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These studies were structured with the aim of utilizing emerging technologies in two-dimensional (2D) gel electrophoresis and mass spectrometry to evaluate protein expression changes associated with type 1 diabetes. We reasoned that a broad examination of diabetic tissues at the protein level might open up novel avenues of investigation of the metabolic and signaling pathways that are adversely affected in type 1 diabetes. This study compared the protein expression of the liver, heart, and skeletal muscle of diabetes-prone rats and matched control rats by semiquantitative liquid chromatography-mass spectrometry and differential in-gel 2D gel electrophoresis. Differential expression of 341 proteins in liver, 43 in heart, and 9 (2D gel only) in skeletal muscle was detected. These data were assembled into the relevant metabolic pathways affected primarily in liver. Multiple covalent modifications were also apparent in 2D gel analysis. Several new hypotheses were generated by these data, including mechanisms of net cytosolic protein oxidation, formaldehyde generation by the methionine cycle, and inhibition of carbon substrate oxidation via reduction in citrate synthase and short-chain acyl-CoA dehydrogenase.
- metabolism
- acyl carnitine
- citrate synthase
- methionine
- liver
- heart
- skeletal muscle
- mass spectroscopy
- two-dimensional gel electrophoresis
although extensively studied, type 1 diabetes remains a major health concern, and much of the pathogenesis still eludes understanding. In the Diabetes Control and Complications Trial, the most strongly regulated subjects still developed complications despite aggressive intervention (35). The fundamental pathology of type 1 diabetes is the loss of pancreatic β-cell function and the related release of insulin in the regulation of overall metabolic functions of the body. The complications of type 1 diabetes include dramatically altered energy metabolism and glucose handling, protein oxidation, vascular disease, heart disease, retinopathy, neuropathies, nephropathy, and susceptibility to infections. These results showed that the pathology of diabetes exceeds the simple maintenance of plasma glucose and insulin levels. A more global assessment of the diabetic state may provide useful insight. These studies sought to perform a comprehensive screen of the proteomic changes associated with the type 1 diabetic state, utilizing emerging technology in high-throughput semiquantitative proteomics and two-dimensional (2D) gel electrophoresis to detect protein expression differences as well as possible covalent modifications inherent to the diabetic state.
Previous studies have revealed that many enzymes and proteins are under transitional control by insulin (45) that would predictably alter the content of these proteins with the reduction of insulin in type 1 diabetes. The major transcription factors involved in insulin action are adipocyte differentiation and determination factor (ADD)-1/sterol regulatory element binding protein (SREBP)-1/peroxisome proliferator-activated receptor (PPAR)γ, involved in fatty acid and triglyceride metabolism as well as other aspects of intermediary metabolism (61). Several recent screens of gene expression are available in various models of diabetes consistent with a key role of insulin in the gene expression profiles associated with energy metabolism as well as many other metabolic as well as signaling pathways (15, 64). We reasoned that a proteomic screen might provide insight into the complex metabolic and regulatory pathways associated with type 1 diabetes by directly monitoring the protein levels with recently developed differential quantitative proteomic approaches.
Since we chose to examine multiple organs in these studies, we selected an animal model with a matched control. The most common model used in diabetes research is streptozotocin or alloxan treatment, which results in β-cell death, insulin insufficiency, and hence diabetes (5, 53). Although these methods produce animals that model diabetes, streptozotocin and alloxan are oxidative stressors and the secondary oxidative stress effects can confound diabetes in these animals. Because mitochondrial proteins were of interest and these are particularly susceptible to oxidative stress, we decided not to use alloxan and streptozotocin. The use of knockout mouse models is problematic because the mutations that are introduced may induce hyperglycemia by mechanisms that do not normally occur in diabetes. Mice also yield so little tissue that analysis via 2D gel and mass spectrometry without pooling samples becomes problematic. The two rat models considered were the Zucker diabetic fatty/type 2 diabetes (ZDF) model (genetic models) and the Bio-Breeding diabetes-prone (BB-DP) spontaneous type 1 diabetes model (biomedical research models). We chose to avoid the confounding variables associated with obesity in conjunction with diabetes in the ZDF rat. On the basis of these numerous considerations, we selected the BB-DP model. The BB-DP strain is maintained commercially along with a matched-background diabetes-resistant strain (BB-DR) and follows a course similar to human type 1 diabetes, with 96% of the rats becoming diabetic by 120 days. The course is predictable and is autoimmune, similar to human diabetes. The lipid profiles of the rats are similar to the human condition (39, 49).
We used 2D gel electrophoresis to qualitatively screen for protein content, hydrolysis (decreases in molecular weight), and posttranslational modifications [as demonstrated by isoelectric focusing variants (IEV)]. IEV can reflect several different posttranslational modifications from phosphorylation, oxidation, ADP-ribosylation, or even selective protein cleavage. Any modification of the protein that would change its surface charge will result in an IEV. Thus this measure must be interpreted with caution. Liquid chromatography-mass spectrometry (LC-MS) was used for more extensive determination of protein content differences associated with the diabetic state because LC-MS has a broader range than standard 2D gel electrophoresis. 2D gel analysis was performed on diabetic and control heart, liver, and muscle. LC-MS analysis was performed on liver and heart, where the initial 2D gel studies demonstrated the largest protein content changes with diabetes. These data were then compiled into a format that permitted functional evaluation of the protein alterations associated with defined metabolic pathways. In addition to confirming some of the known changes associated with diabetes, this approach provided evidence for novel alterations that merit further studies by conventional experimental approaches.
Tissue harvest.
Urine glucose levels were measured daily after the BB-DP rats were ∼7 wk old. Urine glucose-positive animals were subjected to blood glucose measurements via tail blood and glucose analyzer determination. To prevent the development of insulin antibodies and potentially confounding data, rats were not treated with insulin. Animals were allowed ad libitum standard chow until the day of tissue harvest. Blood glucose was measured twice daily until blood glucose was ≥500 mg/dl and/or the rats demonstrated significant ketonuria. Fluid therapy was administered if necessary to maintain health. Within 30 h of ≥500 mg/dl blood glucose and/or significant ketonuria, rats were euthanized. This usually occurred when the rats were ∼17 wk old. All necropsies were performed at noon to minimize protein expression differences related to circadian rhythm. Acutely diabetic BB-DP and age-matched (±3 days) BB-DR rats were fasted from 7:00 AM until euthanasia on the day of collection. Euthanasia was by decapitation. The guillotine was thoroughly washed and the area of decapitation scrupulously cleaned to prevent stressing the rats en route to euthanasia. Animals were kept in a room separate from the site of decapitation and transported to the guillotine immediately before euthanasia. The whole heart was removed and washed with ice-cold saline in three consecutive dishes. Two pieces from the left lobe of the liver were removed (∼5 g each) and placed briefly into three consecutive petri dishes with ice-cold saline to chill and remove blood. Approximately 5 g of skeletal muscle from the upper leg composed primarily of the biceps femoris, adductor magnus, and gracilis was collected and washed with ice-cold saline. The tissues were immediately placed in cryovials and flash frozen with liquid nitrogen. Total time from decapitation until all tissues were frozen never exceeded 2 min.
Six sets of male rats were selected for proteomics experiments, all of which were killed at blood glucose >500 mg/dl. All rats harvested were within 9 days of age of each other and control animals. All procedures performed were in accordance with the guidelines described in the Animal Care and Welfare Act (7 U.S.C. 2142 §13) and under a protocol (H-0069) approved by the National Heart, Lung, and Blood Institute Animal Care and Use Committee.
Frozen tissue was pulverized in liquid nitrogen with a bio-pulverizer apparatus (59014H Biospec Products) with at least 30 hammer strikes or until completely powdered. Samples were suspended in standard lysis buffer, 7 M urea, 2 M thiourea, and 4% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). The tissue powder was suspended in 8 ml lysis buffer/g tissue powder for 45 min at room 20°C with frequent vortexing. Fifteen millimolar Tris buffer, pH 8.5, was added to the lysis buffer for 2D gel experiments. Samples were centrifuged at 100,000 g for 35 min. Lipid phase and pellets were discarded. The rest of the tissue extract was preserved for gel studies and analysis by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) MS.
2D gel electrophoresis and protein labeling.
Concentrations of purified lysates from tissue samples of diabetic and control animals were determined with the GE Biosciences-Amersham 2D Quant-Kit and an in-house spectrophotometer per the manufacturer's instructions. Fifty micrograms of control and diabetic protein was labeled with differential in-gel 2D electrophoresis (DIGE) Cy3 and Cy5 dyes, respectively. A standard containing 25 μg of each sample was labeled with Cy2 per the Amersham DIGE protocol (200 pmol/μl labeling). The reaction was then quenched with 10 mM lysine for 10 min. Labeled Cy2-, Cy3- and Cy5-treated samples were combined with 8 μl of DeStreak (GE Biosciences-Amersham) and 4 μl of pH 3–10 ampholytes (GE Biosciences-Amersham). Solution volume was adjusted to 440 μl with the necessary volume of rehydration solution. Samples were loaded onto 24-cm Immobiline DryStrip pH 3–10 (GE Biosciences-Amersham). Isoelectric focusing was achieved by active rehydration for 11 h at 30 V followed by stepwise application of 500 V, 1,000 V, and 8,000 V for a total of ∼70,000 Vh (Ettan IPG Phor, GE Biosciences-Amersham). Strips were equilibrated in equilibration solution (50 mM Tris-Cl, 6 M urea, 30% glycerol, 2% SDS) with 0.5% DTT for 12 min with agitation and then in equilibration solution with 4.5% iodoacetamide for 12 min with agitation. Strips were loaded onto 10–15% SDS-PAGE acrylamide gradient gels (Jule) and sealed with 0.5% agarose. The second-dimension gels were run at 13°C for 30 min at a constant 50 V and then for ∼18–21 h at 105 V constant voltage. Gels were rinsed with purified water and then scanned on a Typhoon 9400 scanner (GE Biosciences-Amersham) at 100-μm resolution. Cy2 gels were scanned at 540 Bp40 on a blue2 (488 nm) laser, Cy3 gels were scanned at 580 Bp30 with a green (532 nm) laser, and Cy5 gels were scanned at 670 Bp30 with a red (633 nm) laser. Gels utilized for MS analysis were run with 500 μg of protein. Plates were separated, and gels were fixed overnight in 30% methanol and 10% glacial acetic acid. Gels were then stained overnight in Sypro Ruby dye (Molecular Probes).
Differential gel presentation was created within a custom-written image processing program (IDL, RSI) to correct for scanning gain/detector alterations for the different wavelengths. Since large differences in protein contents were observed in some of these studies, global corrections were found to be inadequate. For this procedure, a nonscaled color overlay of the Cy3 and Cy5 images was used to select a region of interest (ROI) for use in the scaling function. The ROI was selected based on an area of the gel in which there was high signal-to-noise difference in Cy3 and Cy5 with minimal background noise; this optimized the linear least squares approach using the scaling function. In general, the ROI represented <20% of the total image. A linear least squares analysis between the three channels collected was performed with the following equation: ICy3[Cy3(ROI)] + ICy5[Cy5(ROI)] = Cy2(ROI) (1).
The ICy3 and ICy5 coefficients were determined with a resident least squares fitting routine within the IDL software environment, minimizing the residuals. To create the final DIGE image, the entire Cy3 and Cy5 images were overlaid after multiplying by the appropriate coefficients. This approach was accurate over a fivefold range in signal amplitude in protein standard experiments. In general, ICy3 and ICy5 ranged from 0.75 to 1.25 in the experimental samples.
Spot picking and protein identification.
Spot identification was carried out with an Ettan Spot Handling Workstation (GE Biosciences-Amersham) that performs automated extraction and in-gel trypsin digestion of selected protein spots according to an Amersham protocol. Peptides derived from protein spots of the gel were analyzed with a mass spectrometer (4700 Proteomics Discovery System, Applied Biosystems) using MALDI-TOF and tandem mass spectrometry (MS/MS). At least two peptides were obtained for each protein with MS/MS. Proteins were identified from the acquired spectra with the MASCOT database search function.
Sample preparation for mass spectrometry.
Liver or heart tissue was powdered on dry ice as described above. LC-MS was preformed and statistical analysis was done as described previously (27). Protein extraction and MS were performed as described previously (27, 33).
There was some concern that the magnitude of protein level changes might be underestimated. In previous studies (32, 33) the ratio of proteins was underestimated as the difference in protein content increased because of the effect of weak background signals preventing the denominator from reaching true zero. To evaluate this, we conducted a trial using five known proteins [APE-1, serine esterase, pyruvate dehydrogenase (PDH), arylamine-N-acetyltransferase, and alcohol dehydrogenase (ADH)] at three different concentrations within a mitochondrial protein background. These standards demonstrated consistent agreement in direction, but the method consistently underestimated the magnitude of difference. That is, the standards revealed a proper qualitative increase in measured amplitude with concentration, but all underestimated the relative increase in protein content. When measured protein ratio between samples was plotted versus the known amount of protein, the slope of the line was 0.5 (y = 0.38 + 0.5x; R = 0.86) rather than 1 for a perfect quantitative analysis. These data are presented in Supplemental Fig. S1.1 These data demonstrate that the relative concentrations of proteins are qualitatively correct; however, the precise ratio of proteins between samples is underestimated by roughly a factor of 2 in the mitochondrial protein environment.
DIGE was applied in control and diabetic heart, skeletal muscle, and liver protein homogenates as an initial screen. Heart and skeletal muscle demonstrated considerably less variation between the diabetic and control tissues compared with liver (Fig. 1). The protein content differences are presented in Supplemental Table S1 along with any changes detected by IEV (labeled as shift) with diabetes. The liver revealed numerous changes in protein expression in diabetes and numerous posttranslational modifications reflected in changes in the generation of IEV (Fig. 1). The DIGE approach provided a visual overview of the protein differences and detected some proteins not found in the MS studies with our conservative criterion. The major advantage of the DIGE approach was the screening of posttranslational modifications based on IEV generation. There were consistent alterations in the IEV patterns of aldehyde dehydrogenase 1 (liver), urocanase (liver), fumarase (skeletal muscle, heart, liver), HSP70 (liver), enolase (skeletal muscle, heart), tripartite motif containing protein 72 (heart, skeletal muscle), and glycogen phosphorylase (heart).
On examination of the DIGE data, many spots were non-Gaussian, with highly variable background levels making absolute quantitation difficult, especially with the low-abundance proteins, where "bleed over" from more abundant proteins was problematic. In addition, any protein greater than 100 kDa or with an isoelectric point outside pH 3–10 was not detected by these methods. Because of these limitations and the large number of changes that were identified, we chose to rely on a semiquantitative LC-MS approach mostly focused on the liver: 8,735 liver peptides were identified and quantified, resulting in 4,686 protein identifications. Of these, 555 proteins were identified with high confidence: 365 proteins had significant changes with P values of <0.05, and 194 proteins were >20% different, including 65 changes not previously reported in the diabetes literature to our knowledge. In the heart, there were several protein changes that were of interest. Pyruvate dehydrogenase kinase isoform 4 (PDHK4) was strongly upregulated, as previously demonstrated (67), in addition to several proteins that have not previously been associated with diabetes. We confirmed the increase in PDHK4 and other PDHK isoforms by Western blotting on the crude tissue homogenates (Supplemental Fig. S2).
To better understand the functional implications of the protein changes with diabetes, the liver data were used to generate graphic canonical models of represented metabolic pathways. Reasonable coverage of the proteins associated with glucose, fatty acid, tricarboxylic acid, urea, reactive oxygen species (ROS), and oxidative phosphorylation was seen in this screen (Figs. 2–4). We utilized a canonical approach that essentially assumes that the capacity of a given reaction step and metabolic pathway is proportional to the enzyme concentration. Naturally, this is a simplification because of the effects of posttranslational modifications, substrate/product/inhibitor interactions, and numerous allosteric effectors; however, it does provide a method of building hypotheses from this data set. What we sought to determine is whether the protein expression values would recapitulate the metabolic data that have been collected in diabetes and generate hypotheses about the metabolic pathway that have not previously been appreciated in studies that were more focused in their data collection methods.
Proteomics was used in this study to semiquantitate changes in protein expression patterns that might reveal metabolic changes in type 1 diabetes that are currently not appreciated. The intent was to obtain leads from this broad data collection method that could form the basis of hypotheses for future studies. In the diabetes model used, the most dramatic proteomic alterations were observed in the liver, with more subtle effects observed in skeletal muscle and heart. These data imply that the major adaptations occurring in this model of type 1 diabetes were occurring in the liver, with more modest changes occurring in the heart and skeletal muscle. These results are consistent with the known large alterations in liver metabolism associated with a lack of insulin mimicking systemic starvation. The general modifications to the proteome and likely function within the heart and skeletal muscle are relatively smaller.
In general, alterations in a given metabolic pathway were reflected by net changes in enzymes of the whole pathway rather than a just a few "rate-limiting" reactions, with the exception of the citric acid cycle and glycolysis. This is intuitive, as these pathways have multiple metabolic input and output nodes. This contributed to the large number of liver protein alterations observed (Fig. 1), since many proteins associated with each pathway were altered. This observation is consistent with the previous screen of the mitochondrial metabolic pathways between different tissues with different metabolic requirements that also reflected entire metabolic pathways alterations (32, 33).
For many of the proteins that were altered in the diabetic animal at levels >50%, we found excellent correlation with gene expression data from individual experiments looking at effects of insulin as well as screening studies in different diabetic animals (see Supplemental Table S2 and references). Since no gene expression screen has been conducted on the BB-DP rat, a direct comparison between the proteomic data and screens is difficult. It is interesting to note that many of the screen studies in diabetic models (13, 15, 64) did not detect gene expression changes in many genes that have been found to be altered in focused studies on similar diabetic models (for example, see Ref. 45). This observation suggests that the statistical power of these particular gene expression screens may not be approaching the sensitivity of the more selective studies or the present proteomic approach. It is also important to note that gene expression measurements are not dependent on the expressed protein concentration, only the generation of mRNA. Thus gene expression techniques detected production changes in low-concentration signaling proteins and transcription factors that were not detected in our protein screens. This discrepancy is likely due to the poor sensitivity of the MS detection in these complex samples.
To aid in the discussion of the predicted biochemical consequences of the protein changes detected, we have consolidated the data into several canonical pathways. The remaining discussion is focused on what the proteomics data suggest about these metabolic pathways.
Oxidative phosphorylation.
It is well established that the net oxygen consumption of the liver cell is increased in diabetes, consistent with an increase in ATP turnover secondary to increased metabolic work in the liver. For example, Exton and colleagues (18) demonstrated a 1.74-fold increase in oxygen consumption with diabetes, corresponding to a 2-fold increase in urea synthesis. Consistent with these observations, our proteomic data indicate that the capacity of the electron transport chain to produce ATP, and potentially therefore ROS, was upregulated in the liver at steps including complex 1, complex 3, complex 4, ANT1, ANT2, F0F1 complex, and the electron transport flavoprotein (ETF) (Fig. 2). These data are consistent with an increase in the ATP-generating capacity in the diabetic liver cell resulting from a general increase in the overall enzymatic content of this reaction sequence. The generalized increase in oxidative phosphorylation enzymes is consistent with an increase in mitochondria biosynthesis associated with the increase in PPARγ coactivator (PGC)-1α that occurs in the diabetic liver (25).
Citric acid cycle.
It has been demonstrated in the literature that the citric acid cycle is downregulated in diabetes (38). However, the specific modifications to the citric cycle enzymes have not been described in detail. According to the proteomic data collected in these studies, only two citric acid enzymes were downregulated in diabetes (Fig. 2): succinyl-CoA ligase and citrate synthase (CS). A posttranslational modification to fumarase was observed with an acid shift in its IEV, consistent with protein phosphorylation. The nature of this posttranslational modification of fumarase is unknown. There was an increase in both cytosolic (c) and mitochondrial (m) aspartate transaminase (AT). The latter two enzymes could decrease the mitochondrial matrix oxaloacetate by facilitating the export of oxaloacetate from the mitochondria for gluconeogenesis (discussed below). Together, these modifications could impede the citric acid cycle by both decreasing the entry of acetyl-CoA via CS and decreasing matrix oxaloacetate. CS levels have also been suggested to influence metabolic changes associated with aging (28, 57) and in caloric restriction (26). Finally, CS concentration is frequently used as a global measure of mitochondrial content/activity (37). However, as demonstrated in this study, CS is specifically downregulated in diabetes compared with enzymes involved in oxidative phosphorylation or fatty acid oxidation (FAO). Thus the reliance on CS concentration to quantitate mitochondrial content or as a standard in relation to mitochondrial function is tenuous.
Fatty acid oxidation.
Consistent with the previous documentation of an increase in FAO in the diabetic liver (20, 43), numerous enzymes associated with FAO were increased in the diabetic liver, including most of the acyl-CoA dehydrogenases, the three enzymes responsible for unsaturated fatty acid oxidation, carnitine palmitoyltransferase 2, the trifunctional enzyme complex (TFEC), and the previously mentioned ETF (Fig. 2). Thus the higher liver ATP generation capacity indicated by enzymes of oxidative phosphorylation is likely supported by FAO in diabetes. Interestingly, Krebs predicted that the acetyl-CoA generated via FAO is likely not entering the citric acid cycle but shunted to ketone bodies (20, 36), which has been supported by numerous subsequent studies that show that low matrix oxaloacetate inhibits citric acid flux under these conditions (17). Our data suggest that the enhanced diabetic ketone body formation from acetyl-CoA may be due in part to the downregulation of CS and the increased export of oxaloacetate from the matrix by the two isoforms of AT. Furthermore, a substantial change was apparent in the competition for acetyl-CoA between CS for citric acid cycle activity and hydroxymethylglutaryl-CoA (HMG-CoA) synthase for ketone body synthesis. As shown by our LC-MS standard curves, CS decreased by 60% while HMG-CoA synthase increased by >300% in diabetes, suggesting that ketone body formation would favorably compete for acetyl-CoA in these livers.
Short-chain acyl-CoA dehydrogenase (SCAD) was not upregulated in diabetes, despite the requirement that all FAOs must be processed through this enzyme for completion. The FAO pathway was upregulated with the exception of SCAD. This could potentially cause the buildup of short-chain fatty acyl-CoA, which in conjunction with an increase of carnitine acyltranslocase (lower-quality ID but significantly increased P value of 0.000025) suggests that short-chain acyl-carnitine may be released from the diabetic liver, allowing the recycling of CoA in the presence of decreased flux through SCAD. There is evidence that short-chain acyl-carnitines are increased in the serum of diabetic and fasted patients and that patients with diabetes can become carnitine depleted in liver tissue (12, 30). It is also of interest to note that the short-/branched-chain CoA dehydrogenase was upregulated in the heart, which would allow the handling of the increased short-chain acyl-carnitine released by the liver. To our knowledge, this is the first study that has demonstrated a differential regulation of acyl-CoA dehydrogenases, and it suggests a novel mechanism for the liver to deliver reducing equivalents and carnitine to the periphery via short-chain fatty acids.
A decrease in glycolysis and an increase in gluconeogenesis in diabetes are well documented (48). We observed a decrease in pyruvate kinase, one of the key enzymes in the control of glycolysis flux, enolase, and a single peptide identification of glucokinase (GK) (Fig. 3). A surprising decrease in glucokinase regulatory protein (GKRP) was measured in the diabetic animals in the DIGE studies. Since GKRP is generally believed to inhibit GK by sequestration in the nucleus, its downregulation is counterintuitive in a state in which glycolysis is suppressed. However, GKRP and GK contents have been directly correlated in GKRP knockout studies (24). Supporting the notion that GKRP is suppressed in diabetes, overexpression of GKRP in diabetic mice increases GK protein levels, increases GKRP activity, and improves glucose tolerance (59). The specific mechanisms related to the coregulation of GKRP and GK have yet to be resolved but may involve the protection of both proteins from degradation in their combined state.
In the heart, the IEVs for glycogen phosphorylase shifted to more acid positions consistent with phosphorylation and activation of the enzyme in response to low insulin signaling and potentially modifications of cellular Ca2+ handling in diabetes (42, 51). These results support the contention that the Ca2+ signaling apparatus may be dysfunctional in the diabetic heart.
In both heart and skeletal muscle, enolase was downregulated in the diabetic rat and the enolase IEV moved to a more acidic range consistent with phosphorylation. This phenomenon has not been previously reported in the literature and suggests that enolase is undergoing a posttranslation modification that could suppress this highly active enzyme. There is evidence that enolase expression is downregulated in a genetic diabetic mouse model (58) and in a dilated cardiomyopathy model (21). Thus altered expression and posttranslational modification of enolase may play a significant role in muscle energetics and diabetic adaptation.
In the pseudostarved state of diabetes the liver is producing large amounts of glucose and urea from lactate, glutamine, and alanine generated by catabolism in other tissues. In diabetes, gluconeogenesis can be increased as much as 12-fold (71). Gluconeogenesis from lactate requires 6 ATP per glucose generated, while glucose generation from alanine also requires 6 ATP per glucose generated plus 4 more ATP to generate urea from the residual nitrogen (11, 34). With regard to urea synthesis, a 70% increase in urea synthetic rate has been reported in a rat model of diabetes (3, 60) as well as in human insulin-dependent diabetes mellitus (2) that correlates with the overall cachectic state. These observations suggest that urea and glucose synthesis are major driving forces for the increased capacity to generate ATP via oxidative phosphorylation and FAO in diabetes. This is graphically represented in Fig. 4.
With respect to gluconeogenesis there was a large increase in two key rate-limiting steps, phosphoenolpyruvate carboxykinase and pyruvate carboxylase, consistent with prior studies (4, 9), while the export of oxaloacetate from the mitochondria for glucose synthesis may be enhanced by the previously mentioned increase in cAT and mAT (Fig. 3). The remaining detected enzymes in gluconeogenesis catalyze near equilibrium, bidirectional reactions participating in both glycolysis and gluconeogenesis. However, diabetes increased glyceraldehyde-3-phosphate dehydrogenase, which is somewhat surprising for an enzyme that participates in both gluconeogenesis and glycolysis.
We also observed upregulation of the enzymes associated with scavenging of GABA for gluconeogenesis. The liver takes up plasma GABA (50) under control conditions with a liver-specific GABA transporter (8). The scavenging pathway for GABA to succinate via aminobutyrate transaminase (ABT) was enhanced in diabetes, where plasma GABA concentration is decreased (1, 22, 41). These data suggest that GABA may be a substrate for gluconeogenesis in diabetes.
Urea cycle.
There was a general upregulation of the enzymes responsible for the elaboration of urea, including carbamoyl phosphate synthase (CPS), ASS, and ASL, with the exception of arginase and ornithine carbamoyltransferase (OCT) (Fig. 4), consistent with many previous studies in a variety of models (7, 40). OCT can support increased flux without an increase in enzyme level, possibly because of the activation by Mn2+ (68). Arginase also demonstrates linear first-order kinetics to increasing concentrations of plasma arginine (65). Thus the present studies confirm a large body of work demonstrating upregulation of urea cycle enzymes in diabetes.
A striking isoelectric focusing shift in urocanase, catalyzing the second step of histidine degradation, in the liver, is of interest for future study. This may be related to the general upregulation of amino acid oxidation in the diabetic state. No posttranslational modifications of urocanase have been reported. Other upregulated enzymes associated with amino acid oxidation included glutamate/oxaloacetate transaminase, argininosuccinate synthase, 4-aminobutyrate aminotransferase, glycine methyltransferase, serine/threonine dehydratase, glutaminase, phenylalanine-4-hydroylase, glutaryl-CoA dehydratase, aminoadipate semialdehyde synthase, and alanine aminotransferase.
Reactive oxygen species.
Protein oxidation and oxidative damage have been previously associated with diabetes (16, 55). The enzymes responsible for scavenging ROS in the cytosol [catalase, PRX-6-1-4, Cu-superoxide dismutase (CuOD), and glutathione S-transferases] were downregulated or unchanged without any significant change in enzymes responsible for scavenging ROS in the mitochondrial matrix [PRX-5, Mn-SOD, and glutathione peroxidase (GPX)-1] (Fig. 4). These data suggest that the cytosol would be more susceptible to ROS damage in diabetes due to a downregulation of ROS scavenging pathways. The liver ROS production would likely be higher (44), because of a high concentration of reduced substrates and enhancement of electron transport elements. The suppression of cytosolic ROS scavenging pathways may again reflect the disconnected signaling of the pseudostarved state of the liver. In the normal starved state ROS production is reduced (10, 69), while ROS scavenging mechanisms are unaffected or decreased (23). On the basis of these observations, we speculate that in the altered signaling milieu of diabetes the cytosolic ROS scavenging enzymes are downregulated in anticipation of a normal starved state reduction of ROS production. In diabetes the liver is adjusting its ROS scavenging capacity as if in a starved state, while the ROS production rate is actually increasing. This would result in the marked increase in free radical protein damage associated with diabetes (16, 55). As such, ROS metabolism dysregulation could contribute to the excessive protein oxidation pathology associated with diabetes.
Methionine cycle.
The proteomic data revealed upregulation of many enzymes of the methionine cycle. This may relate to the body wasting characteristics of uncontrolled type 1 diabetes and a need for increased capacity for the catabolism of choline derived from the breakdown of phospholipids (Fig. 4). The net effect of the observed upregulation of enzymes of the methionine cycle would support this catabolism. These findings suggest that this could be an important adaptation in the cachectic diabetic state. The observed upregulation of choline oxidase and betaine hydroxymethyltransferase would increase the transfer of methyl groups of choline into the methionine cycle. Upregulation of choline oxidoreductase (CO) and betaine-homocysteine S-methyltransferase (BHMT) shunts methyl groups into the methionine cycle. Upregulation of glycine-N-methyltransferase (GNMT) advances the synthesis of sarcosine, which must be converted to glycine with formation of a one-carbon THF derivative. In the presence of adequate THF, the cycle would net one serine and either two formate or two formyl-CoA per choline metabolized. Since serine is a gluconeogenic amino acid and serine dehydratase was found to be upregulated, metabolism of choline produced by wasting of tissues of the body supports the synthesis of glucose by this pathway. If THF should happen to be limiting, then the result would be one serine plus two molecules of formaldehyde per choline. This is of interest since formaldehyde is elevated in serum and urine of diabetic patients (14) and is suggested to play a role in retinopathy and vasculopathy that accompany diabetes (70). The source of formaldehyde at present is believed to be increased activity of the polyol pathway in peripheral nerves generating high concentrations of fructose that nonenzymatically glycosylate proteins that are broken down to formaldehyde (6, 62). It is of note that neither aldose reductase inhibitors (29, 54) nor sorbitol dehydrogenase inhibitors (56) have been shown to ameliorate all of the complications attributed to this pathway, except in concentrations high enough to prevent all flux through the sorbitol pathway with aldose reductase inhibitors (46). In addition, in this data set there was decreased sorbitol dehydrogenase as well as aldose reductase (lower-confidence identification), and both were highly significant (P < 0.0053). These data demonstrate another potential source of the formaldehyde in diabetes that warrants evaluation, and suggest a possible metabolic adaptation that has been unrecognized in the literature. There are studies that have demonstrated upregulation of elements of the methionine cycle (52, 66), but these studies were unable to evaluate the entire pathway concurrently and that is a particular strength of the present data.
Protein differences not associated with canonicals.
The advantage of this study was evaluation of multiple proteins and associated metabolic pathways simultaneously in one diabetic setting. There were many previously unrecognized protein expression differences identified in this data set; specifically, there were 38 proteins that were significantly differentially regulated in the liver and 12 in the heart that have not previously been reported. Several of these proteins were discussed in the canonical pathway discussion above; however, some novel observations might warrant further investigation. As an example, the tripartite motif-containing protein 72 was altered in both heart and skeletal muscle with diabetes. This protein has no known function but contains zinc finger domains that may imply a transcription factor activity. 3-Oxo-5-β-steroid 4-dehydrogenase is necessary in bile acid synthesis (31) and as such may have a role in the hypercholesterolemia that accompanies diabetes. d-Dopachrome tautomerase was downregulated in diabetes and is an inflammatory mediator important in T cell and macrophage migration inhibition (47, 63) with no established role in diabetes beyond that ketones are effective inhibitors of this enzyme (19). Liver calcium/calmodulin-dependent 3′,5′-cyclic nucleotide phosphodiesterase 1C (CDCDP) was significantly downregulated in diabetes, suggesting a modification of the calcium signaling/handling pathways in diabetic liver as described for the heart.
These data give an overall impression of the enzyme perturbations and global function of the liver during the diabetic state. An overview of metabolism of the diabetic liver is graphically depicted in Fig. 5, in an attempt to generate a more systems-oriented examination of the consequences of diabetes in the liver. The overall increase in ATP production capacity is used for glucose and urea synthesis utilizing the incomplete oxidation of fatty acids to ketone bodies and short-chain fatty acids likely due to a downregulation of CS. The increase in ROS generation associated with oxidative capacity is not balanced by an increase in ROS scavenging pathways. This may lead to an increase in ROS and protein oxidative damage. The processing of choline from phospholipids is enhanced and may fuel the generation of formaldehyde through the methionine cycle. Like most discovery approaches, more hypotheses were generated by these data than solutions to problems; however, this screening information is critical for the validation of existing models and provides novel insights for the formulation of new hypotheses on the biochemical adaptations associated with diabetes.
↵1 The online version of this manuscript contains supplemental material.
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- Copyright © 2009 the American Physiological Society


text:
Submitted: June 28, 2010 - 10:06pm
Originally published: June 28, 2010
Last updated: June 28, 2010 - 10:08pm
Originally published: June 28, 2010
Last updated: June 28, 2010 - 10:08pm
Source: National Public Radio
Author: Heather Chaplin
Quest to Learn, 225 East 23rd Street, New York, NY, 10010, United States
A public school in New York City has taken the video game as its model for how to teach. Students use video games and design them as part of their classes. As Quest to Learn is wrapping up its first year, those behind the program say game-based learning is integral to 21st century literacy.
Links to Sources
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- FTC Helps Prepare Kids for a World Where Advertising is Everywhere
- Six technologies soon to affect education
- US Department of Education Announces $25 Million for Science and Literacy-Themed Television and Digital Media
- Consumer Demand for Digital Learning Games, Simulations Growing Worldwide
- Using Video Games as Bait to Hook Readers
- UK Pupils to study Twitter and blogs in primary schools shake-up
- iPad Age Prompts India to Sell $22 Student Tablet to Boost Usage
- Microsoft Lobbyist at White House Event
- Connect to Compete
- Coding Class, Then Naptime: Computer Science For The Kindergarten Set
- Education and the National Broadband Plan
- CPB and PBS Partner with 11 Communities as Demonstration Sites for Ready To Learn Media
- Children's Agenda for Digital Opportunity
- Public Media Closes the Early Literacy Gap
- Why more schools aren't teaching web literacy -- and how they can start

subset: commoncrawl


text: Dear GHC-ers, I'm working on building an interactive environment around the composition of expressions. Users can type in (i.e. give strings of) expressions and can then use these expressions to produce other expressions. I'm close to having a working GHC-API binding for this. The resulting types, however, still contain some things I don't quite understand. Any help would be appreciated. Below, I've included the function exprFromString which should parse, rename and typecheck strings to Id-things and give their type (although, ideally, the idType of said Id-thing should be the same as the type returned). This function lives in the IA (InterActive) monad; a monad that is a GhcMonad and can lift monadic computations in TcM into itself using liftTcM (which uses the initTcPrintErrors and setInteractiveContext functions similarly to TcRnDriver.tcRnExpr). Near the end of the function, debugging output is produced. This output confuses me slightly. Here is the output for the three inputs "map (+1) [1..10]", "5" and "\\x -> x": map (+1) [1..10] pre-zonk: forall b. (GHC.Enum.Enum b_i, GHC.Num.Num b_i) => [b_i] post-zonk: forall b. (GHC.Enum.Enum b_i, GHC.Num.Num b_i) => [b_i] idType: [b_c] tidied: forall b. (GHC.Enum.Enum b_i, GHC.Num.Num b_i) => [b_i] 5 pre-zonk: forall a. GHC.Num.Num a_d => t_c post-zonk: forall a. GHC.Num.Num a_d => t_c idType: a_b tidied: forall a. GHC.Num.Num a_d => t_c \x -> x pre-zonk: forall t. t_e post-zonk: forall t. t_e idType: forall t. t -> t tidied: forall t. t_e The zonking and tidying part of the type-checking process are still a bit unclear to me and I suspect the problems arise there. It looks to me that the type variables in the quantifications are different ones from those in the pi/rho-types. I had expected the types to only contain the variables over which they are quantified, so e.g. in the map-example, I had expected "forall b . (GHC.Enum.Enum b, GHC.Num.Num b) => [b]" Can anyone explain what I'm missing? Regards, Philip exprFromString :: String -> IA (Id,Type) exprFromString str = do dfs <- getDynFlags let pp = showSDoc dfs . ppr pst <- mkPState dfs buf <$> newRealSrcLoc {- Parse -} (loc,rdr_expr) <- case unP parseStmt pst of PFailed span err -> throwOneError (mkPlainErrMsg dfs span err) POk pst' (Just (L l (ExprStmt rdr_expr _ _ _))) -> do logWarningsReportErrors (getMessages pst') return (l,rdr_expr) POk pst' thing -> throw $ maybe EmptyParse (const NonExpressionParse) thing liftTcM $ do fresh_it <- freshName loc str {- Rename -} (rn_expr, fvs) <- checkNoErrs $ rnLExpr rdr_expr {- Typecheck -} let binds = mkBinds fresh_it rn_expr fvs (((_bnds,((_tc_expr,res_ty),id)),untch),lie) <- captureConstraints . captureUntouchables $ tcLocalBinds binds ((,) <$> tcInferRho rn_expr <*> tcLookupId fresh_it) ((qtvs, dicts, _bool, _evbinds), lie_top) <- captureConstraints $ simplifyInfer True False [(fresh_it, res_ty)] (untch,lie) let all_expr_ty = mkForAllTys qtvs (mkPiTypes dicts res_ty) say str say $ " pre-zonk: " ++ pp all_expr_ty zonkTcType all_expr_ty say $ " post-zonk: " ++ pp all_expr_ty say $ " idType: " ++ pp (idType id) say $ " tidied: " ++ pp (tidyTopType all_expr_ty) return (id,all_expr_ty) where say = liftIO . putStrLn buf = stringToStringBuffer str freshName loc str = (\u -> mkInternalName u name loc) <$> newUnique where name = mkOccNameFS varName $ fsLit $ "it" ++ show (lineOf loc) isVarChar c = isAlphaNum c || c == '_' || c == '\'' lineOf (RealSrcSpan s) = srcSpanStartLine s lineOf _ = -1 mkBinds :: Name -> LHsExpr Name -> FreeVars -> HsLocalBinds Name mkBinds nm e@(L l _) fvs = HsValBinds $ ValBindsOut [(NonRecursive, unitBag the_bind)] [] where the_bind = L l (mkTopFunBind (L l nm) [mkMatch [] e emptyLocalBinds]) { bind_fvs = fvs }


text:
* Ramona calling Scott "Another Evil Ex waiting to happen."
** She manages to drive the point home even harder with four simple words: "Here's your damn list."
* The part where Scott breaks up with Knives and then the song Teenage Dream (by T-Rex, NOT Katy Perry) starts playing as he's on the bus was also really sad too.
* Also, the part where Scott sees Knives in the crowd after defeating the Twins.
** Beautifully accompanied by Broken Social Scene's lilting, wistful ''Anthems for a Seventeen Year Old Girl.''
* The whole part after the final fight up to the credits. [[spoiler: Knives and Scott seems to finally kick in and it looks like they're gonna try again (Knives even commenting that his hair has grown longer, and cue Scott with his hat running gag. She just dismisses it and tells him he should cut it, to which he agrees). Ramona turns around and starts walking away, and Scott and her says their "final" goodbyes. Knives, however, doesn't want Scott to suffer and stay that way, so she tells him to get her back. Cue Scott and Ramona, walking towards a subspace door, ending the scene in a sunset. The REAL Tear Jerker, however? Scott's Ramona cute song, in a beautiful and sad reprise, until the credits, and the "CONTINUE?", show ups.]]
** The countdown just before the credits can make one's heart ache for some reason.
* Knives's little "Oh" after [[spoiler: Scott breaks up with her.]] She slowly looks down at her own heart right after that - you can almost ''see'' it appearing in cartoon form and breaking in two, even if the movie itself doesn't go ''that'' far.
* ''Ramo-o-ona, oh, Ramona...''


text:
Federer says he can dominate men's tennis again
The tennis player who lost his No 1 ranking last year says he is feeling much better about his health and feels more confident.DOHA – Former No 1 Roger Federer believes he can dominate men's tennis again.
The 27-year-old Swiss star, who is one short of Pete Sampras' record of 14 Grand Slam titles, said Monday he is looking forward to a big year in 2009.
"This could be a very meaningful season for me," Federer said. "I can equal Pete Sampras' record. I can add another Wimbledon title and this is something that is very dear to me. I can also win my first French Open title, so I am looking forward to my season."
Federer was slowed by mononucleosis last year and lost his No.1 ranking to Rafael Nadal.
"I feel much better this year," Federer said. "Last year, I couldn't practice the way I wanted to because of health reasons. But this year I have trained really well and I feel very confident about myself. I have less pressure now and it is not a bad thing."
Federer plays Potito Starace of Italy in the first round at the season-opening Qatar Open on Tuesday as he builds toward the Australian Open, which starts on 19 January.
"Obviously, I can dominate the game," he said. "I have worked hard at my game and when you do that, things become easy for you. If I can play two good matches then I will feel really good going into the Australian Open."
The second-ranked Federer was eclipsed last year by Nadal, who won Wimbledon, the French Open and the Beijing Olympics gold medal.
"Rafa is my main rival," Federer said. "We have been doing it for so many years and have had great matches."
Andy Murray beat Federer and Nadal last week at an exhibition tournament in the United Arab Emirates. Federer could face Murray in the semi-finals in Qatar after having defeated the Briton in the US Open final for his only Grand Slam title of 2008.
[AP / Expatica]


text:
As with the Frank Wills Team the week before, the Cornwall Bowl team were defeated at the semi-final stage. The match against Mullion was played at Trevose in sunny, dry conditions, but with a very strong wind.
The match was played in fine spirit, with Mullion running out winners by the score of 4 matches to 1.
Playing conditions were most definitely challenging, and the Trethorne players would be the first to admit the match was won and lost on and around the greens. A special mention is reserved for Ian Burn and Miles May, who won their first match of the campaign, 2&1.


text:
Get an overview of this hotelQuiet oceanfront resort in middle of Miami Beach
Popular property highlights
Water all around
Located in a less populated stretch of mid-Miami Beach, this resort faces the Atlantic Ocean on one side and the Intracoastal Waterway on the other; glitzy South Beach is 3 miles away.
Oceanside pool, dock
A large pool with rental cabanas overlooks the beach; guests can rent boats and watersports equipment at a private dock. The Miami Beach Resort also offers spa services and a health club.
Rooms with views
Floor-to-ceiling windows look out to full or partial water views. Guestrooms also offer pillowtop mattresses, wireless Internet access, and multi-line phones.
Guest Highlights
- Good location
- Proximity to shopping
- Beach
- Pool
- Helpful staff
- Bathrooms could've been nicer
Miami Beach Resort
Hotel Amenities
Hotel Amenities
Miami Beach Resort is located on a quiet stretch of beach, three miles from South Beach attractions. Situated between the Atlantic Ocean and the Intracoastal Waterway, the resort features a private dock with boat and other water-sports rentals, and a large oceanfront pool with rental cabanas and concierge service. Airport transportation is available for a surcharge.
Guests can enjoy hair care and spa services (surcharges); a health club; indoor and outdoor dining options; complimentary children's activities; and golf privileges at Miami Beach Golf Club.
The resort's self-service business center offers printing, faxing, and computers with high-speed Internet access. Complimentary wireless Internet access is available in public areas.
Valet parking (surcharge) is also available.
- 24-hour front desk
- ATM/banking
- Year Built 1963
- Number of floors - 17
- Valet parking (surcharge)
- Bar/lounge
- Free newspapers in lobby
- Concierge services
- Coffee shop or café
- Conference space
- Dry cleaning/laundry service
- Elevator/lift
- Full-service spa
- Gift shops or newsstand
- Health club
- Free WiFi
- Free wired Internet
- Spa treatment room(s)
- Multilingual staff
- Total number of rooms - 424
- Poolside bar
- Porter/bellhop
- Safe-deposit box at front desk
- Number of restaurants - 3
- 24-hour business center
- Beach umbrellas
- Beach cabanas (surcharge)
- Beach sun loungers
- Smoke-free property
- Outdoor pool
- Wedding services
- Breakfast available (surcharge) hot and cold buffet
- Hair salon
- Pool umbrellas
- Number of meeting rooms - 6
Family Friendly Amenities
- Family Hotel
- Outdoor pool
- Free Wi-Fi
- Cribs/infant beds (complimentary)
Available in all rooms: Free WiFi
Available in some public areas: Free WiFi , Free wired Internet
Valet parking (surcharge)
Room Amenities
- Free weekday newspaper
- Air conditioning
- Phone
- Free toiletries
- In-room safe
- Desk
- Blackout drapes/curtains
- Cable TV service
- Television
- Pillowtop mattress
- In-room climate control (air conditioning)
- Coffee/tea maker
- Designer toiletries
- Free local calls
- Hair dryer
- Daily housekeeping
- Iron/ironing board
- Free WiFi
- Pay movies
- Premium TV channels
- Shower/tub combination
- Private bathroom
- Free cribs/infant beds
- Connecting/adjoining rooms available
- Rollaway/extra beds (surcharge)
Where to Eat
Hibiscus Court - Overlooks pool and garden area. Gourmet buffet breakfast served daily.
Seabreeze Bar & Grill - Located poolside. Covered or outside seating. Burgers, salads, sandwiches, and light fare. Daily lunch served.
Ocean Bar & Bistro - Fine-dining restaurant. American and island-themed cuisine. Nightly dinner served.
Poquito's To Go - Coffee bar with take-out breakfast items. Daily breakfast served.
Nearby Things to Do
The Miami Beach Resort is located between a white-sand Atlantic Ocean beach and the Intracoastal Waterway. Rental equipment at a private dock includes catamarans, personal watercraft, and scuba and snorkeling equipment. In addition, fishing charters and beach volleyball can be arranged (surcharges).
The large oceanfront pool provides rental cabanas. Pool concierge service includes magazines, newspapers, floating devices, games, snacks, towels, and lounge chairs. Children's activities are complimentary.
Guests have complimentary access to a health club with cardio equipment and free weights, lockers, and saunas.
Offering 18 holes of championship golf, Miami Beach Golf Club, three miles away, extends privileges to resort guests (surcharge).
The recreational activities listed below are available either on site or near the hotel; fees may apply.
- Golfing nearby
- Snorkeling nearby
- Kayaking nearby
- Sailing nearby
- Scuba diving nearby
- Water skiing nearby
- Bicycle rentals nearby
- Parasailing nearby
- Volleyball on site
- Boat tours nearby
Spa Services
- Sauna in spa
- Spa tub in spa
- Massages
- Body treatments
- Number of treatment rooms 4
- Facial treatments
If you have requests for specific accessibility needs, please note them at check-out when you book your room.
- Accessible bathroom
- Roll-in shower
Hotel Policies
Minimum check-in age is 21
Check-in time ends at midnight
Check-in time starts at 4 PM
Check-out time is 11 AM
Payment types
Children and extra beds
- Children are welcome.
- Kids stay free! Children 17 years old and younger stay free when using existing bedding.
- Rollaway/extra beds are available for USD 20.00 per day.
- Free cribs (infant beds)!
- Pets not allowed
You need to know
Extra-person charges may apply and vary depending on property policy.
- Deposit: USD 50.00 per night
- Resort fee
- Internet access
Optional extras
- Breakfast fee: USD 19.00++ per person (approximately)
- Valet parking fee: USD 32 per night (in/out privileges)
- Rollaway bed fee: USD 20.00 per day
Hotel Name
- Miami Beach Resort
- Resort Miami Beach
- Miami Beach Resort Spa
- Miami Beach Hotel Miami Beach
We should mention
A resort fee is included in the total price displayed. Children 17 years old and younger stay free when occupying the parent or guardian's room, using existing bedding. Only registered guests are allowed in the guestrooms. The property has connecting/adjoining rooms, which are subject to availability and can be requested by contacting the property using the number on the booking confirmation.


text:
I'm considering chin lipo, I am only 33 so i do not have sagging issues, but if I lose weight later on, will the shape of my face get weird?
What if I Lose Weight After Chin Lipo?
Doctor Answers 13
Revisional Surgery May Be Necessary When Loose, Saggy Skin is a Result of Weight Loss
It's not unusual for patients to lose weight following neck liposuction. When weight loss is minimal to moderate, the surgical results of neck liposuction are rarely affected. When patients experience massive weight loss following neck liposuction, dramatic changes can occur in the neck region. These may include a hollowing of the central neck and the development of loose saggy skin.
It's important to understand that no two patients are alike and for this reason, treatment needs to be individualized. In the vast majority of cases we prefer that patients are within 10-15% of their ideal body weight.
Unfortunately, this isn't always possible and some patients proceed with neck liposuction before they've reached this goal. Under these circumstances, subsequent weight loss can cause secondary problems which may require revisional surgery.
It's safe to say that the majority of neck liposuction patients aren't affected by normal fluctuations in weight. When patients undergo massive weight loss, all bets are off and secondary surgery may be occasionally necessary. Under these circumstances, consultation with a board-certified plastic surgeon is appropriate.
Best to lose weight before, not after Chin or neck liposuction
Generally speaking, significant or major weight loss results in laxity of the skin or sagging of the skin as the fat beneath the skin is lost.
Thus if a patient is overweight, Plastic Surgeons usually recommend weight loss to near normal weight before doing liposuction.
If you are not significantly overweight now, then 5 or 10 pounds of weight loss will not likely cause much of a change in skin tone after liposuction.
However it is best to be able to keep your weight stable after liposuction.
Newer Laser Assisted Liposuction techniques such as Slim Lipo have the added benefit of producing skin tightening as well as fat removal. These techniques are particularly good for the chin and neck area (see pics in reference below.)
This additional skin tightening during the course of chin liposuction might offer some added protection against skin laxity if you lose weight after chin or sub mental neck liposuction.
Learn more: http://drseckel.com/non-surgical-procedures/laser-double-chin-removal-the-nonsurgcal-neck-lift/
Don't worry
Liposuction of the chin or submental area is a very gratifying procedure with sometimes dramatic results when done on the right patient. As long as the skin elasticity of the neck is good then the skin will re drape and shrink a bit after the procedure to take up the slack if you will. The goal of liposuction in the submental area is to pretty much remove all the fat and make this area flat. If you lose or gain weight later on in life there should not be a significant change in the area suctioned because the majority of the fat cells have been removed.
Good luck with your surgery.
You might also like...
Some changes in chin lipo results after extreme weight loss
Extreme weight loss might affect your facial features but not a 5-10 pounds weight loss. The main areas are the cheeks and the neck if you have excess fat there.If you already had lipo of the neck no major difference should happen.
Learn more: http://newportplastic.com/face-lift/
Losing weight after chin lipo
Lost Weight after Chin Lipo
Prior to liposuction you should be at a stable weight. 5-10 lbs lost after your procedure wont affect your face much, however extreme weight lose will affect your features or possibly give you a "saggy" appearance after surgery as the fat was already removed leaving a pocket. Before getting this procedure, get to a desired weight and maintain it for 3-6 months.
Learn more: http://www.capefearaesthetics.com/details/plastic+surgery-19/
It might....
Lipo is best when you are at a weight you can maintain more or less over time. It is best to get to that "stable weight" point before surgery.
Best Regards,
John Di Saia MD
Losing weight should not affect your neck appearance.
Losing weight should not affect your neck appearance much at all. Some families tend to "carry" the weight in certain areas; the neck is one of those areas.
Make sure you see someone that offers all different treatments including liposuction, SmartLipo, and neck lifts so that you can get an honest opinion about which procedure you need.
Weight loss after chin liposuction
If you are significantly overweight and you plan on losing a lot of weight, I recommend you hold off on liposuction to you chin or anywhere else until you have maintained your lower weight for 3 - 6 months. I think even 10 or 15 pounds can profoundly influence the fat under the chin in some individuals. You would not want to be in the situation of having liposuction, losing weight and then having the chin and neck area looking gaunt.
Lisa Lynn Sowder, M.D.
Weight loss after Chin liposuction
After Chin liposuction, if you continue to lose weight you should not affect your ultimate appearance. The two most important factors for a successful result after liposuction to this area are the quality of your skin in the neck and your compliance with the postoperative care. Your best bet is to be evaluated by a board-certified plastic surgeon who has a great deal of experience with chin surgery and liposuction. They will be able to determine the quality of your skin and if your skin will contract sufficiently to create a great result after the fat has been removed.
These answers are for educational purposes and should not be relied upon as a substitute for medical advice you may receive from your physician. If you have a medical emergency, please call 911. These answers do not constitute or initiate a patient/doctor relationship.


text:
Beyoncé Knowles 'planning wig suitcases'
Beyoncé Knowles has allegedly invested in three different suitcases of hair supplies for her upcoming world tour.
The international superstar's hotly-anticipated shows will kick off in Belgrade, Serbia on April 15 before winding up on August 3 in Brooklyn, New York.
Beyoncé is keen to impress fans with her look and has allegedly invested in three different suitcases of hair supplies.
"Bey's stylist has one suitcase of weaves, one of lace fronts (a wig that attaches to the front of the head) and one of clip-ons," a source revealed to British magazine Heat.
"They are colour co-ordinated and vary in volume. When she's feeling fierce, she'll go for one that's bright yellow. When she's feeling more demure, she's got her dark brown poker-straight hair like [US First Lady] Michelle Obama's original style."
It was recently revealed that Beyoncé has asked British designer Julien Macdonald to design her costumes for the 59-date tour.
"Julien is a huge Beyoncé fan so he's really excited about producing some custom-made, bespoke outfits. This is a huge coup for him. Bey loves his work and has often worn his outfits," a source told the Daily Mirror last week.
The fashion designer - who has collaborated with the char-topping star on a number of outfits - has always gushed about Beyoncé's style.
"There's nothing better than seeing somebody you admire, or someone who embodies Julien Macdonald," he said. "I like girls like Beyoncé or Kristen Stewart, they are amazing women. Iconic. They are part of a new generation. When she started wearing my clothes it was amazing!"
© Cover Media


text:
10 Weeks Later: The East Harlem News is Born!
December 20, 2012
This semester, the Washington Square News partnered with the nonprofit organization Citizen Schools to build and teach a journalism curriculum to a class of sixth graders at Isaac Newton Middle School in Harlem.
After 10 weeks of hard work, the class finally finished their project: a newspaper for the East Harlem Community.


text:
50 Years of Truely Sustainable Agriculture to be Celebrated Next Year
"No-till" Soybeans Following Wheat
There is a sub-set of farmers who have been practicing a much more sustainable form of agriculture for decades and we are coming up on the 50th anniversary of it's beginnings. I want to start writing about this event early because many environmentally-conscious folk are not aware of this hugely significant "revolution" that has occurred in agriculture over the past half century. No, I'm not talking about "Organic Farming." That movement is about a decade older in the US and has had a much smaller impact. I'm talking about "No-till" agriculture and variants that are focused on reducing the amount of "tillage" or "plowing" that are needed to farm.
For many centuries, plowing and tillage have been key practices for agriculture.
That was how a good "seedbed" was prepared and that was how weeds were controlled. But just because a practice is ancient does not mean it is good. Plowed land is much more subject to water and wind erosion and plowing combined with annual cropping has lead to the depletion of carbon stores in our soils.
History of No-till
In 1943, Edward Faulkner wrote a book titled "The Plowman's Follyin which he questioned the reasons for plowing. That was a very controversial idea at the time, but by 1960, researchers began to try to find a way to farm that didn't involve disrupting the soil. The crop "residue" was chopped and left on the soil surface. The next crop was "drilled" into that matrix. Weeds were not controlled by mechanical disruption of the soil, but by the use of herbicides (and yes, there are safe and environmentally benign ways to do that). Between the work of USDA and University scientists, progressive growers and equipment and chemical companies, this concept was turned into an economically viable option that has now been adopted on around 66 million acres of US cropland (20% of the total) and on 260 million acres world-wide (excellent but fairly large slide set about this from no-till experts Rolf Derpsch and Theodor Friedrich). This system has proven to have the following advantages:
- Water and wind erosion diminished to near zero (with the added benefit of reduced fertilizer and pesticide residue movement that was associated with that soil movement)
- Reduced fuel use and the greenhouse gas emissions associated with it (e.g. 66% reduction in corn)
- Labor efficiency (e.g. an individual grower can plant more acres or can reduce labor expense)
- Improvements in soil quality in terms of aggregate stability, rainfall capture efficiency (less runoff) and water holding capacity
- Increased soil carbon (particularly if paired with "cover-cropping") that can "sequester" carbon dioxide from the atmosphere
- Increased biodiversity both above and below ground (e.g. earthworms)
- Better plant association with mycorrhizae (soil fungi which help plants obtain nutrients)
- Lower nitrous oxide emissions on most soils (a potent greenhouse gas)
No-till and other "Conservation tillage" systems combined with cover-cropping is the most sustainable option for agriculture today. Organic producers (who are more likely to practice cover cropping, to their credit) don't really have the option of reduced tillage. There are some efforts in that direction, but they are not practical on any significant scale. Organic has been focused on building soil organic carbon and that is an excellent thing to do, but part of the way they do it, importing large quantities of compost or manure, is problematic from a greenhouse gas perspective as I described in a previous post. Reduced tillage and cover cropping is much more like the way that soils are built in a natural system.
So in the spring of 2010 those of us who are focused on sustainable agriculture will celebrate the work that was started a half century ago. Maybe a new market of carbon off-sets will help spur even more conversion.
No-till Soybeans Image from NRCS. Traditional plowing image from IRRI.
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text:
Have Ford Questions? Ask a Mechanic for Answers ASAP
Welcome to Justanswer
The problem can be from being low on fluid the first thing to do is check the level
i did check it it seems ok to me
i parked on a incline and when i went to put it in drive i was kind of harder because ofit being on a hill theni got it in frive i stepped on gas and it woldnt go forward or backwarf
i hear a noise
Does it feel like the brake is on or does it feel like its still in park
Does it feel like a brake is on or does it feel like its in park still
feels like it is still in park
can it be my cable wire or my sitch
Sounds like you broke park pin on the transmission cable
Put it in drive or neutral does it move
no it doesnt do anything
it might coast a little bit but thats it
So it does move?
not by the engine but by hand it will roll
is it a big job
is it a big job to do
You need to look and see if anything moves when you try to shift it under the car the linkage is probably broke or came apart
This is not that bad and can be fixed easily
one more question my engine light is on and when they put it to machine it comes up lean wht can that be
they fixed the gas valve and they out new plugs and wires on
This can be a few things a start of a bad catalytic convertor A bad O2 sensor and can be caused by the Mass Airflow Sensor being bad
Those are the most common causes
ok so u dont think i would need a new transmittsion
one more thiing my brake light and abs light comes on once in a while
this van drives me crazy
that can be a cause of a loose connection on one of the wheel sensors I would inspect them and clean as needed
ok thank you so very much i was having a heart attack lol
No problem please hit accept so I get paid for my time and effort. Thanks have a great night and HAPPY 4th of JULY Danny
Let me know if you need anything else if not please hit accept.. Thanks Danny


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Thoughts to help your child after a tragedy
In light of Friday's school shooting in Connecticut, we asked a school counselor, Jefferson Elementary's Melissa Rather Brown, to give us some ideas for parents having difficulty talking to your children about tragedies. She compiled the following tips from various sources.
Limit media exposure. Adults often keep looking for answers and new developments in the media and may not realize how much their children are absorbing. For now, keep the TV and computer off. Remember that the computer home page that pops up often has the latest news and children often have exposure to this without adult supervision. Seeing repeat images and hearing the news over and again often makes children think it is happening again instead of a repeat of the same story.
Children will naturally feel insecure and even fearful. The buildings and people shown may look familiar and children may think this is a place or a face they recognize. Don't negate these feelings, but reassure them that these instances are SO RARE and that you and their school personnel do everything they can every day to keep them safe. Trying to rationalize statistics and probability with children will have little or no impact and not reassure them. It is not dishonest to say you don't imagine it will happen to them. It is so much more important to express your hope as a certainty in order to give children a feeling of stability and safety.
If children mention this tragedy or ask questions, ask them how they feel about it and answer the questions they ask. Let them talk, but don't force them to talk if they are not ready or don't have a grasp of the events. Be honest but measured; don't stir anxiety by bringing up feelings that children have not expressed having.
For older children, be present and available, but again, don't force them to talk if they are not ready. It is not indifference, but is more often their developmental level that does not create an immediate connection to something that is both geographically distant and not a part of their normal experiences. Try to reengage children in normal everyday activities as much as possible. Be available to answer questions as they come up, reassuring them that you and their schools are possibly even more aware and taking more steps to ensure their safety.
If a child at any age continues to express anxiety or fear over a period of time beyond the media flurry of horrific stories, please consider talking to your pediatrician about seeking additional help. Most children will move past the immediate insecurity and not dwell on it unless they are directly in the affected community. Watch for signs that might indicate your child needs additional time and help to deal with these tragic events. Above all, reassure your children that you are there to love them and take care of them.


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Health Care Reform
As part of efforts to address the health insurance needs of members, NAR has advocated for reform of the health insurance markets that provide coverage to the self-employed and small employers. Here you'll find information about new health care reform laws that will make a major change to many aspects of our health care system.
More than one-quarter of REALTORS® are uninsured. As part of efforts to address the health insurance needs of members, NAR has advocated for reform of the health insurance markets that provide coverage to the self-employed and small employers for more than nine years.
Among the legislative approaches that NAR has advocated are small business health options plans (SHOP), small business health plans (SBHPs) and association health plans (AHPs). NAR continued to represent the interests of the REALTOR® community in the recently completed comprehensive health reform debate.
Patient Protection and the Affordable Care Act
The Affordable Care Act (ACA) (link is external) was signed into law on March 23, 2010. On June 28, 2012, in a 5-4 decision, the U.S. Supreme Court upheld the constitutionality of the ACA.
The Act is complex and multifaceted; of particular importance to REALTORS® and those who must purchase their own health insurance are the ACA underwriting and rating reforms that help to conform the characteristics of health insurance plans purchased by individuals with plans that are traditionally available from employers. Many provisions of the law were effective immediately. The last provisions of the ACA were scheduled to take effect on Jan. 1, 2015, though that timeline was extended for some .
Legislation, Rulings & Timeline
NAR has put together information on the two bills that comprised the health reform legislation, including Congressional roll call votes, summaries of the bills' provisions, and the text of the Supreme Court's ruling on the Affordable Care Act.
Want to learn more?
Both NAR and independent organizations have created a wide array of materials to help explain the provisions of the new health law, as well as provide more detailed commentary. Some of the most helpful materials can be found at:
- Healthcare.gov (link is external)
The federal government website that provides health insurance choices for individuals, families and small businesses.
- Kaiser Health News FAQs (link is external)
Kaiser Health News has put together a list of hot topics on the health law, insurance, medicare and medicaid, and other FAQs.
- Kaiser Family Foundation FAQs (link is external)
The Henry J. Kaiser Family Foundation compiled an extensive list of frequently asked questions to help you understand the eligibility, requirements, and responsibilities under the Affordable Care Act.
- IRS Individual Mandate FAQs (link is external)
Starting January 2014, you and your family must either have health care coverage, have an exemption from coverage, or make a payment when you file your 2014 tax return in 2015.
- IRS Employer Mandate
The Obama Administration has delayed the implementation of the employer mandate until January, 2015 to give businesses additional time to comply with the Affordable Care Act (ACA). Review the IRS tax provisions for employers (link is external).
- NAR FAQ: The 3.8% Tax
A 3.8 percent tax on investment income was created as part of the Affordable Care Act (ACA) (link is external). This tax WILL NOT be imposed on all real estate transactions, a common misconception. Please view our frequently asked questions to help you understand this tax provision.
- State Actions to Address Health Insurance Exchanges (link is external)
The National Conference of State Legislatures has a map showing the decisions made by each state regarding the the state Health Exchange marketplaces that serve as the new insurance marketplace for individual and small business insurance coverage.
- How to Apply for An Exemption (link is external)
The Centers for Medicare & Medicaid Services published a fact sheet released by the IRS on the various exemptions from the individual mandate, such as being unable to find affordable health insurance or living in a state that did not expand Medicaid.
Additional Resources
HealthCare.gov - Access to Federal and State Health Insurance Exchanges
Employers Responsibilities under the Affordable Care Act (IRS)
Affordable Care Act Tax Provisions (IRS)
REALTORS® Insurance Marketplace
How to Apply for An Exemption (Centers for Medicare & Medicaid Services)


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Mother stands behind preschool after two workers arrested
Sat, 29 Sep 2012 23:59:30 GMT —
Lori Turner is keeping her children in the Teaching Learning Center preschool in Myrtle Beach following the arrests of the director and an employee.
On Thursday, Mary Lynn Boggs was arrested for child negligence or abuse.
The arrest came after allegations surfaced that Boggs was covering children with blankets and holding them down for nap time.
Saturday, Brenda Thomas turned herself in for failing to report the negligence or abuse in Boggs' case.
According to state law 63-7-310, Thomas is required to report incidents of abuse to authorities.
Turner is the mother of two 5-year-old twins that have been attending the Teaching Learning Center at Ocean View Baptist Church since they were just weeks old.
Turner says she trusts the preschool completely with her kids.
"It's a learning environment, it's not a daycare," says Turner. "Kids don't sit there and play and watch TV. It's all very structured."
Turner says she will continue to keep her children enrolled despite the charges.
Ocean View Baptist Church did not return our phone calls Saturday.
Earlier this week, Pastor Everett B. Douglas released the following statement:
"Our primary concern is and always has been the safety and health of the children in our care at Ocean View Teaching Learning Center. The necessary agencies and individuals have been notified and this matter is under investigation. The teacher is no longer employed with Ocean View Teaching Learning Center. It is not appropriate to comment further with this matter under investigation."


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The Knee

The knee joint, medically known as the tibiofemoral joint, is the largest joint in the body. Two bones make up this joint; the femur and tibia. There is also a sesamoid bone, called your patella or kneecap, that resides above your knee joint. The knee joint is dependent on the muscles to move it and ligaments which surround it for strength and stability.

The Ligaments of the Knee:

Four ligaments are present in the knee joint: the medical collateral ligament, lateral collateral ligament, anterior cruciate ligament, and posterior cruciate ligament. These ligaments provide strength and stability to the knee joint.

The medial collateral ligament (MCL) is located at the inside of the knee joint. It extends from the medial femoral epicondyle to the tibia. This ligament prevents excessive abduction of the knee. It is a thick ligament that typically does not require surgery to repair if it becomes injured.

The lateral collateral ligament (LCL) is located at the outside of the knee joint. It extends from the lateral femoral epicondyle to the head of the fibula. This ligament prevents excessive adduction of the knee.

The anterior cruciate ligament (ACL) extends posterolaterally from the tibia and inserts on the lateral femoral condyle. This ligament prevents excessive anterior movement of the tibia under the femur.

The posterior cruciate ligament (PCL) extends anteromedially from the tibia posterior to the medial femoral condyle. This ligament prevents excessive anterior movement of the femur on the tibia.

Injury to Your Knee Ligaments

If you suffer trauma to your knee, you may injure one or more of your knee's ligaments. Signs and symptoms of a knee ligament sprain include:

Pain

Swelling

Loss of knee range of motion (ROM)

Decresed strength of the muscles surrounding your knee

If you suspect you have sprained a knee ligament, you must check in with your doctor right away to get a confirmed and accurate diagnosis and to get started on treatment right away.

Physical Therapy after Knee Ligament Sprain

Physical therapy is a good idea if you have sprained one of your knee ligaments. Your PT can work with you to restore normal strength and range of motion (ROM) to your knee and to help you return to maximal mobility and function.

Components of a rehab program after a knee ligament sprain may include:

Knee stretches

Knee strengthening exercises

Hip strengthening exercises

Balance exercises

Plyometric exercises

Knee bracing

Modalities to control pain and swelling that may occur

Your knee is a major weight bearing joint in the body and is responsible for walking, running, and rising from sitting. Injury to your knee can limit your ability to do these things. If you injure one or more of your knee's ligaments, you may benefit from physical therapy to help you regain normal mobility and function. That way you can quickly and safely get back to your normal activities.


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|No-Sew Gathered Curtains
Cut two panels of fabric -- each should be as long as your window and frame, plus 6 inches, and each one should be as wide as your window and frame.
At the top edge of each piece, fold over 4 inches towards the wrong side of the fabric. Iron the fold flat. If using fusible tape, follow the manufacturer's instructions and sandwich the fusible tape between the two pieces of the fold, and iron into place. If using fabric glue, follow the manufacturer's instructions and glue down the two pieces of the fold. You want to keep the fusible tape or glue near the cut edge of the folded over piece and leave at least 3 inches of the fold free as a casing to put the curtain rod into.
On the opposite end of each panel, fold over 1 inch of fabric -- make sure the folded over piece is folded towards the wrong side of the fabric. Use either the fusible tape or fabric glue to adhere the two pieces of the fold together. Now you have a bottom hem. Note: If the bottom edge of your curtains will hang behind furniture or otherwise not be seen, skip this step. Also skip this step if your curtains will be long enough to puddle on the floor -- just push the fabric edges underneath themselves and puff the fabric out a little bit for a lush, extravagant look.
Now slip the curtain rod into the top casings of each panel, and hang the curtains in the window. You can add ribbons or cording with tassels to tie back the curtains at the sides. If you want to block light, use heavy fabric or old blankets or just double layer the curtain fabric.
Gothic Martha Stewart Pages
starting : shopping : motifs : projects : weddings : notes


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With the regular season winding down, Iowa traveled to West Lafayette, Ind., on Saturday to take on Purdue with the Hawkeyes looking to rebound from a loss to Michigan State the previous week. Despite some sloppy play, Iowa (7-4, 4-3 Big Ten) was able to hold off a late Purdue (5-6, 3-4 Big Ten) rally attempt to walk away with a 31-21 victory. We will now take a look back and grade the Hawkeyes' performance.
After a rough outing in a home loss to Michigan State, James Vandenberg rebounded nicely against Purdue. He was more accurate with his passes, made few risky throws and overall looked more comfortable in the pocket than he did last week. Vandenberg was also finally able to get the monkey off his back by picking up his first road win as a starter at Iowa. His only major mistake was when he fumbled in his own end zone, which Purdue recovered for a touchdown to knot the score at 14-all. He also still tends to lock onto receivers at times instead of working through his reads, but it hasn't really cost him so far.
Overall, Vandenberg was 22 of 32 for 273 yards and three TDs. Those numbers could have been even higher were it not for several dropped passes by his receivers. Grade: B+
Running Back
It was another 100 yard outing for Marcus Coker as the sophomore methodically worked over the Purdue defense. He didn't have a lot of big runs (his longest was 14 yards), but he steadily pounded away at the Boilermakers with his usual physical style, picking up a large chunk of yards after contact. Coker carried the ball 30 times for 139 yards and a TD, his sixth straight game with at least one touchdown. It was also the seventh time this season he's gone over 100 yards and the third time he's carried the ball 30 or more times. Coker has now rushed for 1,297 yards in 2011, pushing him into sixth place on the Iowa single-season list.
Weeks of speculation as to whether or not Mika'il McCall would redshirt this season came to an end as the true freshman made his way onto the field in the second quarter. Unfortunately for McCall, it was not the kind of comeback he was hoping for. With Iowa sitting inside the Purdue 10-yard line, McCall fumbled on his second run of the day and did not receive another carry. Overall, he ran twice for 4 yards.
Although he didn't make a direct impact in the stat book, Brad Rogers once again did an excellent job blocking to help open things up for Coker and the run game. Grade: B+
Wide Receiver/Tight End
We might as well just call this the Marvin McNutt category from here on out. He did drop two passes, including a potential touchdown, but that doesn't wipe out what else he was able to do against the Boilermakers. McNutt hauled in nine passes for 151 yards and two TDs, including another highlight reel grab, and added to his growing number of school records, this time taking sole possession of the single-season touchdown record (12). This is in addition to his single-season record for receiving yards (1,240) and career records for touchdowns (28) and receiving yards (2,786).
McNutt is also poised to take possession of two more records as he sits just eight receptions shy of tying Kevin Kasper for single-season receptions (82 in 2000) and 11 short of tying Derrell Johnson-Koulianos for career receptions (173).
McNutt wasn't the only one making an impression against Purdue. C.J. Fiedorowicz continues to play a bigger role in the offense and caught three passes for 31 yards, including a 5-yard TD in the first quarter. Keenan Davis finished second on the team with seven receptions for 63 yards, while Kevonte Martin-Manley caught three for 28. As a whole, the receiving corps put up some solid numbers, but they still continue to drop far too many passes as they added at least four to an already too high season total. Grade: B
Offensive Line
The play of the offensive line was improved from last week's performance. While it certainly helps to not be facing Michigan State's massive defensive line, the Iowa O-line was able to generate a good push for much of the game and did a better job providing running lanes for Coker. Purdue was able to get to Coker for a loss of yards on several occasions, but overall the run blocking was good. Pass protection was also solid, and though Vandenberg was sacked three times, he usually had enough time to find his receivers. Grade: B+
Overall Offensive Grade: B Whether due to fumbles, dropped passes or some other miscue, Iowa let a lot of opportunities to score fall by the wayside. In the second quarter alone, Iowa had the ball at Purdue's 36, 9 and 49 and came away empty each time. That being said, the Hawkeye offense seemingly had little trouble moving the ball against the Boilermakers and was able to do enough to help push Iowa to victory.
Defensive Line
For a group that has been much-maligned this season, the defensive line turned in quite the performance on Saturday. Iowa was able to get into the backfield early and often, stopping Purdue from generating much of a running attack as the Boilermakers finished with just 101 yards on the ground. The D-line was also able to get through to Purdue quarterbacks Robert Marve and Caleb TerBush on several occasions, causing rushed throws when they weren't able to pick up a sack.
Mike Daniels and Broderick Binns were easily the stars of the defensive line on Saturday, turning in one of their best performances of the season. Daniels led the defense with a season-high eight tackles, including four for loss, and a career-high two sacks. Binns finished second on the team with seven tackles and a sack and was constantly in the Purdue backfield. Tom Nardo finished with four tackles (1 TFL) and broke up one pass, while Lebron Daniel had two stops and Steve Bigach had one. Grade: A
The linebacker corps played well against the Boilermakers, teaming with the defensive line to hold Purdue to just over 100 yards rushing. The tackling was better, they shed their blocks well for much of the game and they were effective in pass coverage as well.
With injuries that forced James Morris and Tyler Nielsen to the sidelines for stretches, Anthony Hitchens saw significant playing time and responded with a good performance, coming up with three tackles to go along with a fumble recovery. Morris led the group with four tackles, while Nielsen and Christian Kirksey finished with three stops with Kirksey adding a pass breakup. Grade: A-
The secondary did a good job clearing up many of the issues it had last week's loss, helping to allow Purdue to complete just 50% of its passes. The coverage was tighter, the communication was better and the tackling was less passive that it was against Michigan State. The coverage was a little lax at times in the fourth quarter, but it was still a big improvement from the previous game.
Shaun Prater led the secondary with six tackles, including a sack, forced a fumble and broke up two passes. Jordan Bernstine and Micah Hyde both finished with five stops, and though Tanner Miller only had one tackle, he was able to come away with two interceptions, giving him three this season. B.J. Lowery also had just one tackle but made it count with a sack in the fourth quarter. Grade: A-
Overall Defensive Grade: A- Things got a little interesting at the end of the game, but overall, the defense turned in one of its strongest performances of the season. After struggling last week in a loss, Iowa held Purdue to 282 yards of total offense, forced three turnovers and came up with five sacks, the most for a Hawkeye defense since a 12-0 win over Minnesota in 2009.
Special Teams
Despite making a couple of big plays, it was a so-so performance for the special teams, though it was an improvement from last week. Senior Kyle Steinbrecher came up with a blocked punt in the second quarter, giving Iowa its first blocked punt since the season-opener in 2010. Collin Sleeper recovered the punt to give the Hawkeyes great field position. Iowa also forced a fumble on a Purdue kick return, although like the blocked punt, the offense wasn't able to take advantage of the miscue. Though the average of 33.2-yard per punt wasn't great, Eric Guthrie did a good job punting in windy conditions, placing two of his four punts inside the 20. Bernstine did a decent job handling the kick returns, though he didn't break any big runs.
Though the conditions were less than ideal for a kicker, Mike Meyer had a bit of a rough day. He drilled a 38-yard field goal attempt but also missed a 34-yarder off the upright, the third time he has missed one off the upright this season. It was also Meyer's fourth miss in his last six attempts, although one of those came on a blocked kick. He also struggled on kickoffs, averaging just 55.7 yards per kick. Hyde let a punt get by him instead of catching it and allowed it to roll downfield for an additional 10-15 yards, something he has done many times this season, and the kick coverage unit was spotty at times. Grade: C
Overall Grade: B It wasn't always pretty but Iowa came back from a tough loss last week to walk away from West Lafayette with a much needed road victory. The offense was effective, if not a bit sloppy at times, and the defense had arguably its best outing of the Big Ten season as the Hawkeyes head into a pivotal final game of the regular season that could go a long way in determining the bowl pecking order.
Stat of the Game: +3
After losing the turnover battle last week, Iowa got back into the positive side of the column by forcing five Purdue turnovers (two picks, two fumble recoveries and a recovery of a blocked punt).


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NewswireToday - /newswire/ -
Stratford Upon Avon, Warwickshire, United Kingdom, 2008/06/16 - With call centre attrition rates booming in locations from Belfast to Bangalore. Cyber City Teleservices buck the trend and reward their employees with more than 5 years employment.
As part of its ongoing series looking at successful local businesses, The Sun Star Pampanga, a leading newspaper daily serving the Philippines, published a piece focusing on the tremendous employee loyalty for a Clark Freeport Zone-headquartered company, Cyber City Teleservices.
A global call centre with clients throughout the world, Cyber City Teleservices boasts a wide array of multi award-winning employee programs, along with an acclaimed on-site University that trains its employees for growth careers with the company.
At a time when most call centres can hardly cite employees that have stayed with their companies for more than 90 days, Cyber City Teleservices is proud of our workforce many of them who have served with our company for more than 5 years, said Linda Chando, who heads Cyber Citys day to day operations.
Said the Sun Star article:
The tradition goes on as Cyber City Teleservices commends its 176 regular employees for their continuous five-year service with the company. During the recognition ceremony for its Batch 2001 employees, Cyber City president Jonathan Rosenberg handed over a plaque of loyalty and appreciation to the honorees. Representatives from the group of awardees expressed their gratitude for the company's unwavering support in their personal and professional growth. They shared testimonies and memories of how they grew, enjoyed and viewed the company as their second home and family. Most of these employees now occupy managerial and supervisory positions at Cyber City. As a pioneer call centre in the country, Cyber City maintains its culture of appreciation and keeps its tradition of recognizing employees with five years of continuous service with the company. Two years ago, Cyber City celebrated a similar event, which included 97 employees hired in year 2000. Cyber City will be celebrating its 10th anniversary in the Philippines next summer.
Written by / Agency / Source: Cyber City Teleservices
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