replicating cells. Some of the NS3- or NS5A- labeled proteins that were identified by immunofluorescence were not associated with high Venus/CFP ratios. These results are consistent with previous reports, which demonstrated that only Figure 4. Development of NS5A-ATeam and SGR-ATeam to enable real-time monitoring ofATP. (A) Schematic representation of the ATeam and NS5A-ATeam used in this study. ATeam genes were inserted into the 39 region of a HA-NS5A expression vector to generate NS5A-ATeam. The underlined sequences indicate NS5A residues. The insertion site was between residues 2394 and 2395, numbered according to the polyprotein of the HCV JFH-1 isolate. CMV, Cytomegalovirus promoter; CAG, CAG promoter; ATP b.p, ATP binding protein. HA, HA tag. (B) Huh-7 cells were transfected with ATeam and NS5A-ATeam constructs. Forty-eight hours post-transfection, the Venus/CFP ratios of each cell were calculated from fluorescent images acquired with a confocal microscope in the same way as described in the legends for Figure 2. Each plot shows the ratio of individual cells. Horizontal lines represent means. (C) Schematic representation of the SGR and SGR-ATeam plasmids used, with or without the firefly luciferase gene (Fluc). HCV polyproteins are indicated by the open boxes. ATeam genes were inserted into the same site in the NS5A C-terminal region. Bold lines indicate the HCV UTR. EMCV IRES is denoted by the gray bars. Pol I P, Pol I promotor; dC, 59 region of Core gene; Pol I T, Pol I terminator. (D) Replication levels of SGR/luc-AT1.03 in transfected cells were determined by luciferase assay 1–5 days post-transfection. SGR/luc and SGR/luc-GND were used as positive and negative controls, respectively. Values given were normalized for transfection efficiency with luciferase activity determined 24 h post-transfection. All data are presented as means and SD for three independent samples. (E) Huh-7 cells were transfected with constructs encoding NS5A, NS5A-AT1.03, SGR, SGR-AT1.03, SGR/luc or SGR/luc-AT1.03, followed by immunoblotting with anti-NS5B or anti-beta- actin antibody. (F) Cells transfected with constructs encoding NS5A, NS5A-AT1.03, SGR or SGR-AT1.03 were analyzed by immunoblotting with anti- NS5A, anti-NS5B or anti-beta-actin antibodies.
To further investigate the mechanism by which nicotine acting through the nAChR-Na,K-ATPase complex regulates Na,K- ATPase activity, we examined the relationship between PKC activation, PLM phosphorylation and [ 3 H]ouabain (2 m M) binding, which reflects a2 Na,K-ATPase content in the sarcolem- ma (Fig. 6). Acute activation of PKC by PMA (100 nM) increases PLM phosphorylation at Ser 63 and Ser 68 (Fig. 6A), similar to the effectof chronic nicotine. PKC activation and PLM phosphory- lation follow a parallel time course; both are stimulated within 30 minutes and the changes are sustained for up to 120 min. The same treatment (100 nM PMA) applied to intact rat skeletal muscle does not change the content of a2 Na,K-ATPase in the plasma membrane (Fig. 6B). Over the same time period, ouabain binding to the Na,K-ATPase reached equilibrium and there was no difference in the maximum amount of ouabain bound between control and PMA treated. [ 3 H]ouabain binding to intact skeletal muscle measures only Na,K-ATPase pumps in the plasma membrane which have the extracellular ouabain binding site accessible to ligand. This result suggests that acutely activated PKC stimulates Na,K-ATPase specific activity by a mechanism which includes PLM phosphorylation, without change in the total content of Na,K-ATPase in the plasma membrane.
During cooling, the crystallisation of cast iron deviates from the equilibrium state. This means that the austenite gets supersaturated with carbon and chromium, which greatly reduces the temperature of its transformation. Under these conditions, it is possible to obtain at room temperature the matrix of austenitic or austenitic-pearlitic character, as shown by respective microstructures. Total content of the carbide phase can be determined from F. Maratray’s  formula, namely:
mechanical grinding and polishing. Back scattered electrons (BSE) were utilized in SEM in order to reveal difference in chemical compositions of microcomponents present in particular samples. The SEM investigations were used to reveal the distribution of graphite and other big particles. Transmission electron microscopy (TEM), on the other hand, was applied for examination of nanosized secondary precipitates, i.e. vanadium or niobium carbides and/or nitrides (or carbonitrides). The thin foil technique was implemented for this purpose. The 3 mm disks were ground down on sand papers and then dimpled to about 0.1 mm thickness. Afterwards the disks were further thinned in an ion mill until a perforation had appeared. The TEM investigation was carried out by means of a JEOL 2010 ARP analytical scanning transmission electron microscope operating at acceleration voltage of 200 kV. Imaging was performed by conventional transmission mode while for chemical analysis (X-ray Energy Dispersive Spectroscopy - EDS) the nanoprobe mode was utilized. The nanoprobe mode enabled to obtain electron probes approaching a few nanometers in diameter (practically about 10 nm because at smaller electron probes the number of X-ray counts is usually too low for analysis). The EDS analysis was performed by Oxford-Link system attached to the microscope. The Oxford- Link system was equipped with Si(Li) detector. This system detects all elements down to boron. In order to examine the crystallography of precipitates the Selected Area Diffraction (SAD) patterns analysis was also performed.
Goats are widely distributed around the world and have been source of human nutrition. Goat meat is without a doubt one of the staple red meats in human diets (Webb et al., 2005). h ere is some evidence for improved bioavailability of organic compared to inorganic trace mineral source (Spears, 1996). According to Bekhit et al. (2005) and Koohmaraie (1990) Zn could inl uence meat quality (for example colour and texture). h e present study was therefore designed to compare the ef ect of organic andin- organic zinc forms on meat quality and concentration inmuscle (m. triceps brachii) of young goats.
Table 2 gives the results of mechanical tests carried out on the low-alloy cast steel with additions of vanadium and compares them with the results of previous studies made on this cast steel (designated as P1 in Tables 1 and 2) subjected to heat treatment recommended by the respective standard .
Objective: to analyze the morphometric and mechanical characteristics of the soleus and gastrocnemius muscles after immobilization in a shortened position. Methods: 20 Wistar rats (250 ± 20g) were divided equally into immobilized and control groups. The left hind limb was immobilized by means of an acrylic resin orthosis, with the ankle joint at maximum plantar flexion. After seven days of immobilization, the muscle mass, number and length of sarcomeres in series, muscle fiber cross-sectional area, density of the intramuscular connective tissue area and tensile strength of the triceps surae muscle were evaluated. The data were analyzed by the ANOVA and Tukey tests (p< 0.05). Results: The immobilized soleus muscle presented changes in all the morphometric variables analyzed, while some of these changes were not observed in the gastrocnemius muscle. Analysis of the mechanical test showed that the immobilized group presented a 20% decrease in the maximum tensile muscle strength. Conclusion: The results from this study showed that short-term immobilization causes changes to the morphometric parameters of the muscle fibers, with repercussions onmuscle mechanics. These results suggest the need for rehabilitation of muscles subjected to immobilization, even if only for a short period, in order to achieve early recovery of normal muscle characteristics.
were kept in a termal cushion in order to prevent hypothermia. Heart rate, invasive blood pressure and rectal temperature were continuously monitored in all studied animals. The right cervical and median abdominal area were then iniltrated with 2mg/kg of bupivacaine 0.125%. A right cervical incision was performed and the right jugular vein and the right carotid artery were cannulated under direct vision using a 24 G angiocath. The right jugular vein was cannulated to infuse the study solution and the right carotid artery to establish an invasive blood pressure monitoring and to obtain blood samples. After establishment of the central venous access , the luid replacement solution was started in both groups using an infusion pump( ANNE, Abbott Laboratories , Abbott Park, USA) using 5ml.kg -1 hr -1 Ringer’ lactate (HalexIstar, Sao
It has been stated that the surface layer produced in cast steel was of a composite character, as confirmed by the presence of TiC carbides obtained in SHS synthesis. The thickness of this layer ranged from 550 to 1200µm (Fig. 3-4) and depended on the amount of substrates of the exothermic reaction applied on mould walls. The size of the TiC carbides produced by synthesis ranged from 0,5µm to 4µm, andin the case of clusters of the coagulated carbides, phases of even 20µm were observed (Fig. 8). Deep etching and fracture of the composite layer of the casting revealed a non-typical, spheroidal shape of the TiC particles (Fig. 9-10). Considering the conditions of alloy solidification and the low value of the heat transfer coefficient of a ceramic mould, it can be concluded that the crystals of titanium carbide were growing under the conditions of insufficient undercooling, hindering the formation of faceted crystals, typical of TiC. This effect can be the result of a local quasi thermodynamic equilibrium, which makes crystals growing in the liquid coagulate in an attempt to reach the minimum surface energy.
in body weight are frequently observed in the state of diabetes. In the present study, induction of diabetes by alloxan also produced a decrease in body weight. The cinnamon administration showed a significant increase of weight compared to diabetic control rats. Decrease in body weight of diabetic rats was possible due to catabolism of fats and protein, even though the food intake was more in diabetic rats than control. Due to insulin deficiency, protein content was decreased in muscular tissue by proteolysis (Vats, Yadav, Grover, 2004). Similar results were obtained by Mahmood et al. (2011) and El Desoky et al. (2012) who reported a decrease of weight in alloxan diabetic rats. As shown in this study, cinnamon was an effective agent contributing to the reduction of glucose levels in serum. The high serum glucose level induced by alloxan was decreased by the cinnamon powder supplementation for 50.02% during the fourth week of treatment (p < 0.001). Our results corroborated the findings of Mahmood et al. (2011) about the elevation of blood glucose level in alloxan-induced diabetic rats, and a reduction in this level after cinnamon extract administration. Rekha, Balaji and Deecaraman (2010) reported that the increased levels of plasma glucose in streptozotocin-induced diabetic rats were lowered by the administration of cinnamon extract. Mahera (2010) and Ping, Zhang and Ren (2010) reported that oral
Studies have shown elevated intraneuronal NGF levels during inlammatory processes. hese increased levels are associated with increased BDNF expression, which, although not contributing to the processing of nociceptive informa- tion in normal circumstances, contributes to inlammatory hypersensitivity 30 . It has been reported that the use of a re-
lyzed immediately (model 23L, Yellow Springs Instruments) in the sample from one of the vacutainers (lithium heparin). Blood was then centrifuged and the plasma was kept on ice for later analysis. Plasma electro- lyte concentrations were measured using ion selective electrodes (Kodak Ektachem Ana- lyzer 700XR), protein was measured by re- fractometry, and osmolality was measured by freezing point depression (model 3MO). The blood sample collected into EDTA vacutainers was used for hematocrit and he- moglobin determination (Coulter-Counter, Model S-plus STKR; Miami, FL, USA).
Aerobic glycolysis is a seemingly wasteful mode ofATP production that is seen both in rapidly proliferating mammalian cells and highly active contracting muscles, but whether there is a common origin for its presence in these widely different systems is unknown. To study this issue, here we develop a model of human central metabolism that incorporates a solvent capacity constraint of metabolic enzymes and mitochondria, accounting for their occupied volume densities, while assuming glucose and/or fatty acid utilization. The model demonstrates that activation of aerobic glycolysis is favored above a threshold metabolic rate in both rapidly proliferating cells and heavily contracting muscles, because it provides higher ATP yield per volume density than mitochondrial oxidative phosphorylation. In the case ofmuscle physiology, the model also predicts that before the lactate switch, fatty acid oxidation increases, reaches a maximum, and then decreases to zero with concomitant increase in glucose utilization, in agreement with the empirical evidence. These results are further corroborated by a larger scale model, including biosynthesis of major cell biomass components. The larger scale model also predicts that in proliferating cells the lactate switch is accompanied by activation of glutaminolysis, another distinctive feature of the Warburg effect. In conclusion, intracellular molecular crowding is a fundamental constraint for cell metabolism in both rapidly proliferating- and non-proliferating cells with high metabolic demand. Addition of this constraint to metabolic flux balance models can explain several observations of mammalian cell metabolism under steady state conditions.
ent model, the writhing test in mice, zymosan, or acetic acid-induced writhing was also mediated by cyclooxygenase products and sympathomimetic amines, the release of which was mediated by TNF-␣, IL-1␤, and IL-8 (Duarte et al., 1988; Thomazzi et al., 1997). These cytokines appear to be released by resident peritoneal macrophages and mast cells since the depletion of these cells from the mouse peritoneal cavity abolished the acetic acid- or zymosan-induced writhing re- sponse. Furthermore, the increase in the numbers of these cells in the peritoneal cavity enhanced the number of writh- ing movements induced by both stimuli (Ribeiro et al., 2000). Within the last decade, cytokines generally regarded as anti-inflammatory have been described that inhibit the pro- duction of other cytokines such as IL-1␤, -6, -8, and TNF-␣, which are generally regarded as proinflammatory. The class of anti-inflammatory cytokines includes IL-4, -10, -13, and transforming growth factor-␤ (Hart et al., 1989; Fiorentino et al., 1991; Cassatella et al., 1993; Callard et al., 1996).
Objetivo: Analisar o efeito da proteína sericina associada ao exercício físico de natação na histomorfometria do músculo plantar de ratos Wistar. Métodos: Foram utilizados 40 ratos adultos divididos aleatoriamente em 5 grupos, com 8 animais cada: Controle, Lesão, Sericina, Natação, Natação e Sericina. Três dias após a compressão do nervo isquiático, os Grupos Natação e Natação e Sericina foram submetidos ao exercício físico de natação durante 21 dias. Após, os animais foram sacrificados, e o músculo plantar foi processado. Resultados: Não houve diferença da área da secção transversa entre os grupos, quantidade de núcleos periféricos, quantidade de fibra, relação núcleo/fibra e diâmetro menor. A análise morfológica revelou que no Grupo Exercício ocorreu hipertrofia das fibras, assim como nos Grupos Exercício e Sericina e Lesão, o dano muscular foi evidente. O percentual de conjuntivo intramuscular parece ter sido mantido no Grupo Exercício em relação aos demais grupos. Conclusão: A associação da proteína sericina e exercício físico de natação não foi eficiente na melhora das propriedades musculares, embora a aplicação do exercício físico tenha sido eficiente na manutenção do conjuntivo intramuscular, e no não agravamento dos efeitos deletérios consequentes da lesão nervosa periférica.