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Late-life enalapril administration induces nitric oxide-dependent and independent metabolic adaptations in the rat skeletal muscle
Marzetti, Emanuele
Calvani, Riccardo
DuPree, Jameson
Lees, Hazel A.
Giovannini, Silvia
Seo, Dong-oh
Buford, Thomas W.
Sweet, Kindal
Morgan, Drake
Strehler, Kevin Y. E.
Diz, Debra
Borst, Stephen E.
Moningka, Natasha
Krotova, Karina
Carter, Christy S.
Recently, we showed that administration of the angiotensin-converting enzyme inhibitor enalapril to aged rats attenuated muscle strength decline and mitigated apoptosis in the gastrocnemius muscle. The aim of the present study was to investigate possible mechanisms underlying the muscle-protective effects of enalapril. We also sought to discern the effects of enalapril mediated by nitric oxide (NO) from those independent of this signaling molecule. Eighty-seven male Fischer 344 × Brown Norway rats were randomly assigned to receive enalapril (n = 23), the NO synthase (NOS) inhibitor NG-nitro-l-arginine methyl ester (l-NAME; n = 22), enalapril + l-NAME (n = 19), or placebo (n = 23) from 24 to 27 months of age. Experiments were performed on the tibialis anterior muscle. Total NOS activity and the expression of neuronal, endothelial, and inducible NOS isoforms (nNOS, eNOS, and iNOS) were determined to investigate the effects of enalapril on NO signaling. Transcript levels of tumor necrosis factor-alpha (TNF-α) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) were assessed to explore actions of enalapril on inflammation and mitochondrial biogenesis, respectively. Protein expression of energy-sensing and insulin signaling mediators, including protein kinase B (Akt-1), phosphorylated Akt-1 (pAkt-1), mammalian target of rapamycin (mTOR), AMP-activated protein kinase subunit alpha (AMPKα), phosphorylated AMPKα (pAMPKα), and the glucose transporter GLUT-4, was also determined. Finally, the generation of hydrogen peroxide (H2O2) was quantified in subsarcolemmal (SSM) and intermyofibrillar (IFM) mitochondria. Enalapril increased total NOS activity, which was prevented by l-NAME co-administration. eNOS protein content was enhanced by enalapril, but not by enalapril + l-NAME. Gene expression of iNOS was down-regulated by enalapril either alone or in combination with l-NAME. In contrast, protein levels of nNOS were unaltered by treatments. The mRNA abundance of TNF-α was reduced by enalapril relative to placebo, with no differences among any other group. PCG-1α gene expression was unaffected by enalapril and lowered by enalapril + l-NAME. No differences in protein expression of Akt-1, pAkt-1, AMPKα, pAMPKα, or GLUT-4 were detected among groups. However, mTOR protein levels were increased by enalapril compared with placebo. Finally, all treatment groups displayed reduced SSM, but not IFM H2O2 production relative to placebo. Our data indicate that enalapril induces a number of metabolic adaptations in aged skeletal muscle. These effects result from the concerted modulation of NO and angiotensin II signaling, rather than from a dichotomous action of enalapril on the two pathways. Muscle protection by enalapril administered late in life appears to be primarily mediated by mitigation of oxidative stress and pro-inflammatory signaling.
Aging; Nitric oxide synthase (NOS) isoforms; Mitochondria; mTOR; Glucose tolerance; l-NAME; Inflammation; ACE inhibitors
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Inhibition of NF-κB-induced inflammatory responses by angiotensin II antagonists in aged rat kidney
Kim, Ji Min
Heo, Hyoung-Sam
Choi, Yean Ja
Ye, Byeong Hyeok
Seo, Arnold Young
Yu, Byung Pal
Leeuwenburgh, Chrstiaan
Chung, Hae Young
Carter, Christy S.
Experimental gerontology
In this study, we explored the mechanisms by which the angiotensin converting enzyme inhibitor (ACEI), enalapril, and the Ang II receptor blocker (ARB), losartan suppress oxidative stress and NF-κB activation-induced inflammatory responses in aged rat kidney. The experimentations were carried out utilizing aged (24-month-old) Brown Norway x Fischer 344 (F1) male rats which were randomized into 3 groups and administered enalapril (40 mg/kg), losartan (30 mg/kg) or placebo for 6 months (daily p.o.). The level of reactive species (RS), peroxynitrite (ONOO−), GSH/GSSG and lipid peroxidation were measured. The activity of the pro-inflammatory transcription factor NF-κB, and gene expression of proteins in upstream signaling cascades were measured by electro-mobility shift assay (EMSA) and Western blotting. Enalapril and losartan differentially attenuated redox imbalance and the redox-sensitive transcription factor, NF-κB pathway. Furthermore, stimulation of the NF-κB activation pathway by phosphorylation of p65 was attenuated by both compounds. Moreover, mediation of phosphorylation of p65 by phosphorylation of IκB kinase αβ (IKKαβ) and mitogen- and stress-activated protein kinase-1 (MSK1), were also inhibited by enalapril and losartan. Finally, both compounds also lowered expression of NF-κB-dependent inflammatory genes, such as cyclooxygenase-2 (COX-2),) and inducible NO synthase (iNOS). Only losartan lowered levels of 5-lipoxygenase (5-LOX). These findings indicate that enalapril and losartan differentially suppress inflammatory responses via inhibition of oxidative stress-induced NF-κB activation in aged rat kidney.
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Differential effects of enalapril and losartan on body composition and indices of muscle quality in aged male Fischer 344 × Brown Norway rats
Carter, Christy S.
Giovaninni, Silvia
Seo, Dong-Oh
DuPree, Jameson
Morgan, Drake
Chung, Hae Young
Lees, Hazel
Daniels, Michael
Hubbard, Gene B.
Lee, Shuko
Ikeno, Yuji
Foster, Thomas C.
Buford, Thomas W.
Marzetti, Emanuele
The primary purpose of the present set of studies was to provide a direct comparison of the effects of the angiotensin-converting enzyme inhibitor enalapril and the angiotensin receptor blocker losartan on body composition, physical performance, and muscle quality when administered late in life to aged rats. Overall, enalapril treatment consistently attenuated age-related increases in adiposity relative to both placebo and losartan. The maximal effect was achieved after 3 months of treatment (between 24 and 27 months of age), at a dose of 40 mg/kg and was observed in the absence of any changes in physical activity, body temperature, or food intake. In addition, the reduction in fat mass was not due to changes in pathology given that enalapril attenuated age-related increases in tumor development relative to placebo- and losartan-treated animals. Both enalapril and losartan attenuated age-related decreases in grip strength, suggesting that changes in body composition appear dissociated from improvements in physical function and may reflect a differential impact of enalapril and losartan on muscle quality. To link changes in adiposity to improvements in skeletal muscle quality, we performed gene array analyses to generate hypotheses regarding cell signaling pathways altered with enalapril treatment. Based on these results, our primary follow-up pathway was mitochondria-mediated apoptosis of myocytes. Relative to losartan- and placebo-treated rats, only enalapril decreased DNA fragmentation and caspase-dependent apoptotic signaling. These data suggest that attenuation of the severity of skeletal muscle apoptosis promoted by enalapril may represent a distinct mechanism through which this compound improves muscle strength/quality.
Age-related adiposity; Body composition; Sarcopenia; Renin–angiotensin system; Physical function; Muscle quality
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Differential effects of late-life initiation of low-dose enalapril and losartan on diastolic function in senescent Fischer 344 × Brown Norway male rats
Groban, Leanne
Lindsey, Sarah
Wang, Hao
Lin, Marina S.
Kassik, Kimberly A.
Machado, Frederico S. M.
Carter, Christy S.
No proven pharmacological therapies to delay or reverse age-related diastolic dysfunction exist. We hypothesized that late-life low-dose (non-blood-pressure-lowering) angiotensin-converting enzyme inhibition vs. angiotensin II receptor blockade would be equally efficacious at mitigating diastolic dysfunction in the senescent Fischer 344 × Brown Norway rat. Enalapril (10 mg/kg/day; n = 9) initiated at 24 months of age and continued for 6 months, increased myocardial relaxation (e'), reduced Doppler-derived indices of filling pressure (E/e'), favorably lowered the ratio of phospholamban–SERCA2 and reduced oxidative stress markers, Rac1 and nitrotyrosine, in aged hearts. Treatment with losartan (15 mg/kg/day; n = 9) similarly mitigated signs of cardiac oxidative stress, but impairments in diastolic function persisted when compared with untreated rats (n = 7). Our findings favor the idea that the lusitropic benefit of low-dose angiotensin-converting enzyme inhibitor initiated late in life may be related to an antioxidant-mediated modulation of SERCA2, resulting in improved relaxation rather than via overt effects on cardiac structure or blood pressure.
Angiotensin-converting enzyme inhibitor; Angiotensin II receptor blocker; Diastolic dysfunction; Oxidative stress; SERCA2; Tissue Doppler
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Effect of Aging and Obesity on Insulin Responsiveness and Glut-4 Glucose Transporter Content in Skeletal Muscle of Fischer 344 × Brown Norway Rats
Larkin, Lisa M.
Reynolds, Thomas H.
Supiano, Mark A.
Kahn, Barbara B.
Halter, Jeffrey B.
The journals of gerontology. Series A, Biological sciences and medical sciences
This study investigated the metabolic changes with age in the Fischer 344 × Brown Norway rat and its suitability as an animal model of postmaturational insulin resistance. Specifically, we determined whether an age-associated decrease in glucose disposal is associated with diminished whole body insulin responsiveness and/or a decrease in glucose transporter (GLUT-4) protein and mRNA content in medial gastrocnemius muscle of male Fischer 344 × Brown Norway rats of ages 8, 18, and 28 months. Fasting plasma glucose was unchanged with age. There was a significant age effect on visceral adiposity, fasting plasma insulin levels, insulin responsiveness, and GLUT-4 protein content. Insulin responsiveness and GLUT-4 protein were lower in the 18-month-old rats than in the 8-month-old rats. The findings of age-associated increases in visceral adiposity and insulin resistance, and decreases in GLUT-4 in the Fisher 344 × Brown Norway rat, suggest that this rat strain may be an appropriate model for studying the effects of aging on glucose homeostasis.
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Heterogeneous Effects of Calorie Restriction on In Vivo Glucose Uptake and Insulin Signaling of Individual Rat Skeletal Muscles
Sharma, Naveen
Sequea, Donel A.
Castorena, Carlos M.
Arias, Edward B.
Qi, Nathan R.
Cartee, Gregory D.
PLoS ONE
Calorie restriction (CR) (consuming ∼60% of ad libitum, AL, intake) improves whole body insulin sensitivity and enhances insulin-stimulated glucose uptake by isolated skeletal muscles. However, little is known about CR-effects on in vivo glucose uptake and insulin signaling in muscle. Accordingly, 9-month-old male AL and CR (initiated when 3-months-old) Fischer 344xBrown Norway rats were studied using a euglycemic-hyperinsulinemic clamp with plasma insulin elevated to a similar level (∼140 µU/ml) in each diet group. Glucose uptake (assessed by infusion of [14C]-2-deoxyglucose, 2-DG), phosphorylation of key insulin signaling proteins (insulin receptor, Akt and Akt substrate of 160kDa, AS160), abundance of GLUT4 and hexokinase proteins, and muscle fiber type composition (myosin heavy chain, MHC, isoform percentages) were determined in four predominantly fast-twitch (epitrochlearis, gastrocnemius, tibialis anterior, plantaris) and two predominantly slow-twitch (soleus, adductor longus) muscles. CR did not result in greater GLUT4 or hexokinase abundance in any of the muscles, and there were no significant diet-related effects on percentages of MHC isoforms. Glucose infusion was greater for CR versus AL rats (P<0.05) concomitant with significantly (P<0.05) elevated 2-DG uptake in 3 of the 4 fast-twitch muscles (epitrochlearis, gastrocnemius, tibialis anterior), without a significant diet-effect on 2-DG uptake by the plantaris or either slow-twitch muscle. Each of the muscles with a CR-related increase in 2-DG uptake was also characterized by significant (P<0.05) increases in phosphorylation of both Akt and AS160. Among the 3 muscles without a CR-related increase in glucose uptake, only the soleus had significant (P<0.05) CR-related increases in Akt and AS160 phosphorylation. The current data revealed that CR leads to greater whole body glucose disposal in part attributable to elevated in vivo insulin-stimulated glucose uptake by fast-twitch muscles. The results also demonstrated that CR does not uniformly enhance either insulin signaling or insulin-stimulated glucose uptake in all muscles in vivo.
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Greater insulin sensitivity in calorie restricted rats occurs with unaltered circulating levels of several important myokines and cytokines
Sharma, Naveen
Castorena, Carlos M
Cartee, Gregory D
Nutrition & Metabolism
Calorie restriction (CR; ~60% of ad libitum, AL intake) has been associated with substantial alterations in body composition and insulin sensitivity. Recently, several proteins that are secreted by nontraditional endocrine tissues, including skeletal muscle and other tissues, have been discovered to modulate energy metabolism, body composition, and insulin sensitivity. The aim of this study was to characterize the influence of CR by rats on plasma levels of six of these newly recognized metabolic hormones (BDNF, FGF21, IL-1β, myonectin, myostatin, and irisin). Body composition of 9-month old male Fischer-344/Brown Norway rats (AL and CR groups) was determined by nuclear magnetic resonance. Blood sampled from the carotid artery of unanesthetized rats was used to measure concentrations of glucose and plasma proteins. As expected, CR versus AL rats had significantly altered body composition (reduced percent fat mass, increased percent lean mass) and significantly improved insulin sensitivity (based on the homeostasis model assessment-estimated insulin resistance index). Also consistent with previous reports, CR compared to AL rats had significantly greater plasma levels of adiponectin and corticosterone. However, there were no significant diet-related differences in plasma levels of BDNF, FGF21, IL-1β, myonectin, myostatin, or irisin. In conclusion, these results indicate that alterations in plasma concentration of these six secreted proteins are not essential for the CR-related improvement in insulin sensitivity in rats.
Myonectin; Myostatin; FGF21; Irisin; Insulin resistance; Dietary restriction; Adiponectin; FNDC5
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Iron Accumulation with Age, Oxidative Stress and Functional Decline
Xu, Jinze
Knutson, Mitchell D.
Carter, Christy S.
Leeuwenburgh, Christiaan
PLoS ONE
Identification of biological mediators in sarcopenia is pertinent to the development of targeted interventions to alleviate this condition. Iron is recognized as a potent pro-oxidant and a catalyst for the formation of reactive oxygen species in biological systems. It is well accepted that iron accumulates with senescence in several organs, but little is known about iron accumulation in muscle and how it may affect muscle function. In addition, it is unclear if interventions which reduced age-related loss of muscle quality, such as calorie restriction, impact iron accumulation. We investigated non-heme iron concentration, oxidative stress to nucleic acids in gastrocnemius muscle and key indices of sarcopenia (muscle mass and grip strength) in male Fischer 344 X Brown Norway rats fed ad libitum (AL) or a calorie restricted diet (60% of ad libitum food intake starting at 4 months of age) at 8, 18, 29 and 37 months of age. Total non-heme iron levels in the gastrocnemius muscle of AL rats increased progressively with age. Between 29 and 37 months of age, the non-heme iron concentration increased by approximately 200% in AL-fed rats. Most importantly, the levels of oxidized RNA in gastrocnemius muscle of AL rats were significantly increased as well. The striking age-associated increase in non-heme iron and oxidized RNA levels and decrease in sarcopenia indices were all attenuated in the calorie restriction (CR) rats. These findings strongly suggest that the age-related iron accumulation in muscle contributes to increased oxidative damage and sarcopenia, and that CR effectively attenuates these negative effects.
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Chronic ingestion of 2-deoxy-D-glucose induces cardiac vacuolization and increases mortality in rats
Minor, Robin K.
Smith, Daniel L.
Sossong, Alex M.
Kaushik, Susmita
Poosala, Suresh
Spangler, Edward L.
Roth, George S.
Lane, Mark
Allison, David B.
de Cabo, Rafael
Ingram, Donald K.
Mattison, Julie A.
Toxicology and applied pharmacology
Calorie restriction (CR), the purposeful reduction of energy intake with maintenance of adequate micronutrient intake, is well known to extend the lifespan of laboratory animals. Compounds like 2-deoxy-D-glucose (2DG) that can recapitulate the metabolic effects of CR are of great interest for their potential to extend lifespan. 2DG treatment has been shown to have potential therapeutic benefits for treating cancer and seizures. 2DG has also recapitulated some hallmarks of the CR phenotype including reduced body temperature and circulating insulin in short-term rodent trials, but one chronic feeding study in rats found toxic effects. The present studies were performed to further explore the long-term effects of 2DG in vivo. First we demonstrate that 2DG increases mortality of male Fischer-344 rats. Increased incidence of pheochromocytoma in the adrenal medulla was also noted in the 2DG treated rats. We reconfirm the cardiotoxicity of 2DG in a 6-week follow-up study evaluating male Brown Norway rats and a natural form of 2DG in addition to again examining effects in Fischer-344 rats and the original synthetic 2DG. High levels of both 2DG sources reduced weight gain secondary to reduced food intake in both strains. Histopathological analysis of the hearts revealed increasing vacuolarization of cardiac myocytes with dose, and tissue staining revealed the vacuoles were free of both glycogen and lipid. We did, however, observe higher expression of both cathepsin D and LC3 in the hearts of 2DG-treated rats which indicates an increase in autophagic flux. Although a remarkable CR-like phenotype can be reproduced with 2DG treatment, the ultimate toxicity of 2DG seriously challenges 2DG as a potential CR mimetic in mammals and also raises concerns about other therapeutic applications of the compound.
Deoxyglucose; Calorie restriction; Lifespan; Mortality; Cardiac vacuolarization
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The effects of aging on the functional and structural properties of the rat basilar artery
Tümer, Nihal
Toklu, Hale Z.
Muller‐Delp, Judy M.
Oktay, Şehkar
Ghosh, Payal
Strang, Kevin
Delp, Michael D.
Scarpace, Philip J.
Physiological Reports
Aging leads to progressive pathophysiological changes in blood vessels of the brain and periphery. The aim of this study was to evaluate the effects of aging on cerebral vascular function and structure. Basilar arteries were isolated from male Fischer 344 cross Brown Norway (F344xBN) rats at 3, 8, and 24 months of age. The basilar arteries were cannulated in the pressurized system (90 cm H2O). Contractile responses to KCl (30–120 mmol/L) and endothelin‐1 (10−11–10−7 mol/L) were evaluated. Responses to acetylcholine (ACh) (10−10–10−4 mol/L), diethylamine (DEA)‐NONO‐ate (10−10–10−4 mol/L), and papaverin (10−10–10−4 mol/L) were assessed to determine both endothelium‐dependent and endothelium‐independent responsiveness. Advanced aging (24 months) decreased responses of the basilar artery to both the contractile and relaxing agents; whereas, DEA‐induced dilation was significantly higher in the 8‐month‐old group compared with the younger and older groups. The arterial wall‐to‐lumen ratio was significantly increased in 24‐month‐old rats. Smooth muscle cell count was also decreased in old rats. These findings indicate that aging produces dysfunction of both the endothelium and the vascular smooth muscle in the basilar artery. Aging also alters wall structure of the basilar artery, possibly through decreases in smooth muscle cell number and concomitant hypertrophy.
The purpose of this study was to determine the effects of advancing age on the structure and vasomotor responses of the basilar artery as well as the serum antioxidant capacity. Advanced aging (24 months) decreased responses of the basilar artery to both the contractile and relaxing agents, whereas, DEA‐induced dilation was significantly higher in the 8‐month‐old group compared with the younger and older rats. The arterial wall‐to‐lumen ratio was significantly increased in 24‐month‐old rats. Smooth muscle cell count was also decreased in old rats. Our findings demonstrate that aging is associated with functional impairment in endothelium‐dependent and ‐independent relaxation responses and contractility in the basilar arteries, and these diminished responses are accompanied by structural remodeling and decreased antioxidant capacity of the serum.
Acetylcholine; aging; antioxidant capacity; basilar artery; endothelin; NO; papaverin; vascular smooth muscle; vasoreactivity; vertebrobasilar insufficiency
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Comparison of Temporal Transcriptomic Profiles from Immature Lungs of Two Rat Strains Reveals a Viral Response Signature Associated with Chronic Lung Dysfunction
Hines, Elizabeth A.
Szakaly, Renee J.
Leng, Ning
Webster, Anais T.
Verheyden, Jamie M.
Lashua, Amber J.
Kendziorski, Christina
Rosenthal, Louis A.
Gern, James E.
Sorkness, Ronald L.
Sun, Xin
Lemanske, Robert F.
PLoS ONE
Early life respiratory viral infections and atopic characteristics are significant risk factors for the development of childhood asthma. It is hypothesized that repeated respiratory viral infections might induce structural remodeling by interfering with the normal process of lung maturation; however, the specific molecular processes that underlie these pathological changes are not understood. To investigate the molecular basis for these changes, we used an established Sendai virus infection model in weanling rats to compare the post-infection transcriptomes of an atopic asthma susceptible strain, Brown Norway, and a non-atopic asthma resistant strain, Fischer 344. Specific to this weanling infection model and not described in adult infection models, Sendai virus in the susceptible, but not the resistant strain, results in morphological abnormalities in distal airways that persist into adulthood. Gene expression data from infected and control lungs across five time points indicated that specific features of the immune response following viral infection were heightened and prolonged in lungs from Brown Norway rats compared with Fischer 344 rats. These features included an increase in macrophage cell number and related gene expression, which then transitioned to an increase in mast cell number and related gene expression. In contrast, infected Fischer F344 lungs exhibited more efficient restoration of the airway epithelial morphology, with transient appearance of basal cell pods near distal airways. Together, these findings indicate that the pronounced macrophage and mast cell responses and abnormal re-epithelialization precede the structural defects that developed and persisted in Brown Norway, but not Fischer 344 lungs.
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Schaser, Allison J.
Wang, Hao
Volz, Lana M.
Connor, Nadine P.
Age-related tongue weakness may contribute to swallowing deficits in the elderly. One contributing factor may be an alteration in muscle fiber type properties with aging. However, it is not clear how muscle fiber types within the aged tongue may vary from those found in young adults, or how fiber types may vary across the anteroposterior axis of the extrinsic tongue muscles. We examined myosin heavy chain (MHC) composition of anterior, medial, and posterior sections of the genioglossus muscle (GG) in 10 old male Fischer 344/Brown Norway rats and compared findings to previously reported data from young adult male rats. Significant differences (p< .01) between young adult and old rats were found in the distribution of MHC isoforms along the anteroposterior axis of the muscle. In the anterior, medial, and posterior regions, there was a significantly smaller proportion of type IIb MHC in the old rat GG muscles, while the proportion of type IIx MHC was significantly greater. In the medial region, the proportion of type I MHC was found to be significantly greater in the old rats. Thus, we found a shift to more slowly contracting muscle fibers in the aged rat tongue.
Tongue; Dysphagia; Myosin heavy chain; Aging; Deglutition; Deglutition Disorders
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Effect of aging on rat skeletal muscle β-AR function in male Fischer 344 × Brown Norway rats
Larkin, Lisa M.
Halter, Jeffrey B.
Supiano, Mark A.
The American journal of physiology
1996;270(2 Pt 2):R462-R468.
In this study, we tested the hypothesis that, in the male Fischer 344 × Brown Norway (F344xBN) rat, aging would be associated with an increase in sympathetic nervous system activity and a decrease in skeletal muscle β-adrenergic-receptor (β-AR) density and function. Radioligand-binding studies using [125I]iodocyanopindolol were done to evaluate β-AR density (Bmax) and antagonist-binding affinity in gastrocnemius and cardiac muscle from 6-, 18-, and 28-mo-old male F344xBN rats. β-AR function was measured as adenylyl cyclase (AC) activity stimulated by the β-AR agonist isoproterenol (Iso, 10−4 M). Basal arterial plasma norepinephrine (pNE) concentrations were higher in the 28-than in the 6- and 18-mo-old rats. Bmax was greatest and Iso-stimulated AC activity was unchanged in gastrocnemius muscle of the 28-mo-old age group. In contrast, there was an age-associated decrease in Bmax and Iso-stimulated AC activity in cardiac muscle. In conclusion, there was an age-associated increase in pNE concentrations in male F344xBN rats, suggesting an increase in sympathetic nervous system activity. In addition, there was an age-associated increase in skeletal muscle β-AR density, whereas in skeletal muscle β-AR-stimulated AC activity remained unchanged with age.
adenylyl cyclase; catecholamines; sympathetic nervous system; isoproterenol; forskolin
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A persistent increase in insulin-stimulated glucose uptake by both fast-twitch and slow-twitch skeletal muscles after a single exercise session by old rats
Xiao, Yuanyuan
Sharma, Naveen
Arias, Edward B.
Castorena, Carlos M.
Cartee, Gregory D.
Exercise has been demonstrated to enhance subsequent insulin-stimulated glucose uptake (GU) by predominantly type II (fast-twitch) muscle of old rats, but previous research has not evaluated exercise effects on GU by type I (slow-twitch) muscle from old rats. Accordingly, we studied male Fischer 344/Brown Norway rats (24 months old) and determined GU (0, 100, 200, and 5,000 μU/ml insulin) of isolated soleus (predominantly type I) and epitrochlearis (predominantly type II) muscles after one exercise session. Epitrochlearis (100, 200, and 5,000 μU/ml insulin) and soleus (100 and 200 μU/ml insulin) GU were greater at 3-h postexercise vs. age-matched sedentary controls. Insulin receptor tyrosine phosphorylation (Tyr1162/1163) was unaltered by exercise in either muscle. Akt phosphorylation (pAkt) was greater for exercised vs. sedentary rats in the epitrochlearis (Ser473 and Thr308 with 100 and 200 μU/ml, respectively) and soleus (Ser473 with 200 μU/ml). AS160 phosphorylation (pAS160) was greater for exercised vs. sedentary rats in the epitrochlearis (Thr642 with 100 μU/ml), but not the soleus. Exercised vs. sedentary rats did not differ for total protein abundance of insulin receptor, Akt, AS160, or GLUT4 in either muscle. These results demonstrate that both predominantly type I and type II muscles from old rats are susceptible to exercise-induced improvement in insulin-mediated GU by mechanisms that are independent of enhanced insulin receptor tyrosine phosphorylation or altered abundance of important signaling proteins or GLUT4. Exercise-induced elevation in pAkt, and possibly pAS160, may contribute to this effect in the epitrochlearis of old rats, but other mechanisms are likely important for the soleus.
Glucose transport; Aging; Insulin signaling; Insulin resistance
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Cytokine mRNA expression profiles in rats infected with the intestinal nematode Nippostrongylus brasiliensis.
Matsuda, S
Uchikawa, R
Yamada, M
Arizono, N
Infection and Immunity
Although the immune responses to intestinal nematode infection have been well studied and have been shown to be strongly driven by Th2-associated cytokines in mice, such information has been limited with respect to rats. We investigated changes in levels of the mRNAs encoding interleukin-2 (IL-2), IL-3, IL-4, IL-5, IL-10, and gamma interferon in the mesenteric lymph nodes of rats infected with Nippostrongylus brasiliensis by reverse transcription-PCR in comparison with immunoglobulin E (IgE)/IgG2a antibody, eosinophil, basophil, and mucosal mast cell responses. In the two rat strains used, Brown Norway and Fischer-344, which show different responses to allergens, serum IgE increased to much higher levels in the former than in the latter 2 weeks after infection. Intestinal mastocytosis was observed much earlier and more intensely in Brown Norway rats than in Fischer-344 rats, but the degrees of peripheral eosinophilia and basophilia did not differ between the two strains. In both strains, IL-3, IL-4, and IL-5 mRNA expression increased and peaked around 7 to 14 days after infection, while expression of IL-2, IL-10, and gamma interferon mRNAs did not change notably throughout the experimental period. The highest IL-4 mRNA expression was observed slightly earlier in Brown Norway than in Fischer-344 rats, but levels of IL-3 and IL-5 mRNAs peaked synchronously in both strains. The amounts of mRNAs encoding these three cytokines were always higher in Brown Norway than in Fischer-344 rats. It is suggested that in rats, Th2 or Th2-like cells are also induced after nematode infection, and IgE elevation is mainly related to increased IL-4 gene expression.
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Renin-Angiotensin System Blockers Protect Pancreatic Islets against Diet-Induced Obesity and Insulin Resistance in Mice
Frantz, Eliete Dalla Corte
Crespo-Mascarenhas, Camila
Barreto-Vianna, Andre Rodrigues C.
Aguila, Marcia Barbosa
Mandarim-de-Lacerda, Carlos Alberto
PLoS ONE
The associations between obesity, hypertension and diabetes are well established, and the renin-angiotensin system (RAS) may provide a link among them. The effect of RAS inhibition on type 2 diabetes is still unclear; however, RAS seems to play an important role in the regulation of the pancreas and glucose intolerance of mice fed high-fat (HF) diet.
C57BL/6 mice fed a HF diet (8 weeks) were treated with aliskiren (50 mg/kg/day), enalapril (30 mg/kg/day) or losartan (10 mg/kg/day) for 6 weeks, and the protective effects were extensively compared among groups by morphometry, stereological tools, immunostaining, Western blotting and hormonal analysis.
All RAS inhibitors significantly attenuated the increased blood pressure in mice fed a HF diet. Treatment with enalapril, but not aliskiren or losartan, significantly attenuated body mass (BM) gain, glucose intolerance and insulin resistance, improved the alpha and beta cell mass and prevented the reduction of plasma adiponectin. Furthermore, enalapril treatment improved the protein expression of the pancreatic islet Pdx1, GLUT2, ACE2 and Mas receptors. Losartan treatment showed the greatest AT2R expression.
Our findings indicate that ACE inhibition with enalapril attenuated several of the deleterious effects of the HF diet. In summary, enalapril appears to be responsible for the normalization of islet morphology and function, of alpha and beta cell mass and of Pdx1 and GLUT2 expression. These protective effects of enalapril were attributed, primarily, to the reduction in body mass gain and food intake and the enhancement of the ACE2/Ang (1-7) /Mas receptor axis and adiponectin levels.
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Metabolomic profiling reveals severe skeletal muscle group-specific perturbations of metabolism in aged FBN rats
Garvey, Sean M.
Dugle, Janis E.
Kennedy, Adam D.
McDunn, Jonathan E.
Kline, William
Guo, Lining
Guttridge, Denis C.
Pereira, Suzette L.
Edens, Neile K.
Mammalian skeletal muscles exhibit age-related adaptive and pathological remodeling. Several muscles in particular undergo progressive atrophy and degeneration beyond median lifespan. To better understand myocellular responses to aging, we used semi-quantitative global metabolomic profiling to characterize trends in metabolic changes between 15-month-old adult and 32-month-old aged Fischer 344 × Brown Norway (FBN) male rats. The FBN rat gastrocnemius muscle exhibits age-dependent atrophy, whereas the soleus muscle, up until 32 months, exhibits markedly fewer signs of atrophy. Both gastrocnemius and soleus muscles were analyzed, as well as plasma and urine. Compared to adult gastrocnemius, aged gastrocnemius showed evidence of reduced glycolytic metabolism, including accumulation of glycolytic, glycogenolytic, and pentose phosphate pathway intermediates. Pyruvate was elevated with age, yet levels of citrate and nicotinamide adenine dinucleotide were reduced, consistent with mitochondrial abnormalities. Indicative of muscle atrophy, 3-methylhistidine and free amino acids were elevated in aged gastrocnemius. The monounsaturated fatty acids oleate, cis-vaccenate, and palmitoleate also increased in aged gastrocnemius, suggesting altered lipid metabolism. Compared to gastrocnemius, aged soleus exhibited far fewer changes in carbohydrate metabolism, but did show reductions in several glycolytic intermediates, fumarate, malate, and flavin adenine dinucleotide. Plasma biochemicals showing the largest age-related increases included glycocholate, heme, 1,5-anhydroglucitol, 1-palmitoleoyl-glycerophosphocholine, palmitoleate, and creatine. These changes suggest reduced insulin sensitivity in aged FBN rats. Altogether, these data highlight skeletal muscle group-specific perturbations of glucose and lipid metabolism consistent with mitochondrial dysfunction in aged FBN rats.
Electronic supplementary material
The online version of this article (doi:10.1007/s10522-014-9492-5) contains supplementary material, which is available to authorized users.
Muscle; Aging; Metabolomics; Sarcopenia; Biomarkers; NAD
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Changes in IL-15 expression and death-receptor apoptotic signaling in rat gastrocnemius muscle with aging and life-long calorie restriction
Marzetti, Emanuele
Carter, Christy S.
Wohlgemuth, Stephanie E.
Lees, Hazel A.
Giovannini, Silvia
Anderson, Barbara
Quinn, LeBris S.
Leeuwenburgh, Christiaan
Mechanisms of ageing and development
TNF-α-mediated apoptosis is enhanced in aged rodent muscles, suggesting that this pathway may be involved in sarcopenia. Interleukin-15 (IL-15), a muscle-derived anabolic cytokine, mitigates muscle wasting and apoptosis in cachectic rats. This effect is thought to occur through inhibition of TNF-α-triggered apoptosis. We investigated IL-15 signaling and the TNF-α-mediated pathway of apoptosis in the gastrocnemius muscle of Fischer344×Brown Norway rats across the ages of 8, 18, 29 and 37 months, in relation to life-long calorie restriction (CR, 40% calorie intake reduction). Aging caused loss of muscle mass and increased apoptotic DNA fragmentation, which were mitigated by CR. Protein levels of IL-15 and mRNA abundance of IL-15 receptor α-chain decreased in senescent ad libitum (AL) fed rats, but were maintained in CR rodents. Elevations of TNF-α, TNF-receptor 1, cleaved caspase-8 and -3 were observed at advanced age in AL rats. These changes were prevented or mitigated by CR. Our results indicate that aging is associated with decreased IL-15 signaling in rat gastrocnemius muscle, which may contribute to sarcopenia partly through enhanced TNF-α-mediated apoptosis. Preservation of IL-15 signaling by CR may therefore represent a further mechanism contributing to the anti-aging effect of this dietary intervention in skeletal muscle.
sarcopenia; interleukin-15; tumor necrosis factor-α; calorie restriction; apoptosis
Related Articles
Effect of aging on ultrasonic vocalizations and laryngeal sensorimotor neurons in rats
Basken, Jaime N.
Connor, Nadine P.
Ciucci, Michelle R.
Experimental brain research. Experimentelle Hirnforschung. Experimentation cerebrale
While decline in vocal quality is prevalent in an aging population, the underlying neurobiological mechanisms contributing to age-related dysphonia are unknown and difficult to study in humans. Development of an animal model appears critical for investigating this issue. Using an established aging rat model, we evaluated if 50-kHz ultrasonic vocalizations in 10, 32-month-old (old) Fischer 344/Brown Norway rats differed from those in 10, 9-month-old (young adult) rats. The retrograde tracer, Cholera Toxin β, was injected to the thyroarytenoid muscle to determine if motoneuron loss in the nucleus ambiguus was associated with age. Results indicated that older rats had vocalizations with diminished acoustic complexity as demonstrated by reduced bandwidth, intensity, and peak frequency, and these changes were dependent on the type of 50-kHz vocalization. Simple calls of old rats had reduced bandwidth, peak frequency, and intensity while frequency-modulated calls of old rats had reduced bandwidth and intensity. Surprisingly, one call type, step calls, had increased duration in the aged rats. These findings reflect phonatory changes observed in older humans. We also found significant motoneuron loss in the nucleus ambiguus of aged rats, which suggests that motoneuron loss may be a contributing factor to decreased complexity and quality of ultrasonic vocalizations. These findings suggest that a rat ultrasonic phonation model may be useful for studying age-related changes in vocalization observed in humans.
Aging; Voice; Nucleus ambiguus; Neurodegeneration; Ultrasonic vocalization; Rat
Related Articles
Age-related activation of mitochondrial caspase-independent apoptotic signaling in rat gastrocnemius muscle
Marzetti, Emanuele
Wohlgemuth, Stephanie Eva
Lees, Hazel Anne
Chung, Hae-Young
Giovannini, Silvia
Leeuwenburgh, Christiaan
Mechanisms of ageing and development
Mitochondria-mediated apoptosis represents a central process driving age-related muscle loss. However, the temporal relation between mitochondrial apoptotic signaling and sarcopenia as well as the regulation of release of pro-apoptotic factors from the mitochondria has not been elucidated. In this study, we investigated mitochondrial apoptotic signaling in skeletal muscle of rats across a wide age range. We also investigated whether mitochondrial-driven apoptosis was accompanied by changes in the expression of Bcl-2 proteins and components of the mitochondrial permeability transition pore (mPTP). Analyses were performed on gastrocnemius muscle of 8-, 18-, 29- and 37- month-old male Fischer344×Brown Norway rats (9 per group). Muscle weight declined progressively with advancing age, concomitant with increased apoptotic DNA fragmentation. Cytosolic and nuclear levels of apoptosis inducing factor (AIF) and endonuclease G (EndoG) increased in old and senescent animals. In contrast, cytosolic levels of cytochrome c were unchanged with age. Mitochondrial Bcl-2, Bax and Bid increased dramatically in 37-month-old rats, with no changes in the Bax/Bcl-2 ratio in any of the age groups. Finally, expression of cyclophilin D was enhanced at very old age. Our findings indicate that the mitochondrial caspase-independent apoptotic pathway may play a more prominent role in skeletal muscle loss than caspase-mediated apoptosis.
sarcopenia; apoptosis; permeability transition pore; AIF; endonuclease G
Related Articles
Increased RNA oxidative damage and iron content in skeletal muscle with aging and disuse atrophy
Hofer, Tim
Marzetti, Emanuele
Xu, Jinze
Seo, Arnold Y.
Gulec, Sukru
Knutson, Mitchell D.
Leeuwenburgh, Christiaan
Dupont-Versteegden, Esther E.
Experimental gerontology
Muscle atrophy with aging or disuse is associated with deregulated iron homeostasis and increased oxidative stress likely inflicting damage to nucleic acids. Therefore, we investigated RNA and DNA oxidation, and iron homeostasis in gastrocnemius muscles. Disuse atrophy was induced in 6- and 32-month old male Fischer 344/Brown Norway rats by 14 days of hind limb suspension (HS). We show that RNA, but not DNA, oxidative damage increased 85% with age and 36% with HS in aged muscle. Additionally, non-heme iron levels increased 233% with aging and 83% with HS at old age, while staining for free iron was strongest in the smallest fibers. Simultaneously, the mRNA abundance of transferrin receptor-1 decreased by 80% with age and 48% with HS for young animals, while that of the hepcidin regulator hemojuvelin decreased 37% with age, but increased about 44% with disuse, indicating a dysregulation of iron homeostasis favoring increased intracellular free iron in atrophied muscles. RNA and DNA concentrations increased with age and were negatively correlated with muscle mass, whereas protein concentrations decreased with aging, indicating a preferential loss of protein compared to nucleic acids. Furthermore, xanthine oxidase activity increased with age, but not with HS, while mRNA abundance of the Y box-binding protein-1, which has been suggested to bind oxidized RNA, did not change with age or HS. These results suggest that RNA oxidation, possibly mediated by increased non-heme iron, might contribute to muscle atrophy due to disuse particularly in aged muscle.
gastrocnemius; hemojuvelin (HJV); RNA oxidation; sarcopenia; transferrin receptor 1 (TfR1); Y box binding protein 1 (YB-1); xanthine oxidase (XOD)
Related Articles
Angiotensin converting enzyme inhibition ameliorates glomerular filtration of macromolecules and water and lessens glomerular injury in the rat.
Remuzzi, A
Puntorieri, S
Battaglia, C
Bertani, T
Remuzzi, G
Journal of Clinical Investigation
The effect of enalapril on glomerular hemodynamics and permselectivity and on subsequent sclerosis was studied in male MWF/Ztm rats which spontaneously develop proteinuria and glomerular structural damage. Untreated group 1 and enalapril-treated group 2 (50 mg/liter, in the drinking water) underwent micropuncture studies after 2 mo of observation. After the same period of treatment, group 3 (untreated) and group 4 (enalapril treated) were used for determination of whole-kidney function and neutral dextran clearances. Group 5 (untreated) and group 6 (enalapril treated) were followed for an additional 4 mo and used for kidney function and morphological studies. Enalapril significantly lowered systolic blood pressure, which was elevated in untreated groups, and significantly reduced proteinuria (295 +/- 64 vs. 128 +/- 24 mg/24 h by the end of the study). Despite the reduced renal perfusion pressure, whole-kidney glomerular filtration rate was higher in enalapril-treated than in untreated rats (0.96 +/- 0.14 vs. 0.81 +/- 0.10 ml/min, P less than 0.05) as was the single nephron glomerular filtration rate (54 +/- 7.1 vs. 46 +/- 4.0 nl/min, P less than 0.05). The single glomerular afferent plasma flow was comparable in both groups. Enalapril reduced mean glomerular capillary hydraulic pressure from the normal value of 51 +/- 1 mmHg (untreated rats) to a value lower than normal (44 +/- 1 mmHg, P less than 0.001). These hemodynamic changes were associated with a significant reduction in afferent (approximately 23%) and efferent (approximately 26%) arteriolar resistance. The mean ultrafiltration coefficient was two times higher in the enalapril (0.126 +/- 0.027 nl/s per mmHg) than in the untreated group (0.061 +/- 0.023 nl/s per mmHg). The clearance of dextran macromolecules relative to that of inulin was significantly reduced for all molecular sizes studied (26-64 A) in enalapril-treated vs. untreated rats. Theoretical analysis of dextran fractional clearances using a heteroporous model of neutral solute transport across the glomerular capillary wall indicated that enalapril affected glomerular membrane size selective properties, reducing uniformly the radius of hypothetical membrane pores. Enalapril treatment also significantly limited (P less than 0.01) the development of glomerular structural lesions (mean percentage of sclerotic glomeruli was 4.2 +/- 3.5% [treated] vs. 28 +/- 15% [untreated] rats at the end of the study) as well as tubulo-interstitial damage. These results suggest that the protective effect of enalapril on the development of proteinuria and glomerular sclerosis in this model is due to its property of ameliorating size selectivity and hydraulic permeability of the glomerular capillaries.
Related Articles
Impaired Bladder Function in Aging Male Rats
Zhao, Weixin
Aboushwareb, Tamer
Turner, Chanda
Mathis, Cathy
Bennett, Colleen
Sonntag, William E.
Andersson, Karl-Erik
Christ, George
The Journal of urology
The prevalence of bladder dysfunctions increases with age. In humans it is difficult to separate changes related to exogenous factors from those directly related to the aging process. Some confounding variables can be avoided by studying age related changes in an animal model. We evaluated the impact of age on bladder function in vivo and in vitro, and characterized the corresponding morphological changes.
Materials and Methods
Young (4 to 6 months old) and old (older than 28 to 30 months) male Fischer/Brown Norway rats were used in the study. Cystometric studies were done in conscious, freely moving rats. After cystometry tissue strips from the bladder body were used in in vitro studies of muscarinic receptor activation and electrical field stimulation, and histological examination.
Old rats had higher bladder weight than young rats but the bladder-to-body weight ratio did not change. We noted significant age related differences in 8 of 10 cystometric parameters. Old rats had increased bladder capacity, post-void residual volume, micturition volume and frequency, baseline and intermicturition pressure, and spontaneous activity but decreased micturition pressure. Bladder strip responses to carbachol and electrical field stimulation were significantly lower in old than in young rats. Histological examination revealed urothelial thinning, lower muscle mass and higher collagen content in the bladders of old vs young rats.
Physiological aging alters bladder function in male rats even when external factors remain constant. Thus, in old rats bladder capacity, post-void residual urine and spontaneous activity are higher, and responses to muscarinic receptor stimulation and electrical field stimulation are lower than in young rats. Such changes correspond to findings in aging human bladders, supporting the view that the Fischer/Brown Norway rat is a useful model in which to study age related bladder function changes.
urinary bladder; physiology; aging; urodynamics; male
Related Articles
Targeted Training of Ultrasonic Vocalizations in Aged and Parkinsonian Rats
Johnson, Aaron M.
Doll, Emerald J.
Grant, Laura M.
Ringel, Lauren
Shier, Jaime N.
Ciucci, Michelle R.
Journal of Visualized Experiments : JoVE
Voice deficits are a common complication of both Parkinson disease (PD) and aging; they can significantly diminish quality of life by impacting communication abilities. 1, 2 Targeted training (speech/voice therapy) can improve specific voice deficits,3, 4 although the underlying mechanisms of behavioral interventions are not well understood. Systematic investigation of voice deficits and therapy should consider many factors that are difficult to control in humans, such as age, home environment, age post-onset of disease, severity of disease, and medications. The method presented here uses an animal model of vocalization that allows for systematic study of how underlying sensorimotor mechanisms change with targeted voice training. The ultrasonic recording and analysis procedures outlined in this protocol are applicable to any investigation of rodent ultrasonic vocalizations.
The ultrasonic vocalizations of rodents are emerging as a valuable model to investigate the neural substrates of behavior.5-8 Both rodent and human vocalizations carry semiotic value and are produced by modifying an egressive airflow with a laryngeal constriction.9, 10 Thus, rodent vocalizations may be a useful model to study voice deficits in a sensorimotor context. Further, rat models allow us to study the neurobiological underpinnings of recovery from deficits with targeted training.
To model PD we use Long-Evans rats (Charles River Laboratories International, Inc.) and induce parkinsonism by a unilateral infusion of 7 μg of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle which causes moderate to severe degeneration of presynaptic striatal neurons (for details see Ciucci, 2010).11, 12 For our aging model we use the Fischer 344/Brown Norway F1 (National Institute on Aging).
Our primary method for eliciting vocalizations is to expose sexually-experienced male rats to sexually receptive female rats. When the male becomes interested in the female, the female is removed and the male continues to vocalize. By rewarding complex vocalizations with food or water, both the number of complex vocalizations and the rate of vocalizations can be increased (Figure 1).
An ultrasonic microphone mounted above the male's home cage records the vocalizations. Recording begins after the female rat is removed to isolate the male calls. Vocalizations can be viewed in real time for training or recorded and analyzed offline. By recording and acoustically analyzing vocalizations before and after vocal training, the effects of disease and restoration of normal function with training can be assessed. This model also allows us to relate the observed behavioral (vocal) improvements to changes in the brain and neuromuscular system.
Related Articles
Tissue-Specific Responses of IGF-1/Insulin and mTOR Signaling in Calorie Restricted Rats
Sharma, Naveen
Castorena, Carlos M.
Cartee, Gregory D.
PLoS ONE
Moderate calorie restriction (CR) (∼60% of ad libitum, AL, intake) has been associated with numerous favorable physiological outcomes in many species, and the insulin/IGF-1 and mTOR signaling pathways have each been proposed as potential mediators for many of CR's bioeffects. However, few studies have assessed the widely held idea that CR induces the down-regulation of the insulin/IGF-1 and/or mTOR pathways in multiple tissues. Accordingly, we analyzed the phosphorylation status of 11 key signaling proteins from the insulin/IGF-1 (IRTyr1162/1163, IGF-1RTyr1135/1136, IRS-1Ser312, PTENSer380, AktSer473, GSK3αSer21, GSK3βSer9) and mTOR (TSC2Ser939, mTORSer2448, P70S6KThr412, RPS6Ser235/236) pathways in 11 diverse tissues [liver, kidney, lung, aorta, two brain regions (cortex and cerebellum), and two slow-twitch and three fast-twitch skeletal muscles] from 9-month-old male AL and CR Fischer 344 x Brown Norway rats. The rats were studied under two conditions: with endogenous insulin levels (i.e., AL>CR) and with insulin infused during a hyperinsulinemic-euglycemic clamp so that plasma insulin concentrations were matched between the two diet groups. The most striking and consistent effect of CR was greater pAkt in 3 of the 5 skeletal muscles of CR vs. AL rats. There were no significant CR effects on the mTOR signaling pathway and no evidence that CR caused a general attenuation of mTOR signaling across the tissues studied. Rather than supporting the premise of a global downregulation of insulin/IGF-1 and/or mTOR signaling in many tissues, the current results revealed clear tissue-specific CR effects for the insulin signaling pathway without CR effects on the mTOR signaling pathway.
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text:
- Report: #950348
Report - Rebuttal - Arbitrate
Complaint Review: Bottless
BottlessPO Box 1797 Bend, Oregon United States of America
Bottless Mark Anthony Bottless and its Owner Mark Anthony required payment in full but have not delivered for 3 months! Bend, Oregon
*Author of original report: Update
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Now I cannot even get a return phone call, despite having paid in full 3 months ago! Mark Anthony and this company are either con artists or are so disorganized they can't deliver on the products for which they accept payment. I can't tell which it is, but I have suffered a total loss with no delivery for what I paid. Do not pay in advance for anything from this company or Mark Anthony!
This report was posted on Ripoff Report on 10/03/2012 01:33 PM and is a permanent record located here: http://www.ripoffreport.com/r/Bottless/Bend-Oregon-97709/Bottless-Mark-Anthony-Bottless-and-its-Owner-Mark-Anthony-required-payment-in-full-but-hav-950348. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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subset: commoncrawl


text:
1600PX: No matter how many times I have spotted this livery, I have been in absolute awe!
This elegant -800 series 747 is seen taxying out for a departure in the very first light of the day. Western Europe was experiencing very high levels of air pollution at the time and already the h... (more)
Rolling for take off, runway 18.


text:
Hi all !!
I would like to change the display of the blogroll but i don't find the file.
In my source code, i find this:

Blogoliste

I was first thinking that it was the bookmark-template.php but in the code source:
echo '
cat_ID . '" class="linkcat">
' . $cat->cat_name . "
\n\t
- \n";
so a h2 and not a h3 ...
Where can i change it please ??
Thx a lot


text:
Order Reprints
Journal: Molecules, 2010
Volume: 15
Number: 3857
Article: A New Cembrane Diterpene from the Bornean Soft Coral Nephthea sp.
Authors: Takahiro Ishii, Zhan Zhaoqi and Charles Santhanaraju Vairappan
Link: http://www.mdpi.com/1420-3049/15/6/3857
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text:
Home > Eyeglasses > Chloé > CE2659 (626)
CE 2659 is available in 3 additional attractive styles
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Mehr zur CE2659/626 Eyewear
The new Chloé underlines that you are a trendsetter. For the current season the renowned label sets the standards with its collection for 2016. The CE2659 is available in the Edel-Optics online shop in other styles from Chloé collections 2015 and 2016.
The designer appeals to women who feel at home in the big cities. Mr. Right or not - the priority is to be en vogue in 2016. Full rim frames completely enclose the lenses within the frame. For a convinced eyeglass wearer, any other frame style is ruled out, with very few exceptions. round eyeglasses are extremely prestigious. A fact John Lennon was aware of, being rarely seen without them. Harry Potter would also not be the same without his characteristic eyeglasses. Due to its shape the CE2659 will be a long-time companion, because round eyeglasses never really go out of fashion. The colour red stands for love, but is also an eye-catching fashion statement. The colour ennobles your style and attracts attention - figuratively speaking and self-evident only for your "audience".
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text:
- {{matchup.date.fullday}}{{matchup.date.month + ' ' + matchup.date.day}}
UNC Asheville vs Virginia Intermont Final: 83-58 - Recap
UNC Asheville blew out Virginia Intermont on Friday, running up the score in an 83-58 win at Kimmel Arena.
UNC Asheville got 22 points from Andrew Rowsey and another 13 points from D.J. Cunningham. Jaron Lane contributed 4 assists, while D.J. Cunningham cleaned the glass with 14 rebounds.
Virginia Intermont got a 28-point performance from Joshua Hart, who was supported by 13 points from J.D. Tucker. Kris Harris had 3 assists in a losing cause for Virginia Intermont, while J.D. Tucker grabbed 6 rebounds.
UNC Asheville hit the break leading 29-23 before taking care of business in the second half.
UNC Asheville won the battle of the boards, outrebounding their counterparts 61-27.


text:
What is the poetic devices for the poem "If" by Rudyard Kipling?Please attach it with an example, thank you.
1 Answer | Add Yours
The three most common poetic devices that are presented in this poem are repetition, personification, and alliteration. Repetition is the repeated use of a word or phrase for a certain effect. In this poem, Kipling uses repetition with the word "you" throughout the entire poem to emphasize how important "you" is.
Personification is when a non human thing is given human qualities; there are three examples of personification here. The first is in Stanza 2, when he states,
"…make dreams your master…" (line 9)
The second example of personification also occurs in Stanza 2 when Kipling says,
"If you can meet with triumph and disaster
And treat those two imposters just the same." (lines 11 – 12)
The final example of personification is in Stanza 3, when Kipling states,
"If you can force your heart and nerve and sinew
To serve your turn long after they are gone" (lines 21 – 22)
Finally, alliteration is the repetition of the consonant sound at the beginning of consecutive words. There are at least two examples of this. The first occurs in stanza 2:
"with wornout tools" (line 16)
And the second example is from stanza 4 and says:
"sixty seconds" (line 30)
We've answered 333,670 questions. We can answer yours, too.Ask a question


text:
Opened 3 years ago
Closed 3 years ago
Last modified 3 years ago
#20142 closed Bug (fixed)
Transaction methods are not re-enabled when fixture loading fails
|Reported by:
|Owned by:
|Testing framework
|Triage Stage:
|Has patch:
|Needs documentation:
|Needs tests:
|Patch needs improvement:
|Easy pickings:
When a test case fails to properly load a fixture (integrity error or some other reason), the transaction methods (which are disabled before the test is run) are not re-enabled before the next test is run. This can cause other tests (especially transaction test cases) to fail.
I've created a simple django application to reproduce this. You just have to add the application to a Django project, and then run:
manage.py test myapp.Before myapp.Broken myapp.After
This command is used to enforce the test order that shows the bug. The Before and After tests are actually the same test that checks if transaction methods are enabled (as they should be for a transaction test case). The first one passes and the other one fails:
import django.db.transaction from django.test.testcases import TestCase from django.test.testcases import TransactionTestCase ORIGINAL = django.db.transaction.enter_transaction_management class TransactionCheckMixin(object): def test_transaction_method(self): self.assertEqual(ORIGINAL, django.db.transaction.enter_transaction_management) class Before(TransactionCheckMixin, TransactionTestCase): pass class Broken(TestCase): fixtures = ['broken_fixture'] def test_something(self): self.assertTrue(True) # will not happen class After(TransactionCheckMixin, TransactionTestCase): pass
The problem lies in the _fixture_setup method of the TestCase class:
class TestCase(TransactionTestCase): """ Does basically the same as TransactionTestCase, but surrounds every test with a transaction, monkey-patches the real transaction management routines to do nothing, and rollsback the test transaction at the end of the test. You have to use TransactionTestCase, if you need transaction management inside a test. """ def _fixture_setup(self): if not connections_support_transactions(): return super(TestCase, self)._fixture_setup() assert not self.reset_sequences, 'reset_sequences cannot be used on TestCase instances' for db_name in self._databases_names(): transaction.enter_transaction_management(using=db_name) transaction.managed(True, using=db_name) disable_transaction_methods() from django.contrib.sites.models import Site Site.objects.clear_cache() for db in self._databases_names(include_mirrors=False): if hasattr(self, 'fixtures'): call_command('loaddata', *self.fixtures, **{ 'verbosity': 0, 'commit': False, 'database': db, 'skip_validation': True, })
It should handle errors and call the _fixture_teardown method if there's an error.
Attachments (1)
Change History (5)
Changed 3 years ago by mlarente
Simple app that show the bug
comment:1 Changed 3 years ago by filias
- Needs documentation unset
- Needs tests unset
- Owner changed from nobody to filias
- Patch needs improvement unset
- Status changed from new to assigned
- Triage Stage changed from Unreviewed to Accepted
Confirmed the bug in django 1.5 and development version.
comment:2 Changed 3 years ago by filias
- Easy pickings set
- Has patch set
This fix does not contain tests as it is impossible to test it without mocking and a decision about using mocks in the project as yet not been made. This was discussed and agreed with Russel.
comment:3 Changed 3 years ago by Filipa Andrade <filipa.andrade@…>
- Resolution set to fixed
- Status changed from assigned to closed


text:
Just a quick one, what was the water quality like? Clean or a duck crap fest as was suggested last year?
I thought it was slightly cleaner than last year, but that may be due to the late spring and the goslings were younger/smaller and poo slightly less.
I have done a race report and posted it on the training for a shorter tri thread; that is where I normally post. It is long so you are warned. I will post here if wanted but as I had my first ever DNF it is a bit of a downer.
I am writing mine, but I've been down in Bristol, now back home and trying to get a job application sorted and get ready to go to Berlin for the Velothon.
It'll arrive shortly (maybe even tomorrow before I go)
On nights Matt?
Steady, post it here please. I was sooooooo gutted for you when I was you in the hall.
I will try to keep the story brief, but you are warned to have a cup of tea at hand, I do go on a bit.
I arrived lunchtime Saturday and registered, went to the 1pm briefing and met up with Saffers, Southern Snail, Mathschick, Matt and Chris. Nice to put faces to names now, I think we all wibbled together. Went back to campsite and put tent up, met up with Little Sister who was also camping and discovered where she would be marshalling.
Race morning, awake at 4am to lovely dawn chorus until the crows/rooks joined in. Took my tablets and started to faff. Porridge and mug of tea for breakfast with my mates who thoughtfully had boiled water up the night before and put into some flasks, it makes for lots less hassle. Took the bike and bits to transition, back to the tent to get into wetsuit. Here my tall tent came into its own and we managed to get 3 people into wetsuits inside where it was slightly warmer.
Went over to the start, as I went forward to the pontoon I caught my middle toe on the left foot on the leg of a barrier. Ouch, sod it and other words came out. No time to complain as we were herded into the water. Managed a bit of a warm up and went back towards the back of the first bay, up to my neck in water, standing around, so saved a bit of energy. 5 second countdown and then off, bit of biffing and bashing, but no worse than last year in the full Outlaw. Settled down and did my swim.
Swim time 50.37
Had a small struggle to get feet out of wetsuit but mainly because my hands were so cold, looked down at feet and just saw 2 slabs of white, no feeling. Slurped some drink, then changed into pirate cycle top, arm warmers, gilet, then as about to put socks and shoes on, dozy girl nearby asks politely if I could help her get her arms free from her wetsuit. Being a nice person I stopped and helped but did give out an exasperated sigh when she had the cheek to ask to be helped out of the 2nd arm. So socks and shoes on, grab bike and go.
Time 12.11
Bike ride around the lake, marshals on the far side did a grand job shooing geese off the road, coming back down 2 families of geese parted like the Red Sea for me. Saw Little Sister and I was off out on the roads. Made sure I drank regularly, took on gels, I enjoyed the closed road bits. I knew I was always going to be tight to the cut off time for the bike, so kept aero as much as I could and kept the cadence up. Each time I tried to change gear into what I knew I should be in the cadence dropped rapidly, so settled down to loads of spinning. I have a Garmin but with my contact lenses in I cannot read it so had to go by perceived effort.
I will admit to having to walk about 150m up the final bit of Oxton bank. I stopped at the feed station to ensure I got drink and another gel. Set off again, there were still a few riders in front and the occasional one overtaking me, no chance of drafting whatsoever. On the approach to the roundabout to start the southern loop a motorbike BTF official waved me down, oh no what violation have I done now.
No, the bad news was I was going to miss the cut off and although before the 12 noon deadline there was no way I was going to make it. He was an incredibly kind man, who had a middle aged, menopausal woman cry in front of him. At least where he stopped me meant it was a short 5 to 6 miles back to HPP following the race route. I had the option to continue knowing I would not be able to run, but there would be no marshals out and no back up. The thought at that point was I do not want to get lost in Nottinghamshire, I want to go back and bawl my eyes out, I had never felt so deflated in my life.
So shortcut back onto race route, oh how horrible that last bit is back past the posh house with the lousy surface. Back into HPP and did a spectacular dismount. I failed to dismount and the emotions got to me, so I just dropped the bike to the ground and stepped away. I made a comment to the marshal that there is more than one way to get off, just mine is not elegant. Racked the bike whilst controlling the tears, trying to work out what to do next.
This is when I discovered that my toes were painful and throbbing. I put my running shoes on and hobbled out, back to the campsite, feeling a total failure.
Sitting in the porch of the tent, I took my socks and shoes off to see a purple swollen middle toe, which was why I could not get enough power through the pedals. Painkillers taken, I phoned my husband at home and cried down the phone to him. After a while I realised I still had my timing chip on, but decided to have a shower, then go back and return it.
Back into race finish area, found an official to hand timing chip to, told that I can still have meal in food hall, so off I went, surrounded by masses of people in finishers T shirts with medals around their necks.
Pirates really are a great bunch of people, thanks to The Engineer; he was the first pirate I spoke to, sorry for interrupting your eating. Saffers gave me a hug and a talking to, thanks. Sorry to all for being a near blubbering wreck, but I guess you all know how much is put into preparing for the day. I went back to the campsite as I could not bear to be around all the happy faces, so I started getting packed up ready to go home. My friend Steve finished first out of the 3 of us, he was gob smacked to see me at the tent. His partner, known as Marmite Lady due to her cycle top finished a while later; she already knew that I had dropped out as she had not seen me on the run.
It was a long and painful drive down the M1 every time I changed gear, with horrible delays between 4 or 5 junctions and the traffic crawling along.
I took Monday off work as originally planned, and have had time to come to terms with what happened and why. Once I downloaded the Garmin, I was quite pleased to see that I had managed to do an average of 13mph, covering 33 + 5 miles out of the 56 with a badly bruised toe.
The extreme emotions were a combination of the day itself, my first ever DNF (I have run for 29 years and done Tri for 5 years) and being a woman of a certain age.
There will be a next time, it is being planned, but for now I will recover and do some shorter stuff, with some sportives in September.
Feel for you Steady, we had a chat to Marmite Lady after she finished and she said you are her neighbour, sorry we never got a chance to cheer you on on the run course
Thanks SA, I couldn't do your DIY one as it was Mr Steady's birthday. One day we will meet.
Nice report Steady, so sorry you didn't finish but I look forward to seeing you back there..... You have unfinished business
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Dr. Martens
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i have completed my graduation in nov 2012 an have worked for just for 2 months at sami labs and now am planing to start a drug distribution business in bangalore .
i have a little knowledge about it, i want to know more about the buisness like what will the investment , copetators ,legal formalities etc.........
kindly help me
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These are some of the stunning examples of the colourful images im a huge fan of colors and colorful images and these are some of the stunning examples of it .. Colors do effect our lives and they do color our lives ..
Color your life …
Paramount Pictures has now debuted the first Rings trailer and poster, Directed by F. Javier Gutierrez from a script by David Loucka, Jacob Estes and Akiva Goldsman. In Rings, a young woman becomes worried...
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5 Ways Batman: The Killing Joke Failed Audiences
15 Most Beautiful Female Athletes of the Rio Olympics 2016
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Definition of sanitarian in English:
nounchiefly archaic
An official responsible for public health or a person in favour of public health reform.
Example sentences
- Shane works for the Worth County Conservation Board as a conservation technician and also for the Worth County Public Health as the county sanitarian.
- Now, the working groups are represented by manufacturers, regulatory officials, sanitarians and end users,' he explains.
- It is best to discuss results with a sanitarian from the local health department, since he or she will have these established levels available and can help you interpret your results.
Words that rhyme with sanitarianagrarian, antiquarian, apiarian, Aquarian, Arian, Aryan, authoritarian, barbarian, Bavarian, Bulgarian, Caesarean (US Cesarean), centenarian, communitarian, contrarian, Darien, disciplinarian, egalitarian, equalitarian, establishmentarian, fruitarian, Gibraltarian, grammarian, Hanoverian, humanitarian, Hungarian, latitudinarian, libertarian, librarian, majoritarian, millenarian, necessarian, necessitarian, nonagenarian, octogenarian, ovarian, Parian, parliamentarian, planarian, predestinarian, prelapsarian, proletarian, quadragenarian, quinquagenarian, quodlibetarian, Rastafarian, riparian, rosarian, Rotarian, sabbatarian, Sagittarian, Sauveterrian, sectarian, seminarian, septuagenarian, sexagenarian, topiarian, totalitarian, Trinitarian, ubiquitarian, Unitarian, utilitarian, valetudinarian, vegetarian, veterinarian, vulgarian
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Billed as a 'Next Generation Interactive Whiteboard with intuitive pen and touch' —
when you use it for yourself or at least see this video—you'll see why. The pen and touch capability really makes learning with it look like great fun. It works with Windows 7 and integrates with Promethean's ActivClassroom solutions and content. There are math tools, sounds, templates, and gesture touch applications available as teachers and students discover a natural way of interacting with technology. It's neat how images can be easily moved, scaled and rotated with finger touch and a pen. Reminds me a little of Spielberg's Minority Report technology, where a lot of dramatic arm-waving, shuffling of images were part of the future. Have a look at the video and see what I mean. The video is wordless, as that would require translation (Promethean sells worldwide); funny how technology can be so global and so universal. The piano music is nice, too.
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- Report: #845246
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Complaint Review: slotnuts casino online
slotnuts casino onlineInternet United States of America
slotnuts casino online ,slotnuts.com accounting keeps saying i was all ready approve on payout Internet, Internet
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nope know had called me on the e-mails-also i chat on live support and they realy don't say much just gives me the round around.im all ready tired of all this..this is not fair to me..,i try ed to get the i pad number on the wire transfer so my bank can research it on this end they wont even give that to me, they state the people that does the accounts ant in tiled to that information ,but in there world they keep saying to ..rest a sure you will get your payout..im tired of hearing that..my id there is debbie49 my email is(((REDACTED))) @sbcglobal.net
CLICK here to see why Rip-off Report, as a matter of policy, deleted either a phone number, link or e-mail address from this Report.
This report was posted on Ripoff Report on 02/27/2012 09:31 AM and is a permanent record located here: http://www.ripoffreport.com/r/slotnuts-casino-online/internet/slotnuts-casino-online-slotnutscom-accounting-keeps-saying-i-was-all-ready-approve-on-pa-845246. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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"The baby brunch invite is very beautiful. The paper in thick card stock and the envenlopes are a very good quality. It was fairly easy to customize my order. I thought I had put in some wrong info. I e-mailed them, and they were able to fix it for me."
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A new exhibit at The California Museum in Sacramento features arts and crafts from the internment camps where Japanese-Americans were forced to relocated during World War II.
Many of the items on display in "The Art of Gaman: Arts & Crafts from the Japanese American Internment Camps, 1942-1946" were made by farmers, shopkeepers and housewives from the San Joaquin County including Lodi and Stockton.
"This is party due to the fact that I grew up in Lodi and when I started gathering objects for my book and exhibition, I asked friends and relatives in this area first," said Delphine Hirasuna, author of 2005 book "The Art of Gaman" and a guest curator of the exhibit opening Sunday.
The artifacts made by internees included intricately carved birds, watercolor paintings and teapots.
Location, ST | website.com


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Omar Figueroa To Vacate 135 Title, Moving Up to 140
By Jake Donovan
Omar Figueroa decided to save his best lightweight performance for his last. The unbeaten Texan informed the World Boxing Council (WBC) on Tuesday of his plans to vacate his title and campaign in the 140 lb. division, where he plans to pursue a showdown with lineal champ Danny Garcia.
The news comes on the heels of Figueroa's thrilling 9th round knockout of Mexico's Daniel Estrada this past weekend at the StubHub Center in Carson, California. The bout was by far the most thrilling of the evening, also serving as the opening leg of a Showtime-televised tripleheader. Figueroa fended off a stiff challenge from Estrada, overcoming a cut suffered in round eight to stop his mandatory challenger in the very next round.
Whispers swirled leading into the fight of Figueroa (24-0-1, 18KOs) struggling to make the lightweight limit, with the belief that he stuck around long enough to capture a title eventually moving up in weight. The all-action rising star first gained notoriety with a Jan. '12 knockout win over Michael Perez, unbeaten at the time and the better known commodity who was being groomed for stardom.
The win instead propelled Figueroa towards contention, although the journey was stalled as adviser Al Haymon preferred an active itinerary against journeyman-level competition while seeking the best opportunity down the road. That moment came last summer, when an interim title was made available for his eventual 12-round thriller with Nihito Arakawa in one of the best fights of 2013.
Figueroa was granted full status of the belt after it was officially vacated by Adrien Broner. His lone other defense prior to Saturday came this past April, struggling to a 12-round decision over Jerry Belmontes in his first fight back following a lengthy hiatus while allowing his oft-damaged hands to fully heal.
While it is unlikely that he lands a showdown with Garcia, a title fight figures to be in Figueroa's future once he formally moves up to super lightweight. Garcia has been ordered to defend one of his belts versus mandatory challenger Viktor Postol, while also linked to a possible showdown with Lamont Peterson, who also owns a title in the 140 lb. division.
All of this comes while there remains the strong possibility that Garcia eventually outgrows the division altogether. His last fight was a 142 lb. catchweight versus overmatched Rod Salka, and has hinted at a possible Peterson showdown taking place outside the weight class before officially moving up to welterweight.
Whatever the case, Figueroa will somehow land in the thick of things once he's fully ready to make the move five pounds north, which is expected to come once he resumes training after allowing his cut to fully heal.
The greatest benefactor of the announcement will be former two-division titlist Jorge Linares, who was due to face the winner of Saturday's lightweight title fight. Linares appeared on the undercard, knocking out journeyman Ira Terry in two rounds. His next fight will most likely be for the vacant lightweight title, against the next highest rated lightweight contender once the next set of rankings are released by the WBC.
Jake Donovan is the Managing Editor of BoxingScene.com, as well as the Records Keeper for the Transnational Boxing Ratings Board and a member of Boxing Writers Association of America. Twitter: @JakeNDaBox
[QUOTE=Vicious.;14873889]I dislike haymon fighters as much as the next guy but nobody is ducking linares. Fig would ko his ass[/QUOTE]Linares with his teeny, tiny, pointed chin gets flattened every time he fights a guy with a punch.Comment by Vicious. on 08-20-2014
[QUOTE=manoamano;14873058]What could we expect from another haymo fighter. Vacates the title when Linares was after him. Dude has enough body fat and can't make weight cause he eats a lot! How many defenses does he have and against what level…Comment by manoamano on 08-20-2014
What could we expect from another haymo fighter. Vacates the title when Linares was after him. Dude has enough body fat and can't make weight cause he eats a lot! How many defenses does he have and against what level…Comment by richardt on 08-20-2014
[QUOTE=HanzGruber;14871804]They're not guys that should be making him look like that though lol. Hell get beat like demarco vs broner. That's how bad his defense is[/QUOTE]Unless or until then, he will bring it every time in the ring. It's not…Comment by nycsmooth on 08-20-2014
this is a fairly weak division, & guessing Estrada falls down to 4 or 5, the guy next after Linares, who should move 2 #1 is, & I can't believe in last months wbc ranking I saw, ranked #3 (…Post a Comment - View More User Comments (27)


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What You Can Do To Get Rid Of Fleas From Cats It is always great to learn about the best flea medicines for cats and other pets so you can also make sure that you are protected in terms of your health. These are among the greatest concerns that are faced by pet owners because the many various flea treatments and the best flea medicines for cats have had several reactions for their specific body types. It is important that you can ensure that whichever treatments that you let your cats to take, there can be no side effects and the desired results are achieved. When you are looking for the best flea medicines for cats, be sure that you can look for the right ones through having the products tested before you can buy them out from the stores. It is important to note that there are several pet veterinarians who have shared their opinion about the effects of the best flea medicines for cats, but they also noted how these treatments can always be used for the desired outcomes such as regular check-ups every once in a while. There are experts who advised these pet owners that they have to make sure that these medicines, classified on their strength and their ingredients, can be able to provide them the best results. It is important to consider that much of these best flea medicines for cats are in fluid form and they are poured over the skins of the cats, particular in between the shoulder blades where these cats cannot lick them off. It is just right to see and hear about questions that are asked by the cat owners and seeing their efficiency when used. Many of these factors can consider the demand for the best flea shampoo for cats as treatments for these infections and several people are looking for more eco-friendly ways to apply them so minimal risks are posed.
The Beginners Guide To Treatments (Getting Started 101)
Since many of these cats are experiencing some effects from the fleas, it is imperative for owners to consider the flea treatments for cats and they should always think about some factors before buying these things. There are several treatments that contain active formulas that can be able to treat these allergies, but be aware of the side effects. If you find that these products can cause some risky effects for your pets, worry not because there are alternatives that the market renders as available for you. Worry not about these flea treatments for cats because you can always avail of medicines that can work within a day or even shorter.The Beginners Guide To Treatments (Chapter 1)
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- Report: #427314
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Complaint Review: Whirlpool
Whirlpool553 Benson Rd Benton Harbor, Michigan U.S.A.
Whirlpool Corporation Defective Whirlpool Front Loader Washer Benton Harbor Michigan
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First socks were getting caught in the sud(sunk pump). I had someone to come out to service the washer. He suggested I put all socks in bag. I did that. Then other items started getting caught in the sud. It is almost as if everything you wash have to be put in a bag in order for it not to get caught in the pump. It became nerve racking for me to worry about everything I put in the washer may get stuck. It happened again, with a shirt, then again. Then the hose busrt. Someone had to come out and fix the hose.
Now, the washer is not working because water is coming out of area where I put the detergent each time I try to wash. I will never buy a whirlpool product again. I contacted whirlpool a couple of months ago. They defended their product. My warrenty expired 9-2008. Whirlpool agreed to send someone out to fix the hose only because I told them I would be going to the better business bureau in which I still am. That was 1 month ago. Now the washer is not working again because water is coming out the area where the detergent go. I was sold a lemon. I am very upset. Besides the BBB is there anything I can do to get a new Washer?????
Florissant, Missouri
This report was posted on Ripoff Report on 02/22/2009 11:53 PM and is a permanent record located here: http://www.ripoffreport.com/r/Whirlpool/Benton-Harbor-Michigan-49022/Whirlpool-Corporation-Defective-Whirlpool-Front-Loader-Washer-Benton-Harbor-Michigan-427314. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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Turn your event gray with our Flat Paper.
Cover your walls or make fabulous signs with this Gray Flat Paper. Quickly and easily decorate large or unsightly areas with this great paper. Each roll measures 4 feet x 50 feet and is white on the back side.
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Bought this to use for pictures to cover up floor tiles - not made for walking on - tears fairly easy - also put on sides of props - tears if doesn't have something solid behind it. Wasn't expensive though


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How can you tell if LO does not like the nanny? When we are home or if anyone else watches him, he keeps his schedule. Sleeps and eats when he is supposed to. When the nanny watches him he is crying all the time and not sleeping or finishing his bottle. Could it be the nanny? Or separation anxiety? I just went back to work last week, but the nanny has been with us for a month now. My LO is 5 months. Any idea what it could be?
How do you know if LO does not like the nanny?
Comments (3)
I don't think at 4-5 month age, your child gets much seperation anxiety. It mostly starts around age 1.
My best guess will be your nanny is not doing something right. Have you worked with her closely and watch her doing what she does when she first working with you? I would take a couple of days off and just observe at home to see what she has been doing "wrong". Baby loves routines. And if she does something different from what he is used to, then that could be a problem. GL!
Theoretically separation anxiety starts around 9 months, but in my case, when I went back to work and my LO was 4 months old, I had a similar issue.
Even though my mom was taking care of my daughter, and my daughter loves her, she was still getting upset when I was leaving in the morning.
So, it is possible that it is a combination of both - the nanny not doing something right, plus your LO misses you.
If this is a new thing then yes, maybe LO is just upset your gone. How does the nanny feel about it, could you maybe buy a cheap nanny cam (and tell her of course) in an attempt to work out what she is doing differently? It might not be anything bad but hat she does things differently and it's not working for your LO.