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Acute endurance exercise has been shown to modulate cyclooxygenase (COX)-2 expression, which is suggested to affect neuronal plasticity and learning. Here, we investigated the effect of regular strength and endurance training on cerebral COX-2 expression, inflammatory markers in the brain, and circulating cytokines. Male C57BL/6N mice were assigned to either a sedentary control group (CG), an endurance training group (EG; treadmill running for 30 min/day, 5 times/wk, 10 wk), or a strength training group (SG; strength training by isometric holding, same duration as EG). Four days after the last bout of exercise, blood and brain were collected and analyzed using real-time PCR, Western blot, and a multiplexed immunoassay. In EG, COX-2 mRNA expression in the cortex/hippocampus increased compared with CG. A significant increase of COX-2 protein levels was observed in both cortex/hippocampus and hypothalamus of mice from the SG. Nuclear factor (NF)κB protein levels were significantly increased in mice from both exercise groups (hypothalamus). A significant increase in the expression of microsomal prostaglandin E synthase (mPGES), an enzyme downstream of COX-2, was found in the hypothalamus of both the EG and SG. While most inflammatory factors, like IL-1α, IL-18, and IL-2, decreased after training, a positive association was found between COX-2 mRNA expression (cortex/hippocampus) and plasma IL-6 in the EG. Taken together, this study demonstrates that both endurance as well as strength training induces COX-2 expression in the cortex/hippocampus and hypothalamus of mice. A potential mediator of COX-2 expression after training might be circulating interleukin (IL)-6. However, further research is necessary to elucidate the role of inflammatory pathways on brain plasticity after training.
- resistance exercise
- prostaglandins
- cytokines
- cyclooxygenase-2
- microsomal PGE synthase
Both strength as well as endurance training induce COX-2 expression in the brain. The induction of cyclooxygenase (COX)-2 expression in the cortex/hippocampus and hypothalamus appears to be more pronounced following strength training. In the hypothalamus, strength training affected the COX-2 pathway including inflammatory signaling via nuclear factor (NF)κB. These signals seem to be independent from systemic inflammatory processes. A potential mediator of COX-2 expression after training might be the systemically released interleukin (IL)-6.
acute and regular exercise training has been shown to modulate memory formation, in particular neuronal plasticity such as synaptogenesis and neurogenesis (39, 42). However, the type of exercise training necessary to ensure these beneficial effects on memory is less clear. While some studies have shown that an increase in cardiovascular fitness is mandatory to achieve (5), others have found that increases in physical activity, by doing strength training, without an increase in cardiovascular fitness also can have beneficial cognitive effects (10). Strength training is accompanied by lower oxygen demands and cardiorespiratory drive than is endurance training and focuses on exercising specific muscle groups. During and after strength training, growth factors and cytokines are released, which might play a role in beneficial regeneration and adaptation processes (11). Moreover, strength training has been shown to be effective in the prevention and treatment of many metabolic and cardiovascular diseases (3). Results obtained from a recent study suggest that a long period of strength training can increase systemic levels of neurotrophic factors, which is accompanied by an improvement of cognitive performance (31). However, no studies have investigated the potential signaling pathways involved in improving cognitive function following strength training.
In general, results obtained from animal studies appear to indicate that an exercise-induced increase in neurotrophic factors, crucial for synaptic efficacy, neuronal connectivity, and use-dependent plasticity, mediate the beneficial effects of exercise (10). In particular, increased levels of brain-derived neurotrophic factor (BDNF) are assumed to induce neurotrophic downstream pathways in different brain regions like the cortex or the hippocampus following exercise training (4). Moreover, it was recently shown that endurance exercise affected cyclooxygenase (COX)-dependent pathways in the brain. COX is widely distributed in central nervous system neurons and thus enables the synthesis of prostaglandins (PGs) from arachidonic acid in the brain. In particular, COX-2 is upregulated in response to external stress signals and inflammatory cytokines (37). Besides mediating inflammatory responses in the brain, COX-2 and microsomal PGE2-1 synthase (mPGES) also play a role in memory formation (14, 40). In this regard, it was shown that COX inhibitors like ibuprofen inhibit learning-induced increases in PG production. Referring to exercise, it was shown that BDNF and COX interact during intercellular signaling cascades associated with synaptic plasticity (40). However, the activation of most of these signaling pathways has been demonstrated for an acute single bout of exercise; how they respond to long-term exercise training or specifically strength training has not been investigated to date. The term "regular exercise training or chronic exercise" describes the process of a repeated series of acute bouts of exercise and regeneration, which leads to a subsequent improvement in baseline physical fitness by inducing functional and structural adaptations (8).
Therefore the aim of the current study was to investigate the effect of a regular strength and endurance training program on cerebral COX-2 expression, inflammatory pathways in the brain, and cerebral cytokines. To identify potential systemic mediators of COX expression, several plasma cytokines were analyzed. We have shown that long-term endurance and resistance training induce COX-2-related pathways independent of peripheral inflammation. Thus the positive effects of this kind of exercise on learning and memory that have previously been suggested might be related to specific changes in the expression of enzymes in the induction of protaglandins.
Animals and experimental groups.
Eighteen male C57BL/6N mice (aged 10–12 wk) were randomly assigned to an endurance training group (EG), a strength training group (SG), and a control group (CG), which remained sedentary and served as age-matched controls. The initial body weights can be found in Table 1. Four to six mice per cage were housed at 21 ± 1°C in standard cages with free access to food and tap water. The protocol was approved by the Animal Welfare Officer of the Justus-Liebig-University and the Regierungspräsidium Giessen (no. 94/2010).
Exercise training protocols.
All animals were housed on a reverse light-dark cycle (lighting on from 2100 to 0900), and exercise training and testing were conducted during the dark cycle. Strength training was undertaken according to the experimental setup and protocol previously described by us (16). Briefly, mice gripped horizontal wires of a metal mesh, placed in a vertical position, with their front and hind paws. The orientation of the mice was such that their head was held upright. A plate placed surrounding the mesh prevented movement away from it (Fig. 1). Strength training was performed five times per week for 3 min and three series for 10 wk. The break between each series was 1 min. Endurance training consisted of mice running on a treadmill for 30 min/day, five times per week for 10 wk. Running speed was 0.26 ± 0.05 m/s, which corresponded to mice running at about 80% of their V̇o2 max. The control group (CG) did not perform any strength or endurance training. The time span between the last training session and tissue preparation was ∼4 days to prevent any confounding effects of an acute bout of exercise.
Endurance and strength testing.
Isometric strength was analyzed by measuring maximum holding time (MHT) in the same experimental setup described in the strength training protocol (15). Briefly, the time taken for mice to release both back paws from the wire was recorded. Endurance capacity was determined by using a treadmill spirometry (custom-made). After 10 min of acclimatization in the treadmill chamber, mice performed a continuous, progressive exercise test until exhaustion to measure V̇o2 max. The test started at 0.15 m/s, and the speed was increased by 0.05 m/s every 3 min.
Blood samples and tissue collection.
After the 10-wk training period and at least 4 days after the last training session, mice from the three groups were euthanized, and cardiac blood was collected immediately after killing by using a heparinized syringe. Brains of each animal were carefully excised, weighed, and quickly frozen in liquid nitrogen. Frozen brains were cut using a cryostat (2800 Grigocut E; Reichert-Jung, Nußloch, Germany). Approximately 22 × 80-μm coronal brain sections were mounted in stacks for the entire hypothalamus [between bregma 0.62 and −1.94 mm; Paxinos et al. (26)] on poly-l-lysin-coated glass slides. The coronal stacks were dissected into the hypothalamus and cortex, also encompassing the hippocampus (cortex/hc) for PCR and Western blot analysis (∼35 mg for the hypothalamus; ∼50 mg for the cortex/hc) and stored at −55°C for RNA extraction or protein isolation.
Western blot.
Tissue was homogenized on ice for protein isolation in a lysis buffer (50 mM Tris-HCl, 2 mM EDTA, and 1% Nonidet) with a phosphatase inhibitor (cat. P2714, P5726; Sigma-Aldrich, St. Louis, MO). Supernatants were collected and centrifuged for 15 min (6,000 g) and then stored at −55°C. A Bradford protein assay (cat. B6916; Sigma-Aldrich) was used for the determination of the protein concentration.
Samples were mixed (1:1) with 1 × Laemmli sample buffer (cat. 161-0737; Bio-Rad, Munich, Germany), containing 50 μl β-Mercaptoethanol (cat. 161-0710; Bio-Rad) per 1 ml Laemmli buffer and then incubated at 95°C for 5 min. Then, Precision Plus Protein Western C Standards (4 μl, cat. 161-0376; Bio-Rad Laboratories, Hercules, CA) and 50 μg of protein were loaded on a 10% acrylamide gel and electrophoresed at 120 V for 2 h. Thereafter, the protein was transferred (15 V, 3 mA/cm2, 60 min) to a nitrocellulose membrane (cat. 162-0150; Bio-Rad), according to the manufacturer's instructions for semidry transfer and washed with 0.1% Tween 20 in 1 × Tris-buffered saline (cat. 170–6435; Bio-Rad; TBST) three times for 5 min. Membranes were further incubated with blocking solution (10% nonfat dry milk in TBST) for 2 h at room temperature (RT), followed by application of the first antibodies overnight at 4°C [goat-anti-COX-2, 1:300 (cat. sc-1747, Santa Cruz Biotechnology, Dallas, TX); goat-antinuclear factor (NF)κB (p65), 1:400 (cat. sc-372-G, Santa Cruz Biotechnology); rabbit-anti-β-actin, 1:5,000 (cat. 2066; Sigma-Aldrich Chemie, Steinheim, Germany); and for protein ladder detection, streptavidin horseradish peroxidase (HRP; cat. 161-0380, Bio-Rad)] diluted in TBST and 5% nonfat dry milk. Thereafter, membranes were washed (3 times for 5 min) with TBST and incubated for 2 h at RT with the secondary antibodies (HRP-conjugated donkey-anti-rabbit, 1:2,000, cat. 7074, New England Biolabs, Frankfurt, Germany, or HRP-conjugated donkey-anti-goat, 1:2,000, cat. sc-2020, Santa Cruz Biotechnology), which were diluted in TBST and 5% nonfat dry milk. Subsequently, membranes were washed twice in TBST and once in TBS and incubated for 5 min with the Clarity Western ECL Substrate Kit (cat. 170–5060; Bio-Rad). Molecular Imager (Chemi-Doc XRS Imaging System; Bio-Rad) was used for 5–15 min to detect chemiluminescence. A ratio of trace quantity is presented: COX-2/β-Actin or NFκB/β-Actin multiplied by 100. Band sizes were 42 kDa for β-Actin, 72 kDa for COX-2, and 69 kDa for NFκB.
Real-time PCR.
We used Trizol-total RNA extraction (Invitrogen, Carlsbad, CA) for the hypothalamic and cortical/hippocampus (hc) tissue according to the manufacturer's protocol. One microgram of RNA was employed for reverse transcription followed by quantitative real-time PCR using TaqMan Universal PCR Master Mix with preoptimized primer/probe mixture (StepOnePlus Real-Time PCR; TaqMan Gene Expression Assay; Applied Biosystems) as previously described (7). Assay IDs for analyzed genes were as follows: GAPDH, 4352339E-1009032; COX-2, Mm00478374_m1; mPGEs, Mm00452105_m1; IκBα, Mm00477798_m1. In a previous study, GAPDH was chosen as the best housekeeping gene for normalization of cDNA quantities out of the 12 most commonly used housekeeping genes for the same type of mouse brain tissue (15) using the Primer Design Perfect Probe Norm 12 gene kit mouse and the GeNorm software (cat. ge-PP-12-mo; Primer Design, Southampton, UK). Samples were run in duplicates, and values were calculated using the comparative cycle threshold (ΔΔCT) method as x-fold difference from a control sample [sample from control group PBS, value determined as 1].
Analysis of plasma cytokines.
Serum was tested for apolipoprotein A-I (Apo A-I), CD40 (CD40), CD40 ligand (CD40-L), C-reactive protein (CRP), endothelin-1 (ET-1), eotaxin, epidermal growth factor mouse (EGF mouse), factor VII, fibrinogen, fibroblast growth factor 9 (FGF-9), fibroblast growth factor basic (FGF-basic), granulocyte chemotactic protein-2 mouse (GCP-2 mouse), growth-regulated alpha protein (KC/GRO), haptoglobin, immunoglobulin A (IgA), interferon gamma (IFN-gamma), interferon gamma-induced protein 10 (IP-10), interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta), interleukin-10 (IL-10), interleukin-11 (IL-11), interleukin-18 (IL-18), interleukin-2 (IL-2), interleukin-3 (IL-3), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-7 (IL-7), leukemia inhibitory factor (LIF), lymphotactin, macrophage colony-stimulating factor-1 (M-CSF-1), macrophage inflammatory protein-1 alpha (MIP-1 alpha), macrophage inflammatory protein-1 beta (MIP-1 beta), macrophage inflammatory protein-1 gamma (MIP-1 gamma), macrophage inflammatory protein-2 (MIP-2), macrophage inflammatory protein-3 beta (MIP-3 beta), macrophage-derived chemokine (MDC), matrix metalloproteinase-9 (MMP-9), monocyte chemotactic protein 1 (MCP-1), monocyte chemotactic protein 3 (MCP-3), monocyte chemotactic protein-5 (MCP-5), myeloperoxidase (MPO), myoglobin, oncostatin-M (OSM), stem cell factor (SCF), T-cell-specific protein, RANTES, thrombopoietin (TPO), tissue factor (TF), tissue inhibitor of metalloproteinases 1 mouse (TIMP-1 mouse), tumor necrosis factor alpha (TNF-alpha), vascular cell adhesion molecule-1 (VCAM-1), vascular endothelial growth factor A (VEGF-A), and von Willebrand factor (vWF) by a multiplexed fluorescent bead-based immunoassay (Luminex, Myriad RBM, Austin, TX). Briefly, this method operates with capture antibodies, which are attached to fluorescent microspheres and then measured by flow cytometry. All assays have been validated in a multiplex, based on immunoassay principles and tested for the parameters of the least detectable dose, precision, cross-reactivity, linearity, spiked recovery, correlation to an alternative method, dynamic range, and matrix interferences.
Statistical analysis.
All data are expressed as means ± SE. Differences between preexercise and postexercise were compared with a repeated measures ANOVA. If significant main effects were observed, post hoc analysis was conducted by using the Newman-Keuls multiple comparison test. Pearson's correlation analysis was used to identify any significant relationships between MHT and COX-2 expression or COX-2 expression and IL-6. In all cases, P < 0.05 was accepted as being significant. Data were analyzed using Prism 5 and SPSS 22 statistical analysis program.
Body weight and exercise capacity.
In all groups, body weight increased significantly from the pretraining to the posttraining time point without any significant differences between the groups (Table 1). Regarding exercise capacity, the V̇o2 max of mice from the endurance group increased significantly after training (P < 0.05, Fig. 2A), while no significant changes were observed in mice from the SG or the CG. Similarly, MHT increased significantly only in mice from the ST group after training (P < 0.05), whereas no significant changes were observed in the other groups (Fig. 2B).
Expression of inflammatory mediators in the brain.
COX-2 mRNA expression in the cortex/hc of mice from the EG increased significantly after training compared with CG (P < 0.01). There was no significant difference in the COX-2 mRNA expression in the hypothalamus after training (Fig. 3, A and B). A significant increase in COX-2 protein levels was observed in both the cortex/hc and the hypothalamus of mice from the SG (P < 0.05/P < 0.01, Fig. 3, C and D) compared with CG. NFκB protein levels were significantly increased in mice of both exercise groups in the hypothalamus (P < 0.05), while no changes were found in the cortex/hc (Fig. 3, E and F) compared with CG.
Expression of mPGES mRNA, the downstream enzyme of COX-2 required for PGE2 synthesis, was not significantly altered in the cortex/hc, while a significant increase in expression was found in the hypothalamus after both endurance as well as strength training (P < 0.01/P < 0.05, Fig. 4, A and B) compared with CG. In contrast, inhibitor of (I) κBα-mRNA expression, a negative regulator of NFκB signaling and a marker of NFκB activation (18, 38), was only significantly increased in the cortex/hc in mice of the EG (P < 0.05), but not in the hypothalamus (Fig. 4, C and D) compared with CG.
Expression of cerebral cytokines, markers of mitochondrial stress and signaling pathways.
To analyze the role of potential intracerebral signaling pathways or mediators of COX-2 activation, expression of several cytokines, markers of mitochondrial stress and signaling pathways, were analyzed in the cortex/hc and the hypothalamus. No altered expression levels were found for the cytokines interleukin (IL)-1β, IL-6, IL1 receptor antagonist (ra), the mitochondrial stress markers mitochondrial transcription factor A (TFAM), nuclear respiratory factor 1 (NRF1), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α), and the signaling molecules suppressor of cytokine signaling (SOSC3) and NF-IL6 after both training regimes (data not shown).
Levels of systemic cytokines.
To analyze a potential role for peripherally released cytokines in COX-2 expression in the brain, a panel of 50 proinflammatory, antiinflammatory, hematopoietic, chemotactic, and coagulative plasma cytokines were analyzed. While many cytokines were not affected in their basal expression levels after training, most inflammatory factors, such as IL-1α, IL-18, MIP1γ, MIP-2, IL-2, IP-10, RANTES, FGF-9, MMP9, and TIMP, decreased after training. Other proteins, like IL-7 and myoglobin, decreased in the EG, while they were upregulated in the SG after training. In both exercise groups an upregulation of IL-6 was found in mice from the EG and the SG (Fig. 5).
Association between COX-2 expression, exercise capacity, and cytokines.
A positive association was found between MHT and COX-2 expression in the cortex/hc in mice from the ST group (r = 0.39, P < 0.05). Furthermore, an association was found between COX-2 mRNA expression in the cortex and plasma IL-6 levels after endurance exercise (r = 0.37, P < 0.05).
We have shown that long-term strength and endurance training increases COX-2 expression in the cortex and hippocampus of mice. In the hypothalamus, strength training affected the COX-2 pathway including inflammatory signaling via NFκB. Both training regimes exhibited mainly antiinflammatory responses in the circulation, with an increase in IL-6 levels after training, suggesting a role for circulating IL-6 in COX-2 induction in the brain.
COX-2 expression in the brain has mostly been investigated in pathological contexts such as fever or inflammatory diseases (36, 38). In this regard, it was shown that COX-2 expression is activated by IL-6 followed by the production of PG and the generation of fever. IL-6 acts to induce PG via an IL-6 receptor mediated activation of the Janus kinase signal transducer and activator of transcription 3 (JAK-STAT3) pathway in brain endothelial cells (9, 35, 37, 44). Recent evidence suggests that acute bouts of exercise also affect the cyclooxygenase pathways in the brain (19). Others have shown that an acute bout of treadmill exercise temporarily increases COX-2 expression in the hippocampus of rats (14). The increase in COX-2 in the hippocampus was positively correlated to the results of an inhibitory avoidance test suggesting a relationship between COX-2 expression and aversive memory after exercise (19). Several other studies supported a general role of COX-2 expression in synaptic plasticity and spatial learning (19, 32, 40, 44) in addition to its function in mediating brain inflammation (18). It was also shown that regular exercise training, in addition to acute exercise, improves cognitive function by increasing brain plasticity because of an increase in neurogenesis and synaptogenesis (10, 42). This knowledge supports the results of the current study demonstrating that COX-2 expression was increased after a period of training. In particular, strength training did increase COX-2 in the cortex/hc and the hypothalamus. The pronounced effects of strength training on COX signaling are supported by the results of Portugal et al. (31), who demonstrated that not only endurance but also strength or resistance training mediates increases in cognitive function.
Interestingly, we found a positive correlation between training and COX-2 expression. Therefore we speculate that the increased COX-2 expression is part of a general adaptive response to the training. While it is known that strength training primarily results in muscular adaptation processes, it might also affect brain signaling. Mechanistically, we assume that a more efficient exercise stimulus induces a stronger increase of growth factors or other potential mediators of adaptation (25). However, since we did not measure improvement of learning ability or brain plasticity, we cannot exclude that the observed changes in brain signaling would impair learning and memory processes. In this regard, it was shown that restraint caused stress-induced alterations of the oxidative status in rat brains, which is accompanied by COX-2 induction and potential damaging results (20), although we did not detect any change in markers of mitochondrial stress in the present study (PGC1α, NRF1, TFAM). Data suggest that the ambivalent role of COX-2 on brain plasticity and learning might be related to the magnitude of COX-2 induction. While a strong stress stimulus might result in a negative physiological response, a controlled moderate stimulus given by exercise might induce a positive physiological response (28). However, since we did not find any significant regulation of NFκB or mPGES in the cortex/hc, we suggest that both long-term exercise regimes represent moderate stress stimuli, which are distinct from the acute exercise effects. In general, exercise is suggested to be a quantifiable physiological stressor. Thereby, each single bout of acute exercise might induce a moderate inflammatory process that has long-lasting effects on brain signaling (43). Similarly, adaptation processes in other tissues, like muscles, are referred to inflammatory signaling pathways (1, 12).
In the hypothalamus, levels of inflammatory signals seem to be higher in response to strength training since, here, upregulations of COX-2, NFκB, and mPGES were found. The hypothalamus represents an autoregulatory region of the brain, which is involved in the cardiorespiratory response to exercise (21). Circulating mediators like cytokines can gain access to the hypothalamus via a leaky blood brain barrier in the so-called circumventricular organs (CVOs). Some of the CVOs are located in the hypothalamus surrounding the third ventricle (34, 38). During acute exercise, the autonomic nervous system reacts and participates in the maintenance of homeostasis. After regular exercise training, the sympathetic nervous system adapts and decreases its activity (23). However, whether these functional changes are mirrored by the signaling changes observed in our study remains speculative. The inflammatory changes in the hypothalamus may simply be a result of repeated challenges of homeostasis and circulating mediators.
Changes in several plasma cytokines and growth factors were analyzed to determine the systemic inflammatory response and potential mediators of COX-2 regulation following exercise training. Interestingly, levels of most inflammatory cytokines were not affected by strength or endurance training. In line with previous studies, both exercise modes induced mainly antiinflammatory effects indicated by a decrease of proinflammatory cytokines like IL-1α, IL-18, or chemokines like RANTES (22, 30, 33).
Some other cytokines, like IL-7 and myoglobin, are differently regulated between the strength and endurance exercise groups. Endurance training significantly downregulated these proteins, while strength training significantly upregulated their expression. It is suggested that the increase of myoglobin is a result of exercise-induced muscle damage known to occur following strength training or eccentric exercise training (25, 27). Since muscle damage was not accompanied by a systemic inflammatory process, it is unlikely that it affects brain signaling. The upregulation of IL-7 after strength training is suggested to be part of the release of growth factors, which is regularly reported after strength or resistance training (12). Our finding that both types of exercise upregulated IL-6 and COX-2 and a positive association was found between systemic IL-6 and COX-2 expression indicates that circulating IL-6 is possibly involved in the regulation of COX-2 in the brain following exercise training. An increased expression of IL-6 in response to acute and chronic exposure to exercise has been shown by others (30). It is suggested that the contracting skeletal muscle releases IL-6 into the circulation (27, 29). On the basis of previous data (16) it is assumed that both training regimes might affect body composition by increasing the relative proportion of muscle tissue. Therefore the IL-6 release might be amplified in the training groups. In contrast to the release of IL-6 during inflammatory conditions, it is suggested that the IL-6 released during exercise exhibits primarily antiinflammatory effects (29). This finding might explain why most systemic inflammatory factors were downregulated after exercise training.
The connection between systemic IL-6 and brain-COX-2 expression was investigated during experimental conditions of fever (9, 37, 44). Here, it was shown that circulating IL-6 acts on endothelial cells in the brain followed by an increased mPGES and COX-2 induction. mPGES-1 is known to be an enzyme for PGE2 induction downstream of COX-2 (36). Therefore IL-6 might also be a potential mediator of COX-2 and mPGES expression after exercise training. However, since our findings are mainly descriptive and we did not find any significant change in SOCS3-mRNA expression, a potential activation marker of the IL-6-stimulated JAK-STAT3-signaling pathway (18, 38), these connections remain speculative.
Taken together, our findings demonstrate that strength training and endurance training induce COX-2 expression in the brain. The induction of COX-2 expression in the cortex/hc and hypothalamus appears to be more pronounced following strength training. The induction of these signals seems to be independent from systemic inflammatory processes because most inflammatory cytokines were not upregulated after training. A potential mediator of COX-2 expression after training might be the systemically released IL-6. However, further research is necessary to elucidate the role of inflammatory pathways on brain plasticity after endurance and strength exercise training.
No conflicts of interest, financial or otherwise, are declared by the author(s).
K.K., F.C.M., and C.R. conception and design of research; K.K. and J.B. performed experiments; K.K., J.B., and C.R. analyzed data; K.K., F.C.M., and C.R. interpreted results of experiments; K.K. and C.R. prepared figures; K.K. drafted manuscript; K.K., F.C.M., and C.R. edited and revised manuscript; K.K., F.C.M., and C.R. approved final version of manuscript.
We would like to thank D. Marks and D. Ott for excellent technical assistance and Dr. L. Harden for editing the manuscript.
- Copyright © 2016 the American Physiological Society


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20 Of The Best Signs From Planned Parenthood Rallies Happening Across the Country
Anti-abortion protests demanding the government defund of one of the nation's leading providers of affordable health care sparked nationwide counter protests Saturday as reproductive rights advocates rallied in cities across the country in defense of Planned Parenthood. Energized by the election of Donald Trump, a man who has vowed to overturn Roe v. Wade, anti-abortion activists have launched an aggressive effort to push for more restrictive abortion legislation at both national and state levels. In turn, however, they've also energized abortion rights advocates intent on defending Planned Parenthood.
In an effort to urge legislatures to defund Planned Parenthood, a national anti-abortion coalition known as #ProtestPP claimed to have organized 225 protests – spread over 45 states – against the health care provider this weekend. But Planned Parenthood has a lot of friends. And many of those friends showed up to counter protest #ProtestPP.
"[#ProtestPP rallies] are designed to intimidate patients seeking basic health care services at Planned Parenthood," Mary Alice Carter, interim vice president of Planned Parenthood Federation of America, said in a statement released Saturday. "Patients don't go to Planned Parenthood health centers to make a political statement. They go to receive quality care from a provider they know and trust. That's what we're focused on today and every day."
In celebration of Planned Parenthood, here are 20 of the best signs spotted at Defend Planned Parenthood rallies around the country:
1. If My Uterus Shot Bullets
Pro-life and pro-choice demonstrators rally outside #PlannedParenthood Peoria Health Center on Saturday.https://t.co/3jcEP1osfC pic.twitter.com/vHNNWa10Gw— Ron Johnson (@RojoPix) February 11, 2017
One sign spotted at a Planned Parenthood support rally in Peoria, Illinois read, "If my uterus shot bullets you wouldn't bother to regulate it."
2. Bitch, It's My Uterus
Protest game is on point, Silicon Valley! #indivisible #StandWithPP #PlannedParenthood @IndivisibleTeam #ProChoice pic.twitter.com/e1npvlBV7h— IndivisibleOrchard (@OCindivisible) February 11, 2017
"Bitch, it's my uterus," read one sign decorated with hearts and flowers at an I Stand With Planned Parenthood rally in California.
3. Uteruses Before Duderuses
#PlannedParenthood Rally, San Antonio, TX pic.twitter.com/qWOwMa4W8S— Ariana Garcia 🇵🇷 (@Ariana_noGrande) February 11, 2017
The sign game was strong at a Planned Parenthood support rally in San Antonio, Texas, where signs read "uteruses before duderuses" and "vaginas are more regulated than guns."
4. I Am Not Ovary-Acting
#PlannedParenthood protest has turned into a support rally after opposition group left. pic.twitter.com/iooKwVtcid— Cresencio Rodriguez (@cres_guez) February 11, 2017
A group of Planned Parenthood advocates – including one woman carrying a sign that read "I am not ovary-acting" – reportedly outnumbered those who had showed up for an anti-Planned Parenthood demonstration in Fresno, California.
5. A Free Woman
More arriving for the @PPFA rally #PlannedParenthood #MoonlightBeach #Encinitas pic.twitter.com/0QstbtUHjn— Alan Bleiweiss (@AlanBleiweiss) February 11, 2017
"No woman can call herself free who doesn't control her own body," read one Planned Parenthood supporter's sign at a rally in Encinitas, California.
6. Fallopians 4:28
#IStandWithPP today in Ferndale, Mi. The few that came to #ProtestPP found themselves mostly alone. #PlannedParenthood = #WomensHealth pic.twitter.com/BamvV5efyr— Michal Nodel (@m_adam) February 11, 2017
At an I Stand With Planned Parenthood rally in Ferndale, Michigan, a woman held a sign bearing a tongue-in-cheek spin on biblical verses – Fallopians 4:28 "Thous shalt not mess with women's reproductive rights."
7. No Uterus? No Opinion
Favorite sign at Roseville CA Planned Parenthood rally & 10 times the size of the anti group @CaliDemos @TheDemCoalition #PlannedParenthood pic.twitter.com/z3s9kiFCov— JanB (@janb2006_jan) February 11, 2017
A woman at a Planned Parenthood support rally in Roseville, California made her disdain for male legislatures' attempts to regulate women's bodies known.
8. God Bless Planned Parenthood
#plannedparenthood #persist #Neverthelessshepersisted #resist @elizabethforma @plannedparenthood pic.twitter.com/qX6JLAIvKd— Andrea Poole (@APoole313) February 11, 2017
What's better than one clever I Stand With Planned Parenthood rally signs? Three clever signs touting the greatness of Planned Parenthood!
9. Think Outside My Box
#plannedparenthood #msu pic.twitter.com/BYgyijCdsD— loretta (@LorettaCham) February 11, 2017
"Think outside my box," one woman at a Planned Parenthood rally in Michigan asked legislatures.
10. Love. Resist. Persist.
Today I defended #PlannedParenthood, because I believe in women's access to all healthcare, including abortion. #Resist #NotMyPresident pic.twitter.com/W88xShCggo— The Researcher (@JoanTheProfound) February 11, 2017
One woman used a beautiful illustration of Lady Liberty and Lady Justice to show her support for Planned Parenthood's life-saving work.
11. Get The Elephant Out Of The Womb
Set 3 of photos from #defendPP rally in Washington Sq Park. Videos at https://t.co/tolXRfTh3X #PlannedParenthood pic.twitter.com/Ob3UEW7dh1— Tardigrade CERN (@StopMotionsolo) February 11, 2017
A supporter carried a sign emblazoned with "get the elephant out of the womb" at a Defend Planned Parenthood rally held in New York City's Washington Square Park.
12. A Woman's Place Is In Your Face
Cause "a woman's place is in your face" #PlannedParenthood #nyc pic.twitter.com/iT1jvBNNLF— Leen Karadsheh (@Karadsheh3) February 11, 2017
Another sign spotted at the Defend Planned Parenthood rally in New York City's Washington Square Park reads "a woman's place is in your face."
13. Alternative Facts
Grand Rapids MI #PlannedParenthood rally. Arrow sign rules! #ActualFacts not #AlternativeFacts@PPofMI @PPact @PPFA pic.twitter.com/zuwzbxnnIK— David Orlowski (@davidorlo) February 11, 2017
In Grand Rapids, Michigan, one Planned Parenthood supporter made good use of an arrow and the Trump administration's "alternative facts" argument at a demonstration countering an anti-abortion protest.
14. Separate Cooch And State
#IStandWithPP #PlannedParenthood pic.twitter.com/FdL7ID309h— Amy Curry (@acurrydish) February 11, 2017
One Planned Parenthood advocate demanded separation between cooch and state at a rally in Fort Collins, Colorado.
15. I See What You Did There
Pun with @PPact. #StandWithPP #IStandWithPP #PlannedParenthood #healthcareforeveryone #womenshealth #womensrights #punsagainsttrump pic.twitter.com/CeKtjwSt5l— Rick Navarro (@RickNerdvarro) February 11, 2017
One man opted to fight for Planned Parenthood with puns, urging people to "get in-vulva-ed" and "be estrogen-erous with donations" to the health care provider.
16. Electile Dysfunction
Nope, not sorry for giving a f*ck. #StandWithPP #PlannedParenthood #electiledysfunction #leggomyeggos pic.twitter.com/hwqEiKjLmZ— Jaime Bauer (@j__bau) February 11, 2017
One woman gave a name to male legislatures' need to regulate women's bodies at an I Stand With Planned Parenthood rally – "electile dysfunction."
17. I Love My IUD
We who #StandWithPP far outnumbered those tryna defund PP today in Poughkeepsie #ilovemyiud #mybodymychoice #DefendPlannedParenthood @PPact pic.twitter.com/g7WCALLj7u— UCanCallMeAlexandria (@RealAlexWojcik) February 11, 2017
Just as thousands of kids once loved their MTV, thousands of women love their IUDs, as one women at a Defend Planned Parenthood rally in Poughkeepsie, New York, noted.
18. Hex Sexists
Witchy women @PPact @ppmn pic.twitter.com/1M1JCi9lc8— Deer (@Winning_Deer) February 11, 2017
A few politically conscious witches toting signs that read "hex sexists" and "witches for reproductive freedom" came out to stand with Planned Parenthood in Minneapolis, Minnesota.
19. Keep Your Theology Off My Biology
Some good signs at the #PlannedParenthood rally on Valencia pic.twitter.com/mBKCGxYi8X— Megan O-R (@meg_orp) February 11, 2017
One woman demands legislatures keep religion out of health care at a rally to support Planned Parenthood in San Francisco, California.
20. That Awkward Moment
Good morning Minnesota @ppmn @PPact pic.twitter.com/BIQ6ymolNB— Deer (@Winning_Deer) February 11, 2017
One Planned Parenthood supporter at a rally in Minneapolis, Minnesota, got right to the point with a sign that read, "That awkward moment when you care more about a potential person than an actual person."

subset: commoncrawl


text:
Next meeting
The next meeting will be held on 22 September at Melbourne Law School.
For more information please contact Network Project Officer Julian Laurens.
The next meeting will be held on 22 September at Melbourne Law School.
For more information please contact Network Project Officer Julian Laurens.
Details of past meetings and related documents will be placed here.
The Network held a meeting at Flinders University Law School, Adelaide, South Australia. The meeting was hosted by LEAD Co-convenor Tania Leiman, Associate Dean at Flinders Law School.
The Network held a meeting at the University of Notre Dame Australia (Fremantle) hosted by Lead Co-convenor Joan Squelch, Associate Dean at Notre Dame Law.
The Network held a meeting at Melbourne Law School hosted by Judith Marychurch . The meeting was well attended with representatives from 20 Australian law schools. Topics discussed included the LACC review of the Priestley 11 for the purposes of admission to the legal profession; developing the professional relationship between LEAD, CALD and other Networks/Organisations focused on legal education and research; assessment practices to maintain academic integrity; policy and curriculum responses that can meet challenges posed by the changing job market; challenges posed by online learning.
The Network held a meeting at UNSW Law School convened by Alex Steel. A key focus was on issues of statutory interpretation.
The Network held a meeting at UNSW Law School convened by Alex Steel. The meeting generated considerable ideas for Network activities in 2015, including developing surveys and studies of benefit to legal education.
The Network held a meeting at Bond University Law School on the Gold Coast Convened by Melissa de Zwart. Particular topics explored included teaching statutory interpretation, the role of technology in the classroom, technology as an aid in grading, and assessment, grade distribution and feedback. A presentation on the Smart Casual project was given by Assoc Prof Mary Heath.
A Network meeting was held at the Queensland University of Technology Law School in Brisbane. Convened by Judith McNamarra, discussion included Honours, Masters and the AQF and the internationalisation of the law school curriculum.
A Network meeting was held at the ANU College of Law in Canberra to coincide with the annual ALTA conference. Convened by Alex Steel, key discussion areas included the role of LSSSE surveys in Australia, assessment and grade distribution and Masters degrees and the AQF.
The Law Curriculum Futures Roundtable is hosted by the Law AD Netowrk and UNSW Law. It will be held from 10am to 4pm at UNSW Law School. The Guest Speaker is Prof Judith Wegner, Burton Craige Professor of Law, University of North Carolina, Distinguished Legal Education Fellow UNSW Law. The Topic was 'Curriculum review/reform across Australia and in comparison to the US'.
This networking event took place in the National Wellness for Law Forum to canvass the extent to which law schools nationally are engaging with student wellness issues. The forum also launched the CALD guidelines by Prof Paula Baron. Details of the forum are here: http://wellnessforlaw.com/.
This was a networking event within the Teaching Research Nexus in Law: Opportunities and Challenges. Members are invited to join the symposium in particular: Collaborative Workshop: Participate in a brainstorm of legal education issues in need of further exploration, and a discussion of participants' areas of research interest, with the intention of facilitating further collaborative projects. The aim of this was for the network to capture issues considered relevant to our colleagues nationally. This assisted the network to support research into legal education.
A Law AD Network Meeting was held at the Monash University Law Chambers (Melbourne CBD) on Thursday 6 December 2012 from 9am-5pm. The theme was: 'Mapping and Designing Contemporary Law Curriculum: Sharing Practice'. View agenda.
In July 2012 the Network meeting was arranged to coincide with the ALTA conference at the University of Sydney. The focus of the meeting was assessing legal skills and discused the development of methods for skills assessment in law school.
In November 2011 Bond University hosted the AD Network meeting which focussed on student wellbeing issues in the law curriculum.


text:
What are five things I should know about The Ritz-Carlton Shanghai, Pudong?
Answers from Our Experts (1)
Before you book your stay at The Ritz-Carlton Shanghai, Pudong, here are five things you should know about the Forbes Travel Guide Five-Star hotel:
1. Despite being popular with business travelers, the luxury hotel is family-friendly, offering a kids menu at Italian restaurant Scena and a big pool that's just under five feet deep. But the best parental perk by far is the free babysitting services.
2. The hotel is located half a block from a line 2 Metro station. It's one of the most well-connected lines on Shanghai's easy-to-navigate subway system.
3. The Ritz's bar Flair, located on the 58th floor, offers a large terrace, signature cocktails and sweeping views of the Pudong skyline and the Bund. The hot spot stays jam-packed all week long.
4. The hotel's lounge on the Club Level floor is open 24 hours a day and offers free-flowing drinks and food service fives times a day.
5. The gym at the Shanghai hotel is also open 24 hours a day and is often empty. So you can work out whenever you please, with plenty of room to spread out.


text:
Mark Steve
E-tech software's
Sub: Application for the post of IT Executive
Respected Sir,
This is with reference to the job opening in the IT sector of your company published on the internet on one of the placement portal. I, Joseph Millan, am interested in the job position of IT Executive and would like to present my resume in the context.
I am a recent IT graduate who demonstrates an effective combination of engineering, communication and documentation expertise, which are equally required in your job posting. I have completed my graduation in the field of Software application. I have also undertaken training in the same to sharpen my skills.
Please find the resume enclosed with the cover letter to have a look at my achievements during my training and college studies that helps you to have a clear opinion about my performance and capability.
During my studies, I have been selected as best student for many leadership duties and also during the training period awarded the best trainer award for my exceptional working skills and learning technology.
I am a go-getter and believe to achieve the targets in the given time-period. I have good communication and inter-personal skills
I will wait for a positive reply from your side.
Joseph Millan


text:
Fun Bullet Point Time, Hooray!
- Mike Hart stated a few fun things in a chat with ESPN.com people in EPSN's retarded SportsNation things. Most interestingly, he stated that He (capitalized because He is a god) and Hene were coming back "because we've gotta beat O-State." In the same interview-like ubstance, he said that the hardest hitter in the country is UM's pwn Dave Harris (ed note: he is right).
- In the same interview-like substance-like material, Leon Hall said that the wolverine D was going to hit JDB hard in the Rose Bowl because "somebody's got to pay for what happened." In a related note, John David Booty officially hates Urban Meyer and Gary Danielson.
- Re: BCS mess (and yes, I can reply to my own post), all this confusion about the BCS, voting, etc. just goes to show that perhaps the best (or even only viable?) type of poll for college football would be one like the BlogPoll. Voters are held accountable for the way they vote, there is an intelligent proprietor, and GASP! people actually watch the games! Thinking about the virtues of BlogPoll really makes me want to participate next year.
- Coming soon to a blog near you!: Rose Bowl previews, mega-early 2007 season previews, and much, much more! I'll probably get back into this "blogging" thing a bit more, now that school crunch time and Varsity Blue (the show, not the blog, silly) are over.
Posted under Analysis
Tags: Uncategorized
Comments are closed.
More Blog Post
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text:
|Reviews for Poison
| Guest chapter 13 . 9/15/2016
I found this story recently i am glad i found it. It gave a clear idea about what if itachi did was done by his own intension, the reasons have been very much satisfied. The death of itachi felt more reasonable like this than the real manga but still u made ap second scenario stating tht if he is alive was something i like to ship. Thinking of the years tht ran after u updated, feels like u wont continue it anymore. Still i felt it is important to remind u about how unique ur piece of work, its beautiful. I hope u continue the second scenario in the future.
| Hinatta chapter 1 . 3/15/2015
Another long Hinata story!
I'll add this story to my Hinata Community
PM me to be staff or if you know of any others!
Look for it here: ...
| Hinata's shadow chapter 13 . 1/14/2014
It was really good T.T
You are gonna make it to a itahina story aren't you.
Please come back quickly and do the next chapter
| Midnight Rising chapter 13 . 10/16/2012
Please continue! I can tell things are heating up!
| kandita chapter 13 . 6/4/2012
I am begging you to update! This story is just too too wonderful to just collect dust and go unfinished are even updated...
The entire story was just amazing:)! You did a great job! And I truely with all my heart hope u do update this
There is so much questions I have reading this ESPECALLLY the ending
I hope that Hinata finds out that Itachi is alive:)! I truely hope so
Ugh! I can't take this you really need to update...
Also nice pace!
| yuki25 chapter 13 . 2/22/2011
so when is the next update? I love the story wish that u can still continue with the story. I want to see how strong or any changes in hinta now that she has sharingan.. Tnx
| hell's angel chapter 13 . 1/3/2011
a hero's gonna save me just in time
this story is kind of weird but i find it very cool.i mean hinata having the uchiha bloodline is where the story starts to get intresting,but i like the cold hearted neji better and not the mushy neji. please update soon.
| aoilove chapter 13 . 12/5/2010
whau!i dont usally read itahinas (i am a sasuhina fan)but this one is fantastic...i love the way you made it get out of the rel story witout making it look stupid or too excentric...i wanna kow how it continous!pleaseplease ONEGAII!hahahaha no...you can do that ever you want but it woul be amaizing if you could contine it
| Somber Lambency chapter 13 . 7/12/2010
Great story, great author. :)
| lovingo0Kawaii0oGirl chapter 13 . 5/20/2010
please continue!
| nwyd chapter 13 . 1/18/2010
ohmygosh i love your story! i was nearly crying at the end. please continue!
| DAngel7 chapter 1 . 11/25/2009
Interesting ;] I like it!
| Leilani-chan chapter 13 . 11/16/2008
wowzers, you put tobi in it! i love tobi!
| HinaMisa chapter 1 . 5/1/2008
0-0 i SUPPORT HINA x ITA
| kunaigalHINATA chapter 8 . 4/7/2008
is that the real story of Uchiha?


text:
Just wanted to let you know I am working on the elements of Chi Running and I am already noticing a difference.
Beginner runner stays pain-free with ChiRunning.
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Website Design: JuneAllison
Website Development: PaleoSun


text:
ACCELL is focused on research and research training. We are proud of our Doctor of Philosophy (PhD), Doctor of Education (EdD), and Master degree research degree students who come from various disciplines (e.g., psychology, education, human resources, IT, engineering) and professional backgrounds (e.g., teacher, psychologist, manager, counsellor, engineer, trainer).
Current Confirmed Doctoral Candidates
- Cristy Bartlett
- David Bruce
- Pamela Hawkswell
- Jennifer Luke
- Nicole McDonald
- Brad McLennan
- Kate McMillan
- Mark Oliver
- Greg Souvan
Current Provisional EdD and PhD Doctoral Candidates and Master Degree Researchers
- Rebecca Black
- India Bryce
- Jason Brown
- Melanie Evangelista
- Naomi Griffin
- Michael Healy
- Clayton Lawrence
- Krisi Lovric
- Tessa McCredie
- Emily McDonald
Doctoral Graduates
- Matthew Capp, EdD
- Susan Carter, PhD
- Allison Creed, PhD
- Jonathan Green, PhD
- Nadine Hamilton, EdD
- Szuzsanna Horvath, PhD
- Jameel Hasaan, EdD
- Janet Buchan, PhD


text:
First the sad news.... Mason's team, the Blue Dawgs, lost their game in the semi finals today. The other team had two boys that didn't miss a shot the whole game! The final score was 35 to 42. The Blue Dawgs end the season in a respectable position of 3rd place.
Exciting news..... Clark tried out and made the Highschool Jazz Band! He is the only fish trombonist that made it into one of the 5 trombone chairs. He is also the only trombonist to make it from second band. The other four trombonist are all in first band. He is super excited!
In other news...... Here are two photos that the band photographer posted on her website. They were taken at the Alamodome on November 29, 2008. Original Post


text:
hi leed...
firstly ....are you wanting to use just one pc on the broadband ???
if so then you will first of all, have to set up wired, so that you can do all the configurations to the router...not really a lot to do as its a sky router (username and password are already set on a sky router) ...once you have a successful connection with an ethernet wire, disconnect ethernet from pc or router and plug in the usb adapter allow windows to set it up and you should then get a connection ..you then need to login to connect to the router (if you have setup the wpa protection ) using the key on the side of the router ...or on the card that came with the router.
if you need to make any changes in the router always revert to wired to do them ...as you will find wireless will lose the connection and you will not be able to complete the changes .
hope this helps ...Dave.


text:
Saints boost key defensive/draft stocks
St Kilda will head to November's AFL Draft with two picks inside the top 10 and four inside the first 45.
4:04pm Oct 19, 2017
Draft position strengthened
St Kilda has enhanced it's already strong draft position, securing Port Adelaide's second round selection in this year's AFL Draft.
2:16pm Oct 19, 2017
Austin becomes a Saint
St Kilda Football Club has secured promising key defender Logan Austin.
11:25am Oct 19, 2017


text:
Brought in to transform the playing style at Adelaide, the Spaniard is confident of achieving his mission, but has warned fans the transformation will not happen overnight
New Adelaide United coach Josep Gombau wants to bring attractive football to Hindmarsh Stadium, but has asked for patience as he seeks to implement change.
Spaniard Gombau has arrived in the A-League from Hong Kong club Kitchee, having previously worked as a youth coach in the famed Barcelona academy system.
Much is expected of a man who earned his coaching badges through the Catalonian Football Federation, and the 37-year-old wants to deliver, but said he needs time to get the job done.
"We want to play beautiful football and hope that people enjoy [it] when they come to see us," Gombau said.
"I need to start something from the beginning, which is the style, the skills. If you want to play this way you need to go step by step.
"Of course I want to win, to try and be successful, like every coach at every single club - but step by step. If you make a good step at the beginning you put yourself in a better position."
Having touched down in Australia, Gombau oversaw his first training session on Monday, and is looking forward to working with the playing group during the months ahead.
"I saw things I like a lot, and I also saw things I need to rearrange, but this is normal when arriving at a new team," he said.
"Tomorrow we will have a meeting with the players and explain what we want to do during the pre-season.
"We have a long pre-season, this is good, we have two-and-a-half months and we can go step by step and I am happy from my first day of working here."
Gombau also talked up the Spanish signings captured by Adelaide ahead of the 2013-14 campaign.
"One [Sergio Cirio] is a striker who can play in the three positions at the top," Gombau said.
"This season he played more of a left winger than other positions, and is a player that has scored a lot of goals [24] this season. I think the A-League is a league that is physically very strong, and he is a strong player.
"The other [Isaias Sanchez] player is a midfield - defensive - that is good at recovering the ball, but is also very good with the first pass. He is a player that can start an attacking play, and can play at [number] 6, 8 or 10 [on the field]."


text:
Re: Fixed star alignments (October 23, 2005)--addendum
Expand Messages
- Mark that's two references to sex in one post, "the tip of Orions
sword" and the "crotch" reference... I guess I'll have to keep my eyes
open on the news this week... or maybe not, maybe its in reference
to "at last" and maybe that means good news for me...
- I do have a burnable rom at home, just no software.
That jpsplit looks interesting, I'll have to look into that.
And actually, I just got moved into a higher department where I do the same
things but spend on average 10 minutes out of the hour working. So as
soonas I finish copying out the Keys (lol) I am going to start working on
Christian Astrology since those two are probably the hardest/longest reads
of all the books I have.
>From: "Martina" <merskine2@...>
>Reply-To: email@example.com
>To: <email@example.com>
>Subject: Re: [thefixedstars] Re: Complex astrology
>Date: Tue, 8 Nov 2005 11:56:49 -0500
>You know- CD burners are pretty cheap these days,... (as low as 24.00)
>even the CD-DVD burners are cheap,.. (under 50.00 ,..) might be worth
>thinking of upgrading,... I lost my hardrive to a virus, and of
>course hadn't backed up for a long time, because XP is so stable,..
>Just think what you'll lose in personal work if your hardrive dies
>either physically or because its infected? They are easy to install
>BTW,... I've replaced a few of mine and installed them over the years.
>Oh,.. and hard drives just die all by themselves, ya know,.. had that
>to the last one,...
>as for net access,.. you can always try
>one of the free trials at AOL,.. I noticed in the bank yesterday- they had
>the AOL disks for a free 1000 hours sitting there for the taking.
>If only to download some goodies! If you have access to the net at work
>you can split a zip file to fit on floppies,.. its called HJSplit and its
Your message has been successfully submitted and would be delivered to recipients shortly.


text:
Illumination at the Morton Arboretum
ComEd is a presenting sponsor of Illumination, an outdoor event on a one-mile, gently rolling paved path with interactive, 100% LED lighting displays. See dazzling projections, trees that respond to touch and sound, and vivid colors throughout. To redeem the buy one, get one ticket free offer, purchase online at the Morton Arboretum website with "COMED2016" discount code or print a virtual coupon and redeem at the Visitor Center Desk at the Arboretum. * Offer valid November 18-January 2 * Closed Thanksgiving Day, Christmas Eve, Christmas Day, and the following Mondays: November 21, November 28, December 5 and December 12 * Hours of Operation: Doors open at 5p.m., Last entry at 8:30p.m., Lights out at 9:30p.m. * For more information, visit the Morton Arboretum website Learn More For information on taking advantage of these offers, visit ComEd.com/HolidayLights
Event Cost: Ticket prices vary for days and peak hours. Member adult: $13-$17 Member child: (2-17) $7-$9 Nonmember adult $18-$22 Nonmember child (2-17 years old) $12-$14
Event times: Hours of Operation: Doors open at 5p.m., Last entry at 8:30p.m., Lights out at 9:30p.m.


text: I love my dog to death. She is my happiness in this world. A day ago, I created an Instagram account under her name. I will be posting videos and pictures of her every day. She already has 61 followers! She might be scared of strangers and often barks and growls at them, but I know she was meant to be famous! LOL. I always see other Shih Tzu's on Instagram and think, my doggo is way cuter than they are. Why don't I start a new Instagram for her? So I did. I take tons of pictures and videos of her so posting new things daily won't be a problem. She's so loving and loves belly rubs! She loves cucumbers and cooked vegetables and chicken. She's a traveller! I'm guessing she hates planes??? Also, she doesn't really like kids. On second thoughts, she doesn't really like anyone at all other than my immediate family and fiancé. She barks at everyone else lol. Look at that face!!!!!!


text:
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text:
The Painted Brick Building Sides of buildings in Wyoming's towns and cities, and sometimes from other areas of the West. An examination of old style advertising. . . as it looks today.
Friday, September 29, 2017
Railroad side buildings, Greeley Colorado
Posted by Pat, Marcus & Alexis at 9:51 PM
No comments:
Labels: Colorado, Greeley Colorado
Location: Greeley, CO, USA
Tuesday, September 12, 2017
Lex Anteinternet: Some things don't bear the test of time
Lex Anteinternet: Some things don't bear the test of time: The former Budget Inn in Rawlins, Wyoming. It was probably pretty modern looking when built, I'd guess in the 1950s or early 1960s. ...
Sunday, September 10, 2017
Bicycle Path, Casper Wyoming
Posted by Pat, Marcus & Alexis at 5:12 PM
No comments:
Labels: Casper, Street Markings, Wyoming
Location: E 15th St, Casper, WY, USA
Tuesday, August 22, 2017
Rotary Sidewalk Clock, Casper Wyoming
This is a newly installed Rotary Club sidewalk clock in Casper, Wyoming. This was photographed during Casper's Eclipse Festival which drew a large crowed. The clock is located at the town's new downtown plaza.
Clocks like this are sponsored by Rotary Clubs all over the country, but this is the first such clock in Casper Wyoming, and only the second sidewalk clock in Caspser of which I'm aware. I'd never seen an actual example prior to this and was surprised by how large they are. Very nice attractive clock.
Posted by Pat, Marcus & Alexis at 8:06 AM
No comments:
Labels: 2010s, Casper, Clocks, Other sidewalk features, Wyoming
Wednesday, August 9, 2017
Occident Flour, Fairview Montana
This building in Fairview, Montana advertises "Occident Flour" which apparently maintained it "costs more--worth it". I don't know what the building itself was used for. This is located in the wheat growing region of Montana.
Posted by Pat, Marcus & Alexis at 9:31 PM
No comments:
Labels: AGH Photographs, Fairview Montana, Montana
Location: Fairview, MT 59221, USA
Tuesday, July 25, 2017
Urban Bottle, (Pacific Fruit Warehouse), Casper Wyoming
This is the old Pacific Fruit Warehouse which as been remodeled as Rocca's Sicilian restaurant and Urban Bottle, a liquor store. Urban Bottle features a new painted sign resembling those that were common in southern Europe, with that look no doubt being intentional.
Posted by Pat, Marcus & Alexis at 9:35 PM
No comments:
Labels: 2010s, Liquor Store, Produce Warehouse, restaurants, Warehouse
Toyota Body Shop, Casper Wyoming
This photo is proof that not all "ghost signs" are from before World War Two. The old Toyota body shop in downtown Casper Wyoming, which had this sign painted on it during the 1970s.
Posted by Pat, Marcus & Alexis at 9:31 PM
No comments:
Utility Box, Casper Wyoming
This utility box has been on this site before from when it featured downtown historic Casper buildings. Now it features native fish.
Posted by Pat, Marcus & Alexis at 9:28 PM
No comments:
Labels: Casper, Utilities Box, Wyoming
Saturday, June 10, 2017
Morey Mercantile Company Building, Denver Colorado
This building, and the one to the immediately left, were owned by the C. S. Morey Mercantile Company. I'm not sure of its vintage or what its used for today.
Posted by Pat, Marcus & Alexis at 8:23 PM
No comments:
Labels: Colorado, Denver Colorado, retail stores.
Inkmonstr mural, Denver Colorado.
I have no idea what the story behind this is, but this Inkmonstr mural is located in downtown Denver, Colorado.
Posted by Pat, Marcus & Alexis at 8:12 PM
No comments:
Labels: Denver Colorado, Mural
Monday, June 5, 2017
Spratlen-Anderson Wholesale Grocery Company (Edbrooke Lofts), Denver Colorado
This is the Spratlen-Anderson Wholesale Grocery Company Building, and neighboring buildings, in downtown Denver Colorado. Today this it the Edbrooke Lofts. The building was constructed in 1905 with a floor added in 1911. The grocery company was dissolved in 1923 after which the Davis Brothers Drug Company occupied the company until 1957. It was converted to lofts in 1990.
Posted by Pat, Marcus & Alexis at 9:54 PM
No comments:
Labels: 1900s, 1910s, Colorado, Denver Colorado, grocery stores
Location: 1450 Wynkoop St, Denver, CO 80202, USA
The M. J. O'Fallon Plumbing Company (Colorado Saddlery) Building, Denver Colorado
This building in Lower Downtown Denver was built for the M. J. O'Fallon Plumbing Supply Company whose painted bricks still adorn it. The most famous industry to occupy this 1906 structure, however, was Colorado Saddlery which only moved out of it in 2005 after having occupied it since 1945. Today it houses shops and lofts.
Posted by Pat, Marcus & Alexis at 9:44 PM
No comments:
Labels: 1900s, Colorado, Denver Colorado, Manufacturing, Warehouse
"LoDo Wellness Center", Denver Colorado
I've been going by this giant mural of a guitar playing cowboy for years and I have in mind that it used to be a Sheplers store in downtown Denver. Now, of course, in weedy Denver it's apparently a marijuana dispensary. Oh well.
Posted by Pat, Marcus & Alexis at 9:31 PM
No comments:
Labels: Colorado, Denver Colorado, Mural, retail stores.
Tuesday, May 23, 2017
Wax Trax, Denver Colorado.
This is a photograph of Wax Trax, the legendary Denver record story located in two side by side old buildings in that city. Currently its adorned with a large painting of Lemmy Kilmister, the late front man for the band Motorhead.
This photo was obviously taken from a distance, and inside a vehicle, so the large mural isn't easily visible in the photo but it was taken from a well known photograph of Kilmister in which he's wearing a short brimmed hat featuring a U.S. Army crossed saber device and an Ordinance branch device. Kilmister had a thing for military paraphernalia.
Posted by Pat, Marcus & Alexis at 6:30 AM
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Labels: Denver Colorado, retail stores.
Sunday, May 14, 2017
Painted Bricks: Purina Elevator, Denver Colorado
A new elevator mural:
Purina Elevator, Denver Colorado:Hmmm. . . puppy replaced by a kitten. . . symbolic?
Terrible photo, but the elevator features a puppy playing with a ball.
Posted by Pat, Marcus & Alexis at 7:48 AM
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Labels: Colorado, Denver Colorado, elevators
Sunday, April 23, 2017
Sonic Rainbow, Casper Wyoming: Independent Record Store Day, 2017
This store has featured here before. This is the Sonic Rainbow record store in Casper Wyoming. Every year it features new original window art for Independent Record Store Day. On this occasion, its patrons were lined up early in the morning waiting for it to open for that event.
The crowd would grow larger on this rainy morning. These people were there early.
Posted by Pat, Marcus & Alexis at 5:52 AM
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Labels: Casper, retail stores., Windows, Wyoming
Thursday, March 16, 2017
Lex Anteinternet: 1917 The Year that made Casper what it is. Or ma...
Lex Anteinternet: 1917 The Year that made Casper what it is. Or ma...: I have no before and after pictures for Casper that would cleanly show what the town looked like in January, 1916 and then later looked lik...
Posted by Pat, Marcus & Alexis at 6:55 AM
No comments:
Labels: 1910s, Blog Mirror, World War One
Monday, January 30, 2017
Alley mural, Casper Wyoming.
Posted by Pat, Marcus & Alexis at 8:56 PM
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Event listener performance with vanilla JS
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text:
|But some premies were so nice ....
|Re: Re: I think that we weren't selfish enough to be true premies -- Thorin
|Post Reply
|Top of thread
Posted by: Jean-Michel ®
02/28/2003, 13:27:41
I vividly remember an episode back in the late 70s in the ashram. I was that ashram secretary/housefather at that time, and one premie (woman) started to have serious mental problems. (That was in Bry sur Marne, for the French readers). I won't give her name. She's since deceased from cancer.
I couldn't understand at that time how an ashram premie could have mental problems. That was beyond my understanding, I had a feeling that something was wrong in paradise - maybe that was one of my first drips. But I was a full time employee of DLM at that time, and I felt it was my responsibility to take care of her. Nobody else would do anything for her, even discuss the issue ! I used to take her back and forth to the hospital where we was getting treatment. I don't remember all the details, but I was the only one taking care of her.
She finally left the ashram and moved back to her family. Someone managed to convince her that the ashram was not the right place for her.
Modified by Jean-Michel at Fri, Feb 28, 2003, 13:29:14
|Post Reply | Recommend | Alert
|Where am I? Original Top of thread
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text:
Home :Products :Cat Machines, Work Tools and Attachments :Non-current Cat Machines :Excavators :321C LCR
The C Series incorporates innovations for improved performance and versatility.
Engine Model Cat® 3066T ATAAC
Net Flywheel Power 137 hp
Net Power - ISO 9249 138 hp
Bore 4.02 in
Stroke 5.12 in
Displacement 389 in3
Net Power - EEC 80/1269 138 hp
Net Power - SAE J1349 137 hp
Operating Weight 51864 lb
Operating Weight - Minimum 50416 lb
Operating Weight - Std. Undercarriage 51863.69 lb
Counterweight 13448.18 lb
Maximum Drawbar Pull 44063 lb
Maximum Travel Speed 3.4 mph
Hydraulic System
Main Implement System - Maximum Flow (2x) 54 gal/min
Max. Pressure - Equipment 4974 psi
Max. Pressure - Travel 4974 psi
Max. Pressure - Swing 3625 psi
Pilot System - Maximum Flow 11 gal/min
Pilot System - Maximum Pressure 566 psi
Boom Cylinder - Bore 4.92 in
Boom Cylinder - Stroke 55 in
B1 Family Bucket Cylinder - Bore 4.72 in
B1 Family Bucket Cylinder - Stroke 41 in
Service Refill Capacities
Fuel Tank Capacity 87 gal
Cooling System 8.5 gal
Engine Oil 7.9 gal
Swing Drive 2.1 gal
Final Drive (each) 2.6 gal
Hydraulic System (including tank) 69 gal
Swing Mechanism
Swing Speed 10.6 RPM
Swing Torque 45582 lb ft
Standard w/Long Undercarriage 32 in
Optional 24 in
Optional 28 in
Ground Clearance - Fixed Gauge 18.7 in
Transport width 125.2 in
Compact Radius
Compact radius design delivers top performance in tight quarters.
Compact Radius Design
The 321C LCR features a compact radius, making it ideal for working in tight areas such as: next to buildings, highway construction - limiting lane closures, logging roads and other tight areas where space is restricted.
Operator Confidence
Due to the 321C LCR's compact working envelope, operators can work in confidence that the counterweight will not swing into any object behind them, which allows the operator to concentrate on the task at hand.
Working Envelope
The 321C LCR has been designed to rotate with little to none of the counterweight extending beyond its tracks, and working within its width. To further minimize the working envelope, the 321C LCR's boom is positioned further back in the upper frame as compared to a standard excavator. This reduces the front swing radius when the boom is pulled all the way up and the stick is brought back.
Engine and Hydraulics
The Cat 3066T engine and hydraulics give the 321C LCR exceptional power, efficiency, and controllability unmatched in the industry for consistently high performance in all applications.
Six cylinder turbocharged engine built for power, reliability, economy and low emissions will keep the machine up and running. The Cat 3066T engine meets Tier 2 worldwide emissions requirements.
Automatic Engine Speed Control
The two-stage one touch control maximizes fuel efficiency and reduces sound levels.
Low Sound, Low Vibration
The Cat 3066T design improves operator comfort by reducing sound and vibration.
Electronic Control Module
The Electronic Control Module (ECM) works as the "brain" of the engine's control system, responding quickly to operating variables to maximize engine efficiency. Fully integrated with sensors in the engine's fuel, air, coolant, and exhaust systems, the ECM stores and relays information on conditions such as rpm, fuel consumption, and diagnostic information.
Hydraulic Cross Sensing System
Improves productivity with faster implement speeds and quicker, stronger pivot turns.
Fine Swing Control
Fine swing control cushions swing start and stop for better implement control.
Hydraulic Cylinder Snubbers
The hydraulic cylinder snubbers at rod-end of boom cylinders and both ends of stick cylinder cushion shocks, reduce sound, and increase cylinder life, keeping the machine working longer.
The hydraulic system offers precise control to the 321C LCR, reducing operator fatigue, improving operator effectiveness and efficiency, which ultimately translates into enhanced performance.
Boom and Stick Regeneration Circuit
Boom and stick regeneration circuit increases efficiency and reduces cycle times for higher productivity and lower operating costs.
321C LCR structural components and undercarriage are the backbone of the machine's durability.
Robotic Welding
Up to 95% of the structural welds on a Caterpillar hydraulic excavator are completed by robots. Robotic welds achieve up to three times the penetration of manual welds.
Carbody Design and Track Roller Frames
X-shaped, box-section carbody provides excellent resistance to torsional bending. Robot-welded track roller frames are press-formed, pentagonal units that deliver exceptional strength and service life.
Main Frame
Rugged main frame is designed for maximum durability and efficient use of materials.
Durable Cat undercarriage absorbs stresses and provides excellent stability.
Rollers and Idlers
Sealed and lubricated track rollers, carrier rollers, and idlers provide excellent service life, that keeps the machine in the field longer.
Long Undercarriage
The long (L) undercarriage maximizes stability and lift capacity. This long, wide, and sturdy undercarriage offers a very stable work platform.
Booms and Sticks
Built for performance and long service life, Caterpillar booms and sticks are large, welded, box-section structures with thick, multi-plate fabrications in high stress areas.
Reach Boom
The reach boom features an optimum design that maximizes digging envelopes.
R2.9B Stick
The R2.9B stick is versatile and used in a variety of applications.
Operator Station
The 321C LCR operator workstation is quiet with ergonomic control placement and convenient adjustments, low lever and pedal effort, ergonomic seat design and highly efficient ventilation.
Cab Design
An enlarged cab with curved styling gives the operator a comfortable, spacious working environment and excellent visibility.
Sliding Door
The cab door slides alongside the cab and takes less space to open and close than a hinged door. This unique design allows the operator to easily get in and out of the cab when working against walls on job sites, even when attachments are added.
New, compact monitor enhances viewing while displaying a variety of easy to read and understand language-based information.
Climate Control
The 321C LCR features a semi-automatic climate control with air intake system. The air conditioner is standard and adjusts temperature and flow.
The upper front windshield opens, closes, and stores below the roof of the operator with a one-touch action release system. Grips on the mid-lower portion of the front windshield assist in easy opening.
The enlarged right-side window provides excellent visibility and isolates the operator from the hydraulic lines. The upper cab door window slides open, providing ventilation and allowing the operator to easily communicate with people on the work site.
The pop-up skylight provides improved upward visibility and opens easily with the assistance of a gas cylinder.
Work Tools - Attachments
The 321C LCR provides greater versatility by offering factory installed auxiliary hydraulics, couplers and a variety of bucket types and sizes.
Quick Couplers
The Pin Grabber Plus and the Dedicated Hydraulic Quick Coupler enhances machine versatility by enabling the rapid change over of a wide range of work tools in the field.
General Purpose (GP) Buckets
General Purpose Buckets are best for digging in soft to hard ground with low to moderately abrasive materials.
Ditch Cleaning (DC) Buckets
These wide shallow buckets are best for bank forming, ditch cleaning, and finishing.
Heavy-Duty Rock (HDR) Buckets
Heavy-Duty Rock Buckets perform best when digging fragmented rock, frozen ground, caliche, and highly abrasive materials.
Heavy-Duty (HD) Buckets
Heavy-duty buckets are for digging in moderate to hard material and feature large ground engaging tools, thick cutting edges and thick bottom and side wear plates to improve performance in demanding conditions.
Heavy-Duty Power (HDP) Buckets
Designed to improve breakout force and machine cycle times, the Heavy Duty Power Bucket (HDP) compliments the General Purpose, Heavy Duty, and Heavy Duty Rock bucket lines.
With the optional Tool Control System, up to five different tool settings may be pre-programmed and selected from the electronic controller through the monitor.
Work Tools
Choose from a variety of work tools such as hammers, shears, thumbs, rotators, grapples, or crushers. Ask your Cat dealer for information on attachments or special configurations.
Tool Control System
The optional Tool Control System maximizes work tool productivity by configuring hydraulic flow, pressure and operator controls to match a specific work tool. System versatility enables a wide range of tools to be used. Factory installed hammer and thumb circuits are also available as attachments.
Simplified service and maintenance save you time and money.
Extended Service Intervals
Extended service and maintenance intervals reduce service time and machine availability. Use of oil-free bearing extends front linkage greasing interval to 1000 hours, except in the bucket area, which has been extended to 100 hours.
Ground-Level Maintenance
For operator convenience, daily maintenance areas can be easily reached from ground level.
Pre-Start Monitoring System
This system allows the operator to check coolant, hydraulic oil and engine oil levels from the monitor inside the cab.
Anti-Skid "Pressed-Star" Plate
The top of the storage box and surface of the upper structure are covered with "pressed-star" plate to prevent the service person or operator from slipping during maintenance.
Handrail and Steps
Larger handrails and steps make it easier for the operator to climb on and off the machine.
Fan Guard
Engine radiator fan and oil cooler are completely enclosed by fine wire mesh, reducing the risk of injury.
Greasing Points
A concentrated remote greasing block on the boom delivers grease to hard-to-reach locations.
Service Doors
Service doors are located on both sides of the upper structure. The doors make it easy to reach maintenance items such as the engine radiator or any hydraulic components from ground level. Openings are large and service doors latch in the full open position.
Air Filter
The air filter features a double-element construction for superior cleaning efficiency. When the air cleaner plugs, a warning is displayed on the monitor screen inside the cab.
Fuel-Water Separator
The water separator has a primary fuel filter element and is located in the radiator compartment for easy access from the ground.
Complete Customer Support
Cat dealer services help you operate longer with lower costs.
Make detailed comparisons of the machines you are considering before you buy. What are the job requirements, machine attachments and operating hours? What production is needed? What is the true cost of lost production? Your Cat dealer can give precise answers to these questions.
For the best operating techniques to increase productivity and your profit, turn to your Cat dealer for the latest training literature and trained staff.
Repair option programs guarantee the cost of repairs up front. Diagnostic programs such as Scheduled Oil Sampling and Technical Analysis help you avoid unscheduled repairs.
Repair, rebuild or replace? Your Cat dealer can help you evaluate the cost involved so you can make the right choice.
Product Support
You will find nearly all parts at our dealer parts counter. Cat dealers utilize a worldwide computer network to find in-stock parts to minimize machine downtime. You can save money with Cat remanufactured components.
Your local Cat dealer is there to support and protect you. Extended warranty options are also available.
Standard Equipment
- Alternator, 50 Ampere
- Working Light
- Horn
Operator Environment
- AM/FM Radio with Speakers
- Ashtray
- Beverage Holder
- Air Conditioner, Semi-automatic
- Coat Hook
- Door, Sliding
- Floor Mat, Washable
- Hydraulic Neutralizer Lever for All Controls
- Joystick Type Controls, Pilot Operated
- Language Display Monitor with Gauges
- Laminate Front Windshield
- Lighting, Interior
- Literature Holder
- Pop-up Skylight, Polycarbonate with Sun Shade
- Retractable Front Windshield with Assist Device
- Seat, Suspension Type
- Storage Compartment
- Tempered Windows
- Travel Control Pedals with Footrests
- Windshield Wiper with Washer, Upper
Power Train
- Cat 3066T Diesel Engine
- 2300 m (7500 ft) Altitude Capability without Deration
- Air-to-Air Aftercooling (ATAAC)
- Automatic Engine Speed Control
- Cooling
- Straight Line Travel
- Two Speed Auto-shift Travel
- Water Separator in Fuel Line
- Hydraulic Track Adjusters
- Idler and Center Section Track Guiding Guards
- Track-type Undercarriage with Grease Lubricated Seals
Other Standard Equipment
- Automatic Swing Brake
- Automatic Work Modes
- Auxiliary Hydraulic Valve (1)
- Capability of Stackable Valve for Main Valve
- Capability of Auxiliary Circuit
- Counterweight 6100 kg (13,448 lb)
- Door Locks and Cap Locks with One-key Security System
- Pre-wired for Product Link Capability
- System Mirrors
- Wave Fin Radiator
Optional Equipment
- Alarm, Travel
- Light, Working, Cab Mounted
- Light, Boom, Right Side
- Power Supply, 12V-5A (1 socket)
Operator Environment
- Cab with Polycarbonate Windows
- Hand Control Pattern Changer
- Seat, Suspension Type
Power Train
- Cold Weather Starting Aid
- Guard, Track Guiding
- Guard, Track Guiding
- Track Shoes
Other Optional Equipment
- 3 Auxiliary Hydraulic Arrangement Options
- Hammer Circuit
- Thumb Circuit
- Tool Control Circuit
- Boom, Reach 5.68 m (18 ft 7 in)
- Control, Fine Swing
- Coupler, Hydraulic Pin Grabber
- Linkage, Bucket, B-Family
- Drive, for Auxiliary Pump
- Guard, Swivel
- Guard, Vandalism
- Guard, Full Front Windshield
- Stick, 2.9 m (9 ft 6 in)
Larry Cummins
Ed Rottmann
865-595-1060 or 865-599-8225
Larry Cummins
Ed Rottmann
865-595-1060 or 865-599-8225
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Cat 246
my cat 246 was running fine when we parked it . the next day turned the key on, had buzzer, glow plu... [More]
my cat 246 was running fine when we parked it . the next day turned the key on, had buzzer, glow plu... [More]
CAT 307C 2004 Hydraulic Drawings
I've got an older colleague with a 307C 2004-model excavator. He came to me asking if it's possible... [More]
I've got an older colleague with a 307C 2004-model excavator. He came to me asking if it's possible... [More]
Over Excitation Shutdown Problem For 3512 Diesel Generator
please give any advice to me.. i have using caterpillar 3512 generator from last 4 months in my... [More]
please give any advice to me.. i have using caterpillar 3512 generator from last 4 months in my... [More]
Fuel Lines
I need help I have a D4D dozer 1974 year model and the s/n 82j3434 . I bought it ran great went back... [More]
I need help I have a D4D dozer 1974 year model and the s/n 82j3434 . I bought it ran great went back... [More]