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Sample records for highly thermogenic tissue

  1. Effects of Diets Differing in Composition of 18-C Fatty Acids on Adipose Tissue Thermogenic Gene Expression in Mice Fed High-Fat Diets

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    Sunhye Shin

    2018-02-01

    Full Text Available Dietary fatty acids play important roles in the regulation of fat accumulation or metabolic phenotype of adipocytes, either as brown or beige fat. However, a systematic comparison of effects of diets with different composition of 18-C fatty acids on browning/beiging phenotype has not been done. In this study, we compared the effects of different dietary fats, rich in specific 18-carbon fatty acids, on thermogenesis and lipid metabolism. Male C57BL/6 mice were fed a control diet containing 5.6% kcal fat from lard and 4.4% kcal fat from soybean oil (CON or high-fat diets (HFD containing 25% kcal from lard and 20% kcal fat from shea butter (stearic acid-rich fat; SHB, olive oil (oleic acid-rich oil; OO, safflower oil (linoleic acid-rich oil; SFO, or soybean oil (mixed oleic, linoleic, and α-linolenic acids; SBO ad libitum for 12 weeks, with or without a terminal 4-h norepinephrine (NE treatment. When compared to SHB, feeding OO, SFO, and SBO resulted in lower body weight gain. The OO fed group had the highest thermogenesis level, which resulted in lower body fat accumulation and improved glucose and lipid metabolism. Feeding SFO downregulated expression of lipid oxidation-related genes and upregulated expression of lipogenic genes, perhaps due to its high n-6:n-3 ratio. In general, HFD-feeding downregulated Ucp1 expression in both subcutaneous and epididymal white adipose tissue, and suppressed NE-induced Pgc1a expression in brown adipose tissue. These results suggest that the position of double bonds in dietary fatty acids, as well as the quantity of dietary fat, may have a significant effect on the regulation of oxidative and thermogenic conditions in vivo.

  2. Lipolytic and thermogenic depletion of adipose tissue in cancer cachexia.

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    Tsoli, Maria; Swarbrick, Michael M; Robertson, Graham R

    2016-06-01

    Although muscle wasting is the obvious manifestation of cancer cachexia that impacts on patient quality of life, the loss of lipid reserves and metabolic imbalance in adipose tissue also contribute to the devastating impact of cachexia. Depletion of fat depots in cancer patients is more pronounced than loss of muscle and often precedes, or even occurs in the absence of, reduced lean body mass. Rapid mobilisation of triglycerides stored within adipocytes to supply the body with fatty acids in periods of high-energy demand is normally mediated through a well-defined process of lipolysis involving the lipases ATGL, HSL and MGL. Studies into how these lipases contribute to fat loss in cancer cachexia have revealed the prominent role for ATGL in initiating lipolysis during adipose tissue atrophy, together with links between tumour-derived factors and the signalling pathways that control lipid flux within fat cells. The recent findings of increased thermogenesis in brown fat during cancer cachexia indicate that metabolically active adipose tissue contributes to the imbalance in energy homeostasis involved in catabolic wasting. Such energetically futile use of fatty acids liberated from adipose tissue to generate heat represents a maladaptive response in conjunction with anorexia experienced by cancer patients. As IL-6 release by tumours provokes lipolysis and activates the thermogenic programme in brown fat, this review explores the overlap in dysregulated metabolic processes due to inflammatory mediators in cancer cachexia and other disease states characterised by elevated cytokines such as obesity and diabetes. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

  3. Leanness of Lou/C rats does not require higher thermogenic capacity of brown adipose tissue.

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    Belouze, Maud; Sibille, Brigitte; Rey, Benjamin; Roussel, Damien; Romestaing, Caroline; Teulier, Loïc; Baetz, Delphine; Koubi, Harry; Servais, Stéphane; Duchamp, Claude

    2011-10-24

    Lou/C rats, an inbred strain of Wistar origin, remain lean throughout life and therefore represent a remarkable model of obesity resistance. To date, the exact mechanisms responsible for the leanness of Lou/C rats remain unknown. The aim of the present study was to investigate whether the leanness of Lou/C rats relies on increased thermogenic capacities in brown adipose tissue (BAT). Results showed that although daily energy expenditure was higher in Lou/C than in Wistar rats, BAT thermogenic capacity was not enhanced in Lou/C rats kept at thermoneutrality as demonstrated by reduced thermogenic response to norepinephrine in vivo, similar oxidative activity of BAT isolated mitochondria in vitro, similar levels of UCP1 mRNA and lower abundance of UCP1 protein in interscapular BAT depots. Relative abundance of β3-adrenergic receptor mRNA was lower in Lou/C BAT while that of GLUT4, FABP or CPT1 was not altered. Activity-related energy expenditure was however considerably increased at thermoneutrality as Lou/C rats demonstrated an impressively high spontaneous running activity in voluntary running wheels. Prolonged cold-exposure (4 °C) depressed the spontaneous running activity of Lou/C rats while BAT thermogenic capacity was increased as reflected by rises in BAT mass, oxidative activity and UCP1 expression. It is concluded that the leanness of Lou/C rats cannot be ascribed to higher thermogenic capacity of brown fat but rather to, at least in part, increased locomotor activity. BAT is not deficient in this rat strain as it can be stimulated by cold exposure when locomotor activity is reduced suggesting some substitution between these thermogenic processes. Copyright © 2011 Elsevier Inc. All rights reserved.

  4. Brown Adipose Tissue Thermogenic Capacity Is Regulated by Elovl6

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    Chong Yew Tan

    2015-12-01

    Full Text Available Although many transcriptional pathways regulating BAT have been identified, the role of lipid biosynthetic enzymes in thermogenesis has been less investigated. Whereas cold exposure causes changes in the fatty acid composition of BAT, the functional consequences of this remains relatively unexplored. In this study, we demonstrate that the enzyme Elongation of Very Long Chain fatty acids 6 (Elovl6 is necessary for the thermogenic action of BAT. Elovl6 is responsible for converting C16 non-essential fatty acids into C18 species. Loss of Elovl6 does not modulate traditional BAT markers; instead, it causes reduced expression of mitochondrial electron transport chain components and lower BAT thermogenic capacity. The reduction in BAT activity appears to be counteracted by increased beiging of scWAT. When beige fat is disabled by thermoneutrality or aging, Elovl6 KO mice gain weight and have increased scWAT mass and impaired carbohydrate metabolism. Overall, our study suggests fatty acid chain length is important for BAT function.

  5. Thermogenic profiling using magnetic resonance imaging of dermal and other adipose tissues

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    Kasza, Ildiko; Hernando, Diego; Roldán-Alzate, Alejandro; Alexander, Caroline M.; Reeder, Scott B.

    2016-01-01

    Dermal white adipose tissue (dWAT) was recently recognized for its potential to modify whole body metabolism. Here, we show that dWAT can be quantified using a high-resolution, fat-specific magnetic resonance imaging (MRI) technique. Noninvasive MRI has been used to describe adipocyte depots for many years; the MRI technique we describe uses an advanced fat-specific method to measure the thickness of dWAT, together with the total volume of WAT and the relative activation/fat depletion of brown adipose tissues (BAT). Since skin-embedded adipocytes may provide natural insulation, they provide an important counterpoint to the activation of thermogenic brown and beige adipose tissues, whereby these distinct depots are functionally interrelated and require simultaneous assay. This method was validated using characterized mouse cohorts of a lipodystrophic, dWAT-deficient strain (syndecan-1 KO) and 2 obese models (diet-induced obese mice and genetically obese animals, ob/ob). Using a preliminary cohort of normal human subjects, we found the thickness of skin-associated fat varied 8-fold, from 0.13–1.10 cm; on average, this depot is calculated to weigh 8.8 kg. PMID:27668285

  6. Thermogenic response to epinephrine in the forearm and abdominal subcutaneous adipose tissue

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    Simonsen, L; Bülow, J; Madsen, Jan Lysgård

    1992-01-01

    was infused at rates of 0.2 and 0.4 nmol.kg-1 x min-1. Whole body resting energy expenditure was 4.36 +/- 0.56 (SD) kJ/min. Energy expenditure increased to 5.14 +/- 0.74 and 5.46 +/- 0.79 kJ/min, respectively (P exchange ratio was 0.80 +/- 0......Whole body energy expenditure, thermogenic and metabolic changes in the forearm, and intercellular glucose concentrations in subcutaneous adipose tissue on the abdomen determined by microdialysis were measured during epinephrine infusion in healthy subjects. After a control period, epinephrine...

  7. PACAP is essential for the adaptive thermogenic response of brown adipose tissue to cold exposure.

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    Diané, Abdoulaye; Nikolic, Nikolina; Rudecki, Alexander P; King, Shannon M; Bowie, Drew J; Gray, Sarah L

    2014-09-01

    Pituitary adenylate cyclase-activating polypeptide (PACAP) is a widely distributed neuropeptide that acts as a neurotransmitter, neuromodulator, neurotropic factor, neuroprotectant, secretagogue, and neurohormone. Owing to its pleiotropic biological actions, knockout of Pacap (Adcyap1) has been shown to induce several abnormalities in mice such as impaired thermoregulation. However, the underlying physiological and molecular mechanisms remain unclear. A previous report has shown that cold-exposed Pacap null mice cannot supply appropriate levels of norepinephrine (NE) to brown adipocytes. Therefore, we hypothesized that exogenous NE would rescue the impaired thermogenic response of Pacap null mice during cold exposure. We compared the adaptive thermogenic capacity of Pacap(-/-) to Pacap(+/+) mice in response to NE when housed at room temperature (24 °C) and after a 3.5-week cold exposure (4 °C). Biochemical parameters, expression of thermogenic genes, and morphological properties of brown adipose tissue (BAT) and white adipose tissue (WAT) were also characterized. Results showed that there was a significant effect of temperature, but no effect of genotype, on the resting metabolic rate in conscious, unrestrained mice. However, the normal cold-induced increase in the basal metabolic rate and NE-induced increase in thermogenesis were severely blunted in cold-exposed Pacap(-/-) mice. These changes were associated with altered substrate utilization, reduced β3-adrenergic receptor (β3-Ar (Adrb3)) and hormone-sensitive lipase (Hsl (Lipe)) gene expression, and increased fibroblast growth factor 2 (Fgf2) gene expression in BAT. Interestingly, Pacap(-/-) mice had depleted WAT depots, associated with upregulated uncoupling protein 1 expression in inguinal WATs. These results suggest that the impairment of adaptive thermogenesis in Pacap null mice cannot be rescued by exogenous NE perhaps in part due to decreased β3-Ar-mediated BAT activation. © 2014 Society for

  8. Thermogenic response to epinephrine in the forearm and abdominal subcutaneous adipose tissue

    DEFF Research Database (Denmark)

    Simonsen, L; Bülow, J; Madsen, J

    1992-01-01

    Whole body energy expenditure, thermogenic and metabolic changes in the forearm, and intercellular glucose concentrations in subcutaneous adipose tissue on the abdomen determined by microdialysis were measured during epinephrine infusion in healthy subjects. After a control period, epinephrine......-1 and increased to 0.586 +/- 0.445 and 0.760 +/- 0.534 mumol.100 g-1 x min-1 (P subcutaneous adipose tissue on the abdomen was equal to the arterial concentration in the basal period but did not increase as much during infusion of epinephrine......, indicating glucose uptake in adipose tissue in this condition. If it is assumed that forearm skeletal muscle is representative for the average skeletal muscle, it can be calculated that on average 40% of the enhanced whole body oxygen uptake induced by infusion of epinephrine is taking place in skeletal...

  9. Thermoneutrality decreases thermogenic program and promotes adiposity in high-fat diet-fed mice.

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    Cui, Xin; Nguyen, Ngoc Ly T; Zarebidaki, Eleen; Cao, Qiang; Li, Fenfen; Zha, Lin; Bartness, Timothy; Shi, Hang; Xue, Bingzhong

    2016-05-01

    Brown/beige adipocytes are therapeutic targets to combat obesity due to their abilities to dissipate energy through adaptive thermogenesis. Most studies investigating induction of brown/beige adipocytes were conducted in cold condition (e.g., 4°C); much is unknown about how the thermogenic program of brown/beige adipocytes is regulated in thermoneutral condition (e.g., 30°C), which is within the thermal comfort zone of human dwellings in daily life. Therefore, this study aims to characterize the thermogenic program of brown/beige adipocytes in mice housed under ambient (22°C) versus thermoneutral condition (30°C). Male mice raised at 22°C or 30°C were fed either chow diet or high-fat (HF) diet for 20 weeks. Despite less food intake, chow-fed mice housed at 30°C remained the same body weight compared to mice at 22°C. However, these thermoneutrally housed mice displayed a decrease in the expression of thermogenic program in both brown and white fat depots with larger adipocytes. When pair-fed with chow diet, thermoneutrally housed mice showed an increase in body weight. Moreover, thermoneutrality increased body weight of mice fed with HF diet. This was associated with decreased expression of the thermogenic program in both brown and white fat depots of the thermoneutrally housed mice. The downregulation of the thermogenic program might have resulted from decreased sympathetic drive in the thermoneutrally housed mice evident by decreased expression of tyrosine hydroxylase expression and norepinephrine turnover in both brown and white fat depots. Our data demonstrate that thermoneutrality may negatively regulate the thermogenic program and sympathetic drive, leading to increased adiposity in mice. © 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

  10. Insulin resistance is not associated with thermogenic effect of a high-fat meal in obese children

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    Chan, Jeremy; Lomenick, Jefferson P.; Buchowski, Maciej S.; Shoemaker, Ashley H.

    2014-01-01

    In adults, insulin resistance may decrease thermogenic effect of food, contributing to weight gain. We aimed to determine the effect of insulin resistance on energy expenditure in children with long-standing obesity. We hypothesized that thermogenic effect of food would decrease with increasing insulin resistance. Energy expenditure was measured using whole room indirect calorimetry in obese children 7 – 18 years old. Participants were fed a high-fat meal with energy content equal to 35% of m...

  11. AT1 receptor antagonist induces thermogenic beige adipocytes in the inguinal white adipose tissue of obese mice.

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    Graus-Nunes, Francielle; Rachid, Tamiris Lima; de Oliveira Santos, Felipe; Barbosa-da-Silva, Sandra; Souza-Mello, Vanessa

    2017-03-01

    To evaluate whether losartan is able to induce beige adipocytes formation, focusing on the thermogenic gene expression and adipocyte remodeling in the subcutaneous white adipose tissue of diet-induced obese mice. Male C57BL/6 mice received a control diet (10% energy as lipids) or a high-fat diet (50% energy as lipids) for 10 weeks, followed by a 5-week treatment with losartan: control group, control-losartan group (10 mg/Kg/day), high-fat group and high-fat-losartan group (10 mg/Kg/day). Biochemical, morphometrical, stereological and molecular approaches were used to evaluate the outcomes. The high-fat diet elicited overweight, insulin resistance and adipocyte hypertrophy in the high-fat group, all of which losartan rescued in the high-fat-losartan group. These effects comply with the induction of beige adipocytes within the inguinal fat pads in high-fat-losartan group as they exhibited the greatest energy expenditure among the groups along with the presence uncoupling protein 1 positive multilocular adipocytes with enhanced peroxisome proliferator-activated receptor gamma coactivator 1-alpha and PR domain containing 16 mRNA levels, indicating a significant potential for mitochondrial biogenesis and adaptive thermogenesis. Our results show compelling evidence that losartan countered diet-induced obesity in mice by enhancing energy expenditure through beige adipocytes induction. Reduced body mass, increased insulin sensitivity, decreased adipocyte size and marked expression of uncoupling protein 1 by ectopic multilocular adipocytes support these findings. The use of losartan as a coadjutant medicine to tackle obesity and its related disorders merits further investigation.

  12. Lipocalin 2, a Regulator of Retinoid Homeostasis and Retinoid-mediated Thermogenic Activation in Adipose Tissue*

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    Guo, Hong; Foncea, Rocio; O'Byrne, Sheila M.; Jiang, Hongfeng; Zhang, Yuanyuan; Deis, Jessica A.; Blaner, William S.; Bernlohr, David A.; Chen, Xiaoli

    2016-01-01

    We have recently characterized the role of lipocalin 2 (Lcn2) as a new adipose-derived cytokine in the regulation of adaptive thermogenesis via a non-adrenergic pathway. Herein, we explored a potential non-adrenergic mechanism by which Lcn2 regulates thermogenesis and lipid metabolism. We found that Lcn2 is a retinoic acid target gene, and retinoic acid concurrently stimulated UCP1 and Lcn2 expression in adipocytes. Lcn2 KO mice exhibited a blunted effect of all-trans-retinoic acid (ATRA) on body weight and fat mass, lipid metabolism, and retinoic acid signaling pathway activation in adipose tissue under the high fat diet-induced obese condition. We further demonstrated that Lcn2 is required for the full action of ATRA on the induction of UCP1 and PGC-1α expression in brown adipocytes and the restoration of cold intolerance in Lcn2 KO mice. Interestingly, we discovered that Lcn2 KO mice have decreased levels of retinoic acid and retinol in adipose tissue. The protein levels of STRA6 responsible for retinol uptake were significantly decreased in adipose tissue. The retinol transporter RBP4 was increased in adipose tissue but decreased in the circulation, suggesting the impairment of RBP4 secretion in Lcn2 KO adipose tissue. Moreover, Lcn2 deficiency abolished the ATRA effect on RBP4 expression in adipocytes. All the data suggest that the decreased retinoid level and action are associated with impaired retinol transport and storage in adipose tissue in Lcn2 KO mice. We conclude that Lcn2 plays a critical role in regulating metabolic homeostasis of retinoids and retinoid-mediated thermogenesis in adipose tissue. PMID:27008859

  13. Lipocalin 2, a Regulator of Retinoid Homeostasis and Retinoid-mediated Thermogenic Activation in Adipose Tissue.

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    Guo, Hong; Foncea, Rocio; O'Byrne, Sheila M; Jiang, Hongfeng; Zhang, Yuanyuan; Deis, Jessica A; Blaner, William S; Bernlohr, David A; Chen, Xiaoli

    2016-05-20

    We have recently characterized the role of lipocalin 2 (Lcn2) as a new adipose-derived cytokine in the regulation of adaptive thermogenesis via a non-adrenergic pathway. Herein, we explored a potential non-adrenergic mechanism by which Lcn2 regulates thermogenesis and lipid metabolism. We found that Lcn2 is a retinoic acid target gene, and retinoic acid concurrently stimulated UCP1 and Lcn2 expression in adipocytes. Lcn2 KO mice exhibited a blunted effect of all-trans-retinoic acid (ATRA) on body weight and fat mass, lipid metabolism, and retinoic acid signaling pathway activation in adipose tissue under the high fat diet-induced obese condition. We further demonstrated that Lcn2 is required for the full action of ATRA on the induction of UCP1 and PGC-1α expression in brown adipocytes and the restoration of cold intolerance in Lcn2 KO mice. Interestingly, we discovered that Lcn2 KO mice have decreased levels of retinoic acid and retinol in adipose tissue. The protein levels of STRA6 responsible for retinol uptake were significantly decreased in adipose tissue. The retinol transporter RBP4 was increased in adipose tissue but decreased in the circulation, suggesting the impairment of RBP4 secretion in Lcn2 KO adipose tissue. Moreover, Lcn2 deficiency abolished the ATRA effect on RBP4 expression in adipocytes. All the data suggest that the decreased retinoid level and action are associated with impaired retinol transport and storage in adipose tissue in Lcn2 KO mice. We conclude that Lcn2 plays a critical role in regulating metabolic homeostasis of retinoids and retinoid-mediated thermogenesis in adipose tissue. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Insulin resistance is not associated with thermogenic effect of a high-fat meal in obese children.

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    Chan, Jeremy; Lomenick, Jefferson P; Buchowski, Maciej S; Shoemaker, Ashley H

    2014-06-01

    In adults, insulin resistance may decrease the thermogenic effect of food, contributing to weight gain. We aimed to determine the effect of insulin resistance on energy expenditure in children with long-standing obesity. We hypothesized that thermogenic effect of food would decrease with increasing insulin resistance. Energy expenditure was measured using whole room indirect calorimetry in obese children 7 to 18 years old. Participants were fed a high-fat meal with energy content equal to 35% of measured resting energy expenditure. Thermogenic effect of food was measured for 180 minutes posttest meal and expressed as a percent of calories consumed. Body composition was assessed using whole-body dual-energy x-ray absorptiometry. Fasting glucose, insulin, and hemoglobin A1C were measured. Complete data were available for 25 children (median age, 12.1 years; 52% male). As expected, a significant decrease in resting energy expenditure was observed with increasing Tanner stage (P = .02 by Kruskal-Wallis test). Insulin sensitivity, as determined by homeostasis model assessment index equation, did not significantly affect resting energy expenditure (P = .3) or thermogenic effect of food (P = .7) after adjustment for Tanner stage. In conclusion, our study did not find an association between insulin resistance and energy expenditure in obese children. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Metabolic consequences of the presence or absence of the thermogenic capacity of brown adipose tissue in mice (and probably in humans).

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    Cannon, B; Nedergaard, J

    2010-10-01

    Only with the development of the uncoupling protein 1 (UCP1)-ablated mouse has it become possible to strictly delineate the physiological significance of the thermogenic capacity of brown adipose tissue. Considering the presence of active brown adipose tissue in adult humans, these insights may have direct human implications. In addition to classical nonshivering thermogenesis, all adaptive adrenergic thermogeneses, including diet-induced thermogenesis, is fully dependent on brown adipocyte activity. Any weight-reducing effect of β(3)-adrenergic agonists is fully dependent on UCP1 activity, as is any weight-reducing effect of leptin (in excess of its effect on reduction of food intake). Consequently, in the absence of the thermogenic activity of brown adipose tissue, obesity develops spontaneously. The ability of brown adipose tissue to contribute to glucose disposal is also mainly related to thermogenic activity. However, basal metabolic rate, cold-induced thermogenesis, acute cold tolerance, fevers, nonadaptive adrenergic thermogenesis and processes such as angiogenesis in brown adipose tissue itself are not dependent on UCP1 activity. Whereas it is likely that these conclusions are also qualitatively valid for adult humans, the quantitative significance of brown adipose tissue for human metabolism--and the metabolic consequences for a single individual possessing more or less brown adipose tissue--awaits clarification.

  16. Tea catechin and caffeine activate brown adipose tissue and increase cold-induced thermogenic capacity in humans.

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    Yoneshiro, Takeshi; Matsushita, Mami; Hibi, Masanobu; Tone, Hiroshi; Takeshita, Masao; Yasunaga, Koichi; Katsuragi, Yoshihisa; Kameya, Toshimitsu; Sugie, Hiroki; Saito, Masayuki

    2017-04-01

    Background: The thermogenic effects of green tea catechin have been repeatedly reported, but their mechanisms are poorly understood.Objective: The aim of this study was to investigate the acute and chronic effects of catechin on brown adipose tissue (BAT), a site specialized for nonshivering thermogenesis, in humans.Design: Fifteen healthy male volunteers underwent fluorodeoxyglucose-positron emission tomography to assess BAT activity. To examine the acute catechin effect, whole-body energy expenditure (EE) after a single oral ingestion of a beverage containing 615 mg catechin and 77 mg caffeine (catechin beverage) was measured. Next, to investigate the chronic catechin effects, 10 men with low BAT activity were enrolled. Before and after ingestion of the catechin beverage 2 times/d for 5 wk, cold-induced thermogenesis (CIT) after 2 h of cold exposure at 19°C, which is proportional to BAT activity, was examined. Both the acute and chronic trials were single-blinded, randomized, placebo-controlled, season-matched crossover studies.Results: A single ingestion of the catechin beverage increased EE in 9 subjects who had metabolically active BAT (mean ± SEM: +15.24 ± 1.48 kcal, P caffeine produced a smaller and comparative EE response in the 2 subject groups. Multivariate regression analysis revealed a significant interaction between BAT and catechin on EE (β = 0.496, P = 0.003). Daily ingestion of the catechin beverage elevated mean ± SEM CIT (from 92.0 ± 26.5 to 197.9 ± 27.7 kcal/d; P = 0.009), whereas the placebo beverage did not change it.Conclusion: Orally ingested tea catechin with caffeine acutely increases EE associated with increased BAT activity and chronically elevates nonshivering CIT, probably because of the recruitment of BAT, in humans. These trials were registered at www.umin.ac.jp/ctr/ as UMIN000016361. © 2017 American Society for Nutrition.

  17. Thermoneutrality decreases thermogenic program and promotes adiposity in high?fat diet?fed mice

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    Cui, Xin; Ngoc Ly T. Nguyen; Zarebidaki, Eleen; Cao, Qiang; Li, Fenfen; Zha, Lin; Bartness, Timothy; Shi, Hang; Xue, Bingzhong

    2016-01-01

    Abstract Brown/beige adipocytes are therapeutic targets to combat obesity due to their abilities to dissipate energy through adaptive thermogenesis. Most studies investigating induction of brown/beige adipocytes were conducted in cold condition (e.g., 4?C); much is unknown about how the thermogenic program of brown/beige adipocytes is regulated in thermoneutral condition (e.g., 30?C), which is within the thermal comfort zone of human dwellings in daily life. Therefore, this study aims to char...

  18. Thermogenic mechanisms and their hormonal regulation.

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    Silva, J Enrique

    2006-04-01

    Increased heat generation from biological processes is inherent to homeothermy. Homeothermic species produce more heat from sustaining a more active metabolism as well as from reducing fuel efficiency. This article reviews the mechanisms used by homeothermic species to generate more heat and their regulation largely by thyroid hormone (TH) and the sympathetic nervous system (SNS). Thermogenic mechanisms antecede homeothermy, but in homeothermic species they are activated and regulated. Some of these mechanisms increase ATP utilization (same amount of heat per ATP), whereas others increase the heat resulting from aerobic ATP synthesis (more heat per ATP). Among the former, ATP utilization in the maintenance of ionic gradient through membranes seems quantitatively more important, particularly in birds. Regulated reduction of the proton-motive force to produce heat, originally believed specific to brown adipose tissue, is indeed an ancient thermogenic mechanism. A regulated proton leak has been described in the mitochondria of several tissues, but its precise mechanism remains undefined. This leak is more active in homeothermic species and is regulated by TH, explaining a significant fraction of its thermogenic effect. Homeothermic species generate additional heat, in a facultative manner, when obligatory thermogenesis and heat-saving mechanisms become limiting. Facultative thermogenesis is activated by the SNS but is modulated by TH. The type II iodothyronine deiodinase plays a critical role in modulating the amount of the active TH, T(3), in BAT, thereby modulating the responses to SNS. Other hormones affect thermogenesis in an indirect or permissive manner, providing fuel and modulating thermogenesis depending on food availability, but they do not seem to have a primary role in temperature homeostasis. Thermogenesis has a very high energy cost. Cold adaptation and food availability may have been conflicting selection pressures accounting for the variability of

  19. Deficiency of Interleukin-15 Confers Resistance to Obesity by Diminishing Inflammation and Enhancing the Thermogenic Function of Adipose Tissues.

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    Gregory Lacraz

    Full Text Available IL-15 is an inflammatory cytokine secreted by many cell types. IL-15 is also produced during physical exercise by skeletal muscle and has been reported to reduce weight gain in mice. Contrarily, our findings on IL-15 knockout (KO mice indicate that IL-15 promotes obesity. The aim of this study is to investigate the mechanisms underlying the pro-obesity role of IL-15 in adipose tissues.Control and IL-15 KO mice were maintained on high fat diet (HFD or normal control diet. After 16 weeks, body weight, adipose tissue and skeletal mass, serum lipid levels and gene/protein expression in the adipose tissues were evaluated. The effect of IL-15 on thermogenesis and oxygen consumption was also studied in primary cultures of adipocytes differentiated from mouse preadipocyte and human stem cells.Our results show that IL-15 deficiency prevents diet-induced weight gain and accumulation of lipids in visceral and subcutaneous white and brown adipose tissues. Gene expression analysis also revealed elevated expression of genes associated with adaptive thermogenesis in the brown and subcutaneous adipose tissues of IL-15 KO mice. Accordingly, oxygen consumption was increased in the brown adipocytes from IL-15 KO mice. In addition, IL-15 KO mice showed decreased expression of pro-inflammatory mediators in their adipose tissues.Absence of IL-15 results in decreased accumulation of fat in the white adipose tissues and increased lipid utilization via adaptive thermogenesis. IL-15 also promotes inflammation in adipose tissues that could sustain chronic inflammation leading to obesity-associated metabolic syndrome.

  20. Unlock the thermogenic potential of adipose tissue: pharmacological modulation and implications for treatment of diabetes and obesity

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    Xiao-Rong ePeng

    2015-11-01

    Full Text Available Brown adipose tissue (BAT is considered an interesting target organ for the treatment of metabolic disease due to its high metabolic capacity. Non-shivering thermogenesis, once activated, can lead to enhanced partitioning and oxidation of fuels in adipose tissues, and reduce the burden of glucose and lipids on other metabolic organs such as liver, pancreas and skeletal muscle. Sustained long-term activation of BAT may also lead to meaningful bodyweight loss. In this review, we discuss three different drug classes (the thiazolidinedione (TZD class of PPAR agonists, 3 adrenergic receptor agonists and fibroblast growth factor 21 (FGF21 analogues that have been proposed to regulate BAT and beige recruitment or activation, or both, and which have been tested in both rodent and human. The learnings from these classes suggest that restoration of functional BAT and beige mass as well as improved activation might be required to fully realize the metabolic potential of these tissues. Whether this can be achieved without the undesired cardiovascular side-effects exhibited by the TZD PPAR agonists and 3 adrenergic receptor agonists remains to be resolved.

  1. A thermogenic secondary sexual character in male sea lamprey

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    Chung-Davidson, Yu-Wen; Priess, M. Cody; Yeh, Chu-Yin; Brant, Cory O.; Johnson, Nicholas S.; Li, Ke; Nanlohy, Kaben G.; Bryan, Mara B.; Brown, C. Titus; Choi, Jongeun; Li, Weiming

    2013-01-01

    Secondary sexual characters in animals are exaggerated ornaments or weapons for intrasexual competition. Unexpectedly, we found that a male secondary sexual character in sea lamprey (Petromyzon marinus ) is a thermogenic adipose tissue that instantly increases its heat production during sexual encounters. This secondary sexual character, developed in front of the anterior dorsal fin of mature males, is a swollen dorsal ridge known as the ‘rope’ tissue. It contains nerve bundles, multivacuolar adipocytes and interstitial cells packed with small lipid droplets and mitochondria with dense and highly organized cristae. The fatty acid composition of the rope tissue is rich in unsaturated fatty acids. The cytochrome c oxidase activity is high but the ATP concentration is very low in the mitochondria of the rope tissue compared with those of the gill and muscle tissues. The rope tissue temperature immediately rose up to 0.3°C when the male encountered a conspecific. Mature males generated more heat in the rope and muscle tissues when presented with a mature female than when presented with a male (paired t-test, P-3 more heat than the muscle in 10 min. Transcriptome analyses revealed that genes involved in fat cell differentiation are upregulated whereas those involved in oxidative-phosphorylation-coupled ATP synthesis are downregulated in the rope tissue compared with the gill and muscle tissues. Sexually mature male sea lamprey possess the only known thermogenic secondary sexual character that shows differential heat generation toward individual conspecifics.

  2. Exendin-4 improves thermogenic capacity by regulating fat metabolism on brown adipose tissue in mice with diet-induced obesity.

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    Wei, Qiong; Li, Ling; Chen, Ji-an; Wang, Shao-hua; Sun, Zi-lin

    2015-01-01

    This study aimed to investigate the benefits of exendin-4 treatment on brown adipose tissue (BAT) in C57BL/6J mice with high-fat diet (HFD)-induced obesity. We examined the effects of exendin-4 on body adiposity and the level of genes associated with adipogenesis, glucose/lipid uptake, lipolysis, and thermogenesis in mice with diet-induced obesity. Exendin-4 treatment deceased body weight, serum-free fatty acid, and triglyceride levels in HFD-induced obese C57BL/6J mice. Exendin-4 treatment increased the expression of genes associated with adipogenesis, glucose/lipid uptake, lipolysis, and thermogenesis in BAT. Compared with HFD-fed mice, exendin-4 treatment also exhibited elevated energy expenditure and reduced respiratory quotient, but showed similar food intake and locomotor activity. Exendin-4 treatment reduced high-fat-induced obesity by decreasing adiposity and increasing thermogenesis. This result suggests that GLP-1 agonist may be a new approach to combat obesity by shifting the energy balance from obesogenesis to thermogenesis. © 2015 by the Association of Clinical Scientists, Inc.

  3. Clonal analyses and gene profiling identify genetic biomarkers of the thermogenic potential of human brown and white preadipocytes.

    Science.gov (United States)

    Xue, Ruidan; Lynes, Matthew D; Dreyfuss, Jonathan M; Shamsi, Farnaz; Schulz, Tim J; Zhang, Hongbin; Huang, Tian Lian; Townsend, Kristy L; Li, Yiming; Takahashi, Hirokazu; Weiner, Lauren S; White, Andrew P; Lynes, Maureen S; Rubin, Lee L; Goodyear, Laurie J; Cypess, Aaron M; Tseng, Yu-Hua

    2015-07-01

    Targeting brown adipose tissue (BAT) content or activity has therapeutic potential for treating obesity and the metabolic syndrome by increasing energy expenditure. However, both inter- and intra-individual differences contribute to heterogeneity in human BAT and potentially to differential thermogenic capacity in human populations. Here we generated clones of brown and white preadipocytes from human neck fat and characterized their adipogenic and thermogenic differentiation. We combined an uncoupling protein 1 (UCP1) reporter system and expression profiling to define novel sets of gene signatures in human preadipocytes that could predict the thermogenic potential of the cells once they were maturated. Knocking out the positive UCP1 regulators, PREX1 and EDNRB, in brown preadipocytes using CRISPR-Cas9 markedly abolished the high level of UCP1 in brown adipocytes differentiated from the preadipocytes. Finally, we were able to prospectively isolate adipose progenitors with great thermogenic potential using the cell surface marker CD29. These data provide new insights into the cellular heterogeneity in human fat and offer potential biomarkers for identifying thermogenically competent preadipocytes.

  4. The suppression of ghrelin signaling mitigates age-associated thermogenic impairment

    Science.gov (United States)

    Bongmba, Odelia Y. N.; Ma, Xiaojun; Zhu, Xiongwei; Sheikh-Hamad, David; Sun, Yuxiang

    2014-01-01

    Aging is associated with severe thermogenic impairment, which contributes to obesity and diabetes in aging. We previously reported that ablation of the ghrelin receptor, growth hormone secretagogue receptor (GHS-R), attenuates age-associated obesity and insulin resistance. Ghrelin and obestatin are derived from the same preproghrelin gene. Here we showed that in brown adipocytes, ghrelin decreases the expression of thermogenic regulator but obestatin increases it, thus showing the opposite effects. We also found that during aging, plasma ghrelin and GHS-R expression in brown adipose tissue (BAT) are increased, but plasma obestatin is unchanged. Increased plasma ghrelin and unchanged obestatin during aging may lead to an imbalance of thermogenic regulation, which may in turn exacerbate thermogenic impairment in aging. Moreover, we found that GHS-R ablation activates thermogenic signaling, enhances insulin activation, increases mitochondrial biogenesis, and improves mitochondrial dynamics of BAT. In addition, we detected increased norepinephrine in the circulation, and observed that GHS-R knockdown in brown adipocytes directly stimulates thermogenic activity, suggesting that GHS-R regulates thermogenesis via both central and peripheral mechanisms. Collectively, our studies demonstrate that ghrelin signaling is an important thermogenic regulator in aging. Antagonists of GHS-R may serve as unique anti-obesity agents, combating obesity by activating thermogenesis. PMID:25543537

  5. Moderate alcohol intake induces thermogenic brown/beige adipocyte formationviaelevating retinoic acid signaling.

    Science.gov (United States)

    Wang, Bo; Wang, Zhixiu; de Avila, Jeanene M; Zhu, Mei-Jun; Zhang, Faya; Gomez, Noe Alberto; Zhao, Liang; Tian, Qiyu; Zhao, Junxing; Maricelli, Joseph; Zhang, Hui; Rodgers, Buel D; Du, Min

    2017-10-01

    Clinically, low and moderate alcohol intake improves human health with protection against metabolic syndromes, including type 2 diabetes; however, mechanisms that are associated with these effects remain to be elucidated. The aims of this study were to investigate the effects of moderate alcohol intake on thermogenic brown/beige adipocyte formation and glucose and lipid homeostasis, as well as the involvement of retinoic acid (RA) signaling in the entire process. C57BL6 male mice were supplemented with 8% (w/v) alcohol in water for 1 or 4 mo. Alcohol intake prevented body weight gain, induced the formation of uncoupling protein 1-positive beige adipocytes in white adipose tissue, and increased thermogenesis in mice, which is associated with decreased serum glucose and triacylglycerol levels. Mechanistically, alcohol intake increased RA levels in serum and adipose tissue, which was associated with increased expression of aldehyde dehydrogenase family 1 subfamily A1 ( Aldh1a1 ). When RA receptor-α signaling was conditionally blocked in platelet-derived growth factor receptor-α-positive adipose progenitors, the effects of alcohol on beige adipogenesis were largely abolished. Finally, moderate alcohol prevented high-fat diet-induced obesity and metabolic dysfunction. In conclusion, moderate alcohol intake induces thermogenic brown/beige adipocyte formation and promotes glucose and lipid oxidation via elevation of RA signaling.-Wang, B., Wang, Z., de Avila, J. M., Zhu, M.-J., Zhang, F., Gomez, N. A., Zhao, L., Tian, Q., Zhao, J., Maricelli, J., Zhang, H., Rodgers, B. D., Du, M. Moderate alcohol intake induces thermogenic brown/beige adipocyte formation via elevating retinoic acid signaling. © FASEB.

  6. Leptin "gates" thermogenic action of thyrotropin-releasing hormone in the hindbrain.

    Science.gov (United States)

    Rogers, Richard C; Barnes, Maria J; Hermann, Gerlinda E

    2009-10-27

    Leptin, acting as a measure of metabolic fuel availability, exerts a powerful permissive influence on neurogenic thermogenesis. During starvation and an absence of leptin, animals cannot produce thermogenic reactions to cold stress. However, thermogenesis is rescued by restoring leptin. We have previously observed (Hermann, G.E., Barnes, M.J., Rogers, R.C., 2006. Leptin and thyrotropin-releasing hormone: cooperative action in the hindbrain to activate brown adipose thermogenesis. Brain Res. 1117, 118-124.) a highly cooperative interaction between leptin and thyrotropin-releasing hormone [TRH] to activate hindbrain generated thermogenic responses. Specifically, exposure to both leptin and TRH elicited a 3.5 degrees C increase in brown adipose tissue [BAT] thermogenesis, while leptin alone did not evoke any change, and TRH alone caused only approximately 1 degrees C increase. The present study shows that the leptin-TRH synergy in controlling brown adipose [BAT] thermogenesis is order-specific and dependent on the feeding status of the animal. That is, fourth ventricular [4V] application of leptin to the food-deprived animal, before TRH injection, yields a substantial increase in BAT; while the reverse order yields a significantly smaller effect. If the animal were fed within minutes of anesthesia, then exogenous leptin was not necessary for TRH to yield a large increase in BAT temperature. The leptin-TRH synergy was uncoupled by pretreatment with the phosphoinositol-tris phosphate kinase [PI3K] inhibitor, wortmannin and the Src-SH2 antagonist, PP2. The TRH transduction mechanism utilizes phospholipase C [PLC] potently regulated by the SH2 site. Previous work in culture systems suggests that the product of PI3K activity [PIP3] potently upregulates PLC by activating the SH2 domain of the PLC complex. Perhaps leptin "gates" the thermogenic action of TRH in the hindbrain by invoking this same mechanism.

  7. Thermogenic characterization of ghrelin receptor null mice.

    Science.gov (United States)

    Lin, Ligen; Sun, Yuxiang

    2012-01-01

    Ghrelin is the only known circulating orexigenic hormone that increases food intake and promotes adiposity, and these physiological functions of ghrelin are mediated through its receptor growth hormone secretagogue receptor (GHS-R). Ghrelin/GHS-R signaling plays a crucial role in energy homeostasis. Old GHS-R null mice exhibit a healthy phenotype-lean and insulin sensitive. Interestingly, the GHS-R null mice have increased energy expenditure, yet exhibit no difference in food intake or locomotor activity compared to wild-type mice. We have found that GHS-R is expressed in brown adipose tissue (BAT) of old mice. Ablation of GHS-R attenuates age-associated decline in thermogenesis, exhibiting a higher core body temperature. Indeed, the BAT of old GHS-R null mice reveals enhanced thermogenic capacity, which is consistent with the gene expression profile of increases in glucose/lipid uptake, lipogenesis, and lipolysis in BAT. The data collectively suggest that ghrelin/GHS-R signaling has important roles in thermogenesis. The recent discovery that BAT also regulates energy homeostasis in adult humans makes the BAT a new antiobesity target. Understanding the roles and molecular mechanisms of ghrelin/GHS-R in thermogenesis is of great significance. GHS-R antagonists might be a novel means of combating obesity by shifting adiposity balance from obesogenesis to thermogenesis. Copyright © 2012 Elsevier Inc. All rights reserved.

  8. Characterizing thermogenic coalbed gas from Polish coals of different ranks by hydrous pyrolysis

    Science.gov (United States)

    Kotarba, M.J.; Lewan, M.D.

    2004-01-01

    To provide a better characterization of origin and volume of thermogenic gas generation from coals, hydrous pyrolysis experiments were conducted at 360??C for 72 h on Polish coals ranging in rank from lignite (0.3% R r) to semi-anthracite (2.0% Rr). Under these conditions, the lignites attained a medium-volatile bituminous rank (1.5% Rr), high-volatile bituminous coals attained a low-volatile bituminous rank (1.7% Rr), and the semi-anthracite obtained an anthracite rank (4.0% R r). Hydrous pyrolysis of a coal, irrespective of rank, provides a diagnostic ??13C value for its thermogenic hydrocarbon gases. This value can be used quantitatively to interpret mixing of indigenous thermogenic gas with microbial methane or exogenous thermogenic gas from other sources. Thermogenic methane quantities range from 20 dm3/kg of lignite (0.3% Rr) to 0.35 dm3/kg of semi-anthracite (2.0% Rr). At a vitrinite reflectance of 1.7% Rr, approximately 75% of the maximum potential for a coal to generate thermogenic methane has been expended. At a vitrinite reflectance of 1.7% Rr, more than 90% of the maximum potential for a coal to generate CO2 has been expended. Assuming that these quantities of generated CO2 remain associated with a sourcing coal bed as uplift or erosion provide conditions conducive for microbial methanogenesis, the resulting quantities of microbial methane generated by complete CO2 reduction can exceed the quantities of thermogenic methane generated from the same coal bed by a factor of 2-5. ?? 2004 Elsevier Ltd. All rights reserved.

  9. Genome-Wide Insights into the Development and Function of Thermogenic Adipocytes

    DEFF Research Database (Denmark)

    Loft, Anne; Forss, Isabel; Mandrup, Susanne

    2017-01-01

    Brown and brown-like adipocytes are specialized adipocytes with a high capacity to convert metabolic energy to heat. This function is not only eminent in supporting organismal thermogenesis, but may also have potential in the fight against obesity. The latter has spurred a massive interest...... in understanding the development and regulation of these thermogenic adipocytes. Here, we review how genome-wide studies based on next-generation sequencing have provided insight into how the chromatin and transcriptional landscapes are established in thermogenic adipocytes and how thermogenic signals can change...... the genomic programming of white adipocytes. Furthermore, we discuss how the integration of genomic data can be used to discover novel transcriptional pathways that may be modulated as part of therapeutic strategies for the treatment of obesity....

  10. Thermogenic Wet Gas in Immature Caprock Sections: Leakage or Generation.

    Science.gov (United States)

    Abrakasa, Selegha; Beka, Francis; Ndukauba, Egesi

    2017-04-01

    Gas geochemistry, an aspect of Petroleum Geoscience is a growing science, various concepts has been used to evaluation potential source rock for shale gas while in conventional petroleum exploration similar concepts have been used to determine potential productive formation for liquid hydrocarbons. Prior to the present times, headspace gas data had been used to recognize by pass pays, serve as indicators of petroleum accumulations, evaluate maturity and productive capacity of corresponding formations, evaluate the maturity and source of gas accumulations. Integrating studies in bid to achieve high degree of accuracy, data on direct hydrocarbon indicators (DHIs) such as oil stains, oil shows and seeps have been employed. Currently popular among professionals is the use of gas clouds on seismic cross sections. In contemporary times, advancement in gas geochemistry has witnessed the application of concepts on headspace gas to expound the efficiency of petroleum caprocks whose major role is to foster accumulation and preservation. This enables extricating potential leakage mechanism via caprock reservoir interface and unravel its corresponding migrational pathways. In this study thermogenic wet gas has been used as a dependable tool for delineating caprock leakage by discriminating migrant from indigenous hydrocarbons in caprock rock sections overlying the reservoirs. The thermogenic gas profile in corroboration with the thermogenic signature and maturity data were used. Summary statistics indicates that 60% of the 50 wells studied has wet gas up to 500m above the reservoir-caprock interface and 10% of the leaking wells are fracture prone leakage.The amount of wet gas ranges of up to 200,000 ppm in the caprock sections, this indicates pervasive leakage. Log view plots were modelled using Schlumbergers' Techlog, while descriptive lithologies were modeled using Zetawares' genesis.

  11. Ablations of ghrelin and ghrelin receptor exhibit differential metabolic phenotypes and thermogenic capacity during aging.

    Science.gov (United States)

    Ma, Xiaojun; Lin, Ligen; Qin, Guijun; Lu, Xinping; Fiorotto, Marta; Dixit, Vishwa D; Sun, Yuxiang

    2011-01-26

    Obesity is a hallmark of aging in many Western societies, and is a precursor to numerous serious age-related diseases. Ghrelin (Ghrl), via its receptor (growth hormone secretagogue receptor, GHS-R), is shown to stimulate GH secretion and appetite. Surprisingly, our previous studies showed that Ghrl(-/-) mice have impaired thermoregulatory responses to cold and fasting stresses, while Ghsr(-/-) mice are adaptive. To elucidate the mechanism, we analyzed the complete metabolic profiles of younger (3-4 months) and older (10-12 months) Ghrl(-/-) and Ghsr(-/-) mice. Food intake and locomotor activity were comparable for both null mice and their wild-type (WT) counterparts, regardless of age. There was also no difference in body composition between younger null mice and their WT counterparts. As the WT mice aged, as expected, the fat/lean ratio increased and energy expenditure (EE) decreased. Remarkably, however, older Ghsr(-/-) mice exhibited reduced fat/lean ratio and increased EE when compared to older WT mice, thus retaining a youthful lean and high EE phenotype; in comparison, there was no significant difference with EE in Ghrl(-/-) mice. In line with the EE data, the thermogenic regulator, uncoupling protein 1 (UCP1), was significantly up-regulated in brown adipose tissue (BAT) of Ghsr(-/-) mice, but not in Ghrl(-/-) mice. Our data therefore suggest that GHS-R ablation activates adaptive thermogenic function(s) in BAT and increases EE, thereby enabling the retention of a lean phenotype. This is the first direct evidence that the ghrelin signaling pathway regulates fat-burning BAT to affect energy balance during aging. This regulation is likely mediated through an as-yet-unidentified new ligand of GHS-R.

  12. Ablations of ghrelin and ghrelin receptor exhibit differential metabolic phenotypes and thermogenic capacity during aging.

    Directory of Open Access Journals (Sweden)

    Xiaojun Ma

    2011-01-01

    Full Text Available Obesity is a hallmark of aging in many Western societies, and is a precursor to numerous serious age-related diseases. Ghrelin (Ghrl, via its receptor (growth hormone secretagogue receptor, GHS-R, is shown to stimulate GH secretion and appetite. Surprisingly, our previous studies showed that Ghrl(-/- mice have impaired thermoregulatory responses to cold and fasting stresses, while Ghsr(-/- mice are adaptive.To elucidate the mechanism, we analyzed the complete metabolic profiles of younger (3-4 months and older (10-12 months Ghrl(-/- and Ghsr(-/- mice. Food intake and locomotor activity were comparable for both null mice and their wild-type (WT counterparts, regardless of age. There was also no difference in body composition between younger null mice and their WT counterparts. As the WT mice aged, as expected, the fat/lean ratio increased and energy expenditure (EE decreased. Remarkably, however, older Ghsr(-/- mice exhibited reduced fat/lean ratio and increased EE when compared to older WT mice, thus retaining a youthful lean and high EE phenotype; in comparison, there was no significant difference with EE in Ghrl(-/- mice. In line with the EE data, the thermogenic regulator, uncoupling protein 1 (UCP1, was significantly up-regulated in brown adipose tissue (BAT of Ghsr(-/- mice, but not in Ghrl(-/- mice.Our data therefore suggest that GHS-R ablation activates adaptive thermogenic function(s in BAT and increases EE, thereby enabling the retention of a lean phenotype. This is the first direct evidence that the ghrelin signaling pathway regulates fat-burning BAT to affect energy balance during aging. This regulation is likely mediated through an as-yet-unidentified new ligand of GHS-R.

  13. A new, highly selective murine peroxisome proliferator-activated receptor δ agonist increases responsiveness to thermogenic stimuli and glucose uptake in skeletal muscle in obese mice.

    Science.gov (United States)

    Ngala, R A; Stocker, C J; Roy, A G; Hislop, D; Wargent, E; Bell, R; Hassall, D G; Harling, J D; Billin, A N; Willson, T M; Arch, J R S; Cawthorne, M A

    2011-05-01

    We investigated how GW800644, the first pharmacologically selective murine peroxisome proliferator-activated receptor δ (PPARδ) agonist, affects energy balance, glucose homeostasis and fuel utilization by muscle in obese mice. Potencies were determined in transactivation assays. Oral glucose tolerance was determined after 14 and 22 days' administration (10 mg/kg body weight, twice daily) to Lep(ob)/Lep(ob) mice. Food intake and energy expenditure were measured during a 26-day experiment, and plasma metabolites and 2-deoxyglucose uptake in vivo at termination. Palmitate oxidation and 2-deoxyglucose uptake by isolated soleus muscles were measured after 14 (in lean and obese mice) and 26 days. GW800644 activated murine PPARδ (EC(50) 2 nM), but caused little to no activation of PPARα or PPARγ up to 10 µM. It did not increase liver weight. GW800644 reduced food intake and body weight in obese mice after 8 days. It did not affect resting energy expenditure, but, compared to pair-fed mice, it increased the response to a β(3)-adrenoceptor agonist. It improved glucose tolerance. GW800644, but not pair-feeding, reduced plasma glucose, insulin and triglyceride concentrations. It increased 2-deoxyglucose uptake in vivo in adipose tissue, soleus muscle, heart, brain and liver, and doubled 2-deoxyglucose uptake and palmitate oxidation in isolated soleus muscle from obese but not lean mice. PPARδ agonism reduced food intake and independently elicited metabolic effects that included increased responsiveness to β(3)-adrenoceptor stimulation, increased glucose utilization and fat oxidation in soleus muscle of Lep(ob)/Lep(ob) but not lean mice and increased glucose utilization in vivo in Lep(ob)/Lep(ob) mice. © 2011 Blackwell Publishing Ltd.

  14. Pro-opiomelanocortin modulates the thermogenic and physical activity responses to high-fat feeding and markedly influences dietary fat preference.

    Science.gov (United States)

    Tung, Y C Loraine; Rimmington, Debra; O'Rahilly, Stephen; Coll, Anthony P

    2007-11-01

    Complete proopiomelanocortin (POMC) deficiency causes a human syndrome of hypoadrenalism, altered skin and hair pigmentation, and severe hyperphagic obesity. Heterozygote carriers of nonsense mutations are strongly predisposed to obesity. Pomc(+/-) mice have normal body weight on a chow diet but increase food intake and become more obese than wild-type littermates when placed on a high-fat diet. To further explore the mechanisms whereby dietary fat interacts with Pomc genotype to produce obesity, we examined Pomc-null, Pomc(+/-), and wild-type mice for changes in the components of energy balance in response to provision of a high-fat diet and macronutrient preference when presented with a selection of dietary choices. In contrast to wild-type mice, Pomc null mice did not increase their resting energy expenditure or their spontaneous physical activity when given a high-fat diet. Pomc(+/-) mice increased resting energy expenditure similarly to wild types, but their increase in physical activity was significantly less than that seen in wild-type mice. In two independent experimental tests of macronutrient preference, Pomc genotype was a strong predictor of dietary fat preference with Pomc null animals choosing to eat approximately twice as much fat, but similar amounts of carbohydrate and protein, as wild-type animals. Pomc(+/-) mice showed an intermediate response. In summary, POMC-derived peptides have influences on multiple aspects of the organism's response to the presentation of high-fat diet. This includes a major influence, readily discernible even in heterozygote animals, on the dietary preference for fat.

  15. Central serotonergic neurons activate and recruit thermogenic brown and beige fat and regulate glucose and lipid homeostasis

    DEFF Research Database (Denmark)

    McGlashon, Jacob M; Gorecki, Michelle C; Kozlowski, Amanda E

    2015-01-01

    Thermogenic brown and beige adipocytes convert chemical energy to heat by metabolizing glucose and lipids. Serotonin (5-HT) neurons in the CNS are essential for thermoregulation and accordingly may control metabolic activity of thermogenic fat. To test this, we generated mice in which the human...... diphtheria toxin receptor (DTR) was selectively expressed in central 5-HT neurons. Treatment with diphtheria toxin (DT) eliminated 5-HT neurons and caused loss of thermoregulation, brown adipose tissue (BAT) steatosis, and a >50% decrease in uncoupling protein 1 (Ucp1) expression in BAT and inguinal white...

  16. Effects of nutrient intake on sympathoadrenal activity and thermogenic mechanisms

    DEFF Research Database (Denmark)

    Astrup, A V; Christensen, N J; Simonsen, L

    1990-01-01

    the physiological threshold for thermogenic effect. The target is mainly skeletal muscle where thermogenesis is stimulated via beta 2-adrenoceptors. Also the basal metabolic rate and the thermogenic responses to cold and heat exposure, mental stress and exercise, have facultative components. Inhibition...

  17. The thermogenic center in social wasps.

    Science.gov (United States)

    Ishay, Jacob S; Plotkin, Marian; Ermakov, Natalya Y; Volynchik, Stanislav; Barkay, Zahava; Bergman, David J

    2006-01-01

    In the social wasps Vespa orientalis and Paravespula germanica (Hymenoptera, Vespinae), a thermogenic center has been found in the dorsal part of the first thoracic segment. The temperature in this region of the prothorax is higher by 6-9 degrees C than that at the tip of the abdomen, and this in actively flying hornets outside the nest (workers, males or queens) as well as in hornets inside the nest that attend to the brood in the combs. On viewing the region from the outside, one discerns a canal or rather a fissure in the cuticle, which commences at the center of the dorsal surface of the prothorax and extends till the mesothorax. Thus the length of this canal or fissure is approximately 5-7 mm and it is seen to contain numerous thin hairs whose shape varies from that of the hairs alongside the structure. Beneath the cuticle in this region there are dorsoventral as well as longitudinal muscles in abundance, much the same as the musculature in the remaining thoracic segments (i.e. the meso- and metathorax), which activate the two pairs of wings. The canal-bearing segment is of course devoid of wings, and its dorsoventral muscles are attached to the cuticle, which in this region resembles a bowl harboring several layers of epithelium that boasts numerous butterfly-shaped tracheal branches. Additionally there are layers that display lymph-filled spaces and also perforated layers and depressions, and beneath all these is a lace-like layer that also coats the cuticle's hollows. Underneath the cuticle proper, there are numerous large mitochondria and tracheae, which occupy a considerable part of the cuticular epithelium surface. These abundant mitochondria are, most probably, the main element of heat production in the thermogenic center.

  18. Thermogenic capacity is antagonistically regulated in classical brown and white subcutaneous fat depots by high fat diet and endurance training in rats: impact on whole-body energy expenditure

    National Research Council Canada - National Science Library

    Wu, Michelle V; Bikopoulos, George; Hung, Steven; Ceddia, Rolando B

    2014-01-01

    ...) and subcutaneous inguinal (SC Ing) white adipose tissue (WAT) and how it affects whole-body energy expenditure in sedentary and endurance-trained rats fed ad libitum either low fat or high fat (HF) diets...

  19. Obesity-promoting and anti-thermogenic effects of neutrophil gelatinase-associated lipocalin in mice.

    Science.gov (United States)

    Ishii, Akira; Katsuura, Goro; Imamaki, Hirotaka; Kimura, Hiroyuki; Mori, Keita P; Kuwabara, Takashige; Kasahara, Masato; Yokoi, Hideki; Ohinata, Kousaku; Kawanishi, Tomoko; Tsuchida, Junichi; Nakamoto, Yuji; Nakao, Kazuwa; Yanagita, Motoko; Mukoyama, Masashi; Mori, Kiyoshi

    2017-11-14

    Neutrophil gelatinase-associated lipocalin (NGAL, lipocalin 2 or LCN2) is an iron carrier protein whose circulating level is increased by kidney injury, bacterial infection and obesity, but its metabolic consequence remains elusive. To study physiological role of LCN2 in energy homeostasis, we challenged female Lcn2 knockout (KO) and wild-type (WT) mice with high fat diet (HFD) or cold exposure. Under normal diet, physical constitutions of Lcn2 KO and WT mice were indistinguishable. During HFD treatment, Lcn2 KO mice exhibited larger brown adipose tissues (BAT), consumed more oxygen, ate more food and gained less body weights as compared to WT mice. When exposed to 4 °C, KO mice showed higher body temperature and more intense 18F-fluorodeoxyglucose uptake in BAT, which were cancelled by β3 adrenergic receptor blocker or iron-loaded (but not iron-free) LCN2 administration. These findings suggest that circulating LCN2 possesses obesity-promoting and anti-thermogenic effects through inhibition of BAT activity in an iron-dependent manner.

  20. Increased locomotor and thermogenic activity in mice with targeted ablation of the GHRH gene.

    Science.gov (United States)

    Leone, Sheila; Chiavaroli, Annalisa; Shohreh, Rugia; Ferrante, Claudio; Ricciuti, Adriana; Manippa, Fabio; Recinella, Lucia; Di Nisio, Chiara; Orlando, Giustino; Salvatori, Roberto; Vacca, Michele; Brunetti, Luigi

    2015-04-01

    Growth hormone (GH) deficiency (GHD) leads to growth failure and changes in body composition, including increased fat accumulation and reduced lean body mass in both humans and rodents. The aim of this study was to examine the factors that contribute to energy imbalance in the GH releasing hormone knock out (GHRHKO) mice, a well established model of GHD. We evaluated food intake (of standard laboratory chow), total body weight (TBW), locomotor activity, body temperature and interscapular brown adipose tissue (BAT) weight in 8 adult male mice homozygous for the GHRHKO allele (-/-) and 8 heterozygous (+/-) animals as controls. The gene expression of uncoupling protein-1 (UCP-1) in BAT and the levels of norepinephrine (NE), dopamine (DA), and serotonin (5-hydroxytryptamine, 5-HT) in the ventral striatum were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) and high performance liquid chromatography (HPLC) analysis, respectively. Throughout 2 months of observation -/- mice consumed approximately 40% more food (normalized to TBW; Pfood intake observed in GHRH ablated animals is associated with increased locomotor and thermogenic activity. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Adipocyte lipid synthesis coupled to neuronal control of thermogenic programming

    Directory of Open Access Journals (Sweden)

    Adilson Guilherme

    2017-08-01

    Conclusions: These results demonstrate that downregulation of fatty acid synthesis via FASN depletion in white adipocytes of mature mice can stimulate neuronal signaling to control thermogenic programming in iWAT.

  2. Thermogenic effect of glucose in hypothyroid subjects.

    Science.gov (United States)

    Kozacz, Agnieszka; Grunt, Paulina; Steczkowska, Marta; Mikulski, Tomasz; Dąbrowski, Jan; Górecka, Monika; Sanocka, Urszula; Ziemba, Andrzej Wojciech

    2014-01-01

    The importance of thyroid hormone, catecholamines, and insulin in modification of the thermogenic effect of glucose (TEG) was examined in 34 healthy and 32 hypothyroid subjects. We calculated the energy expenditure at rest and during oral glucose tolerance test. Blood samples for determinations of glucose, plasma insulin, adrenaline (A), and noradrenaline (NA) were collected. It was found that TEG was lower in hypothyroid than in control group (19.68 ± 3.90 versus 55.40 ± 7.32 kJ, resp., P < 0.0004). Mean values of glucose and insulin areas under the curve were higher in women with hypothyroidism than in control group (286.79 ± 23.65 versus 188.41 ± 15.84 mmol/L·min, P < 0.003 and 7563.27 ± 863.65 versus 4987.72 ± 583.88 mU/L·min, P < 0.03 resp.). Maximal levels of catecholamines after glucose ingestion were higher in hypothyroid patients than in control subjects (Amax-0.69 ± 0.08 versus 0.30 ± 0.07 nmol/L, P < 0.0001, and NAmax-6.42 ± 0.86 versus 2.54 ± 0.30 nmol/L, P < 0.0002). It can be concluded that in hypothyroidism TEG and glucose tolerance are decreased while the adrenergic response to glucose administration is enhanced. Presumably, these changes are related to decreased insulin sensitivity and responsiveness to catecholamine action.

  3. Thermogenic Effect of Glucose in Hypothyroid Subjects

    Directory of Open Access Journals (Sweden)

    Agnieszka Kozacz

    2014-01-01

    Full Text Available The importance of thyroid hormone, catecholamines, and insulin in modification of the thermogenic effect of glucose (TEG was examined in 34 healthy and 32 hypothyroid subjects. We calculated the energy expenditure at rest and during oral glucose tolerance test. Blood samples for determinations of glucose, plasma insulin, adrenaline (A, and noradrenaline (NA were collected. It was found that TEG was lower in hypothyroid than in control group (19.68±3.90 versus 55.40±7.32 kJ, resp., P<0.0004. Mean values of glucose and insulin areas under the curve were higher in women with hypothyroidism than in control group (286.79±23.65 versus 188.41±15.84 mmol/L·min, P<0.003 and 7563.27±863.65 versus 4987.72±583.88 mU/L·min, P<0.03 resp.. Maximal levels of catecholamines after glucose ingestion were higher in hypothyroid patients than in control subjects (Amax—0.69±0.08 versus 0.30±0.07 nmol/L, P<0.0001, and NAmax—6.42±0.86 versus 2.54±0.30 nmol/L, P<0.0002. It can be concluded that in hypothyroidism TEG and glucose tolerance are decreased while the adrenergic response to glucose administration is enhanced. Presumably, these changes are related to decreased insulin sensitivity and responsiveness to catecholamine action.

  4. Anterograde transneuronal viral tract tracing reveals central sensory circuits from brown fat and sensory denervation alters its thermogenic responses

    OpenAIRE

    Vaughan, Cheryl H.; Bartness, Timothy J.

    2012-01-01

    Brown adipose tissue (BAT) thermogenic activity and growth are controlled by its sympathetic nervous system (SNS) innervation, but nerve fibers containing sensory-associated neuropeptides [substance P, calcitonin gene-related peptide (CGRP)] also suggest sensory innervation. The central nervous system (CNS) projections of BAT afferents are unknown. Therefore, we used the H129 strain of the herpes simplex virus-1 (HSV-1), an anterograde transneuronal viral tract tracer used to delineate sensor...

  5. Hypothalamic actions of tumor necrosis factor alpha provide the thermogenic core for the wastage syndrome in cachexia.

    Science.gov (United States)

    Arruda, Ana Paula; Milanski, Marciane; Romanatto, Talita; Solon, Carina; Coope, Andressa; Alberici, Luciane C; Festuccia, William T; Hirabara, Sandro M; Ropelle, Eduardo; Curi, Rui; Carvalheira, José B; Vercesi, Aníbal E; Velloso, Licio A

    2010-02-01

    TNFalpha is an important mediator of catabolism in cachexia. Most of its effects have been characterized in peripheral tissues, such as skeletal muscle and fat. However, by acting directly in the hypothalamus, TNFalpha can activate thermogenesis and modulate food intake. Here we show that high concentration TNFalpha in the hypothalamus leads to increased O(2) consumption/CO(2) production, increased body temperature, and reduced caloric intake, resulting in loss of body mass. Most of the thermogenic response is produced by beta 3-adrenergic signaling to the brown adipose tissue (BAT), leading to increased BAT relative mass, reduction in BAT lipid quantity, and increased BAT mitochondria density. The expression of proteins involved in BAT thermogenesis, such as beta 3-adrenergic receptor, peroxisomal proliferator-activated receptor-gamma coactivator-1 alpha, and uncoupling protein-1, are increased. In the hypothalamus, TNFalpha produces reductions in neuropeptide Y, agouti gene-related peptide, proopiomelanocortin, and melanin-concentrating hormone, and increases CRH and TRH. The activity of the AMP-activated protein kinase signaling pathway is also decreased in the hypothalamus of TNFalpha-treated rats. Upon intracerebroventricular infliximab treatment, tumor-bearing and septic rats present a significantly increased survival. In addition, the systemic inhibition of beta 3-adrenergic signaling results in a reduced body mass loss and increased survival in septic rats. These data suggest hypothalamic TNFalpha action to be important mediator of the wastage syndrome in cachexia.

  6. Hypothalamic ERK mediates the anorectic and thermogenic sympathetic effects of leptin.

    Science.gov (United States)

    Rahmouni, Kamal; Sigmund, Curt D; Haynes, William G; Mark, Allyn L

    2009-03-01

    Leptin is an adipocyte hormone that plays a major role in energy balance. Leptin receptors in the hypothalamus are known to signal via distinct mechanisms, including signal transducer and activator of transcription-3 (STAT3) and phosphoinositol-3 kinase (PI 3-kinase). Here, we tested the hypothesis that extracellular signal-regulated kinase (ERK) is mediating leptin action in the hypothalamus. Biochemical, pharmacological, and physiological approaches were combined to characterize leptin activation of ERK in the hypothalamus in rats. Leptin activates ERK1/2 in a receptor-mediated manner that involves JAK2. Leptin-induced ERK1/2 activation was restricted to the hypothalamic arcuate nucleus. Pharmacological blockade of hypothalamic ERK1/2 reverses the anorectic and weight-reducing effects of leptin. The pharmacological antagonists of ERK1/2 did not attenuate leptin-induced activation of STAT3 or PI 3-kinase. Blockade of ERK1/2 abolishes leptin-induced increases in sympathetic nerve traffic to thermogenic brown adipose tissue (BAT) but does not alter the stimulatory effects of leptin on sympathetic nerve activity to kidney, hindlimb, or adrenal gland. In contrast, blockade of PI 3-kinase prevents leptin-induced sympathetic activation to kidney but not to BAT, hindlimb, or adrenal gland. Our findings indicate that hypothalamic ERK plays a key role in the control of food intake, body weight, and thermogenic sympathetic outflow by leptin but does not participate in the cardiovascular and renal sympathetic actions of leptin.

  7. Pharmacological characterisation of the thermogenic effect of bupropion.

    Science.gov (United States)

    Liu, Yong-Ling; Connoley, Ian P; Heal, David J; Stock, Michael J

    2004-09-13

    The pharmacological mechanism of bupropion's thermogenic effect has been investigated in female Wistar rats by measuring oxygen consumption at thermoneutrality (29 degrees C). Bupropion (30 mg/kg) rapidly increased oxygen consumption (VO2) with a maximum effect at 30 min, and VO2 remained elevated throughout the 4-h experimental period. The nonselective 5-hydroxytryptamine (5-HT or serotonin) receptor antagonist, metergoline (1 mg/kg), and the alpha1-adrenoceptor antagonist, prazosin (1 mg/kg), had no effect on the VO2 response to bupropion, whereas the alpha2-adrenoceptor antagonist, RS79948 [(8aR, 12aS, 13aS)-5,8,8a,9,10,11,12,12a,13,13a-decahydro-3-methoxy-12-(ethylsulphonyl)-6H-isoquino[2,1-g][1,6]-naphthyridine hydrochloride] (1 mg/kg), potentiated the response. The VO2 response to bupropion during the first 60 min was significantly inhibited by a high dose of the nonselective beta-adrenoceptor antagonist, propranolol (20 mg/kg), but it had no effect at a low dose (1 mg/kg). Pretreatment with the dopamine D2/D1 receptor antagonist, (+)butaclamol (200 microg/kg), caused a partial, but significant, inhibition (Pfood intake in rats. Hence, in this species, its weight-reducing action predominantly results from thermogenesis mediated via activation of beta3-adrenergic and dopamine D2/D1 receptors. Because bupropion has also been reported not to alter food intake in the clinic, thermogenesis may also contribute to its antiobesity effect in man.

  8. Thermogenic effect and substrate oxidation of protein from animal and plant sources in adults

    Directory of Open Access Journals (Sweden)

    Ahmed Fahmy Arif Tsani

    2015-01-01

    Full Text Available Background: Changing nutrient source is one of the efforts to increase thermogenic effect (TEF which may be significant for body weight reduction. Objective: The aim of this study was to investigate the effects of high protein diets using animal (chicken and plant (tofu sources on the thermogenic effect (TEF and substrate oxidation. Method: Ten female adults (mean age 20.8+1.2 y participated in two isocaloric diet ingestions. Each meal provided 30% of the daily basal energy need (32/26/42% as protein/fat/carbohydrates, respectively. Postprandial energy expenditure was measured by indirect calorimetry. Results: There were no significant differences in TEF and substrate oxidation. The postprandial fat oxidation rate was higher than that at the preprandial state, while carbohydrate and protein oxidation rates were lower. Conclusion: No differences were observed in TEF and substrate oxidation in animal- and plant-based diets. A high protein diet could be beneficial for weight loss, but animal protein does not appear to offer superior benefits compared to plant protein.

  9. Second messenger signaling mechanisms of the brown adipocyte thermogenic program: an integrative perspective.

    Science.gov (United States)

    Shi, Fubiao; Collins, Sheila

    2017-09-26

    β-adrenergic receptors (βARs) are well established for conveying the signal from catecholamines to adipocytes. Acting through the second messenger cyclic adenosine monophosphate (cAMP) they stimulate lipolysis and also increase the activity of brown adipocytes and the 'browning' of adipocytes within white fat depots (so-called 'brite' or 'beige' adipocytes). Brown adipose tissue mitochondria are enriched with uncoupling protein 1 (UCP1), which is a regulated proton channel that allows the dissipation of chemical energy in the form of heat. The discovery of functional brown adipocytes in humans and inducible brown-like ('beige' or 'brite') adipocytes in rodents have suggested that recruitment and activation of these thermogenic adipocytes could be a promising strategy to increase energy expenditure for obesity therapy. More recently, the cardiac natriuretic peptides and their second messenger cyclic guanosine monophosphate (cGMP) have gained attention as a parallel signaling pathway in adipocytes, with some unique features. In this review, we begin with some important historical work that touches upon the regulation of brown adipocyte development and physiology. We then provide a synopsis of some recent advances in the signaling cascades from β-adrenergic agonists and natriuretic peptides to drive thermogenic gene expression in the adipocytes and how these two pathways converge at a number of unexpected points. Finally, moving from the physiologic hormonal signaling, we discuss yet another level of control downstream of these signals: the growing appreciation of the emerging roles of non-coding RNAs as important regulators of brown adipocyte formation and function. In this review, we discuss new developments in our understanding of the signaling mechanisms and factors including new secreted proteins and novel non-coding RNAs that control the function as well as the plasticity of the brown/beige adipose tissue as it responds to the energy needs and environmental

  10. The effect of chlorpyrifos on thermogenic capacity of bank voles selected for increased aerobic exercise metabolism.

    Science.gov (United States)

    Dheyongera, Geoffrey; Grzebyk, Katherine; Rudolf, Agata M; Sadowska, Edyta T; Koteja, Paweł

    2016-04-01

    Agro-chemicals potentially cause adverse effects in non-target organisms. The rate of animal energy metabolism can influence their susceptibility to pesticides by influencing food consumption, biotransformation and elimination rates of toxicants. We used experimental evolution to study the effects of inherent differences in energy metabolism rate and exposure to the organophosphate insecticide, chlorpyrifos (CPF) on thermogenic capacity in a wild rodent, the bank vole (Myodes = Clethrionomys glareolus). The voles were sampled from four replicate lines selected for high swim-induced aerobic metabolism (A) and four unselected control (C) lines. Thermogenic capacity, measured as the maximum cold-induced rate of oxygen consumption (VO2cold), was higher in the A - than C lines, and it decreased after continuous exposure to CPF via food or after a single dose administered via oral gavage, but only when measured shortly after exposure. VO2cold measured 24 h after repeated exposure was not affected. In addition, gavage with a single dose led to decreased food consumption and loss in body mass. Importantly, the adverse effects of CPF did not differ between the selected and control lines. Therefore, exposure to CPF has adverse effects on thermoregulatory performance and energy balance in this species. The effects are short-lived and their magnitude is not associated with the inherent level of energy metabolism. Even without severe symptoms of poisoning, fitness can be compromised under harsh environmental conditions, such as cold and wet weather. Copyright © 2016. Published by Elsevier Ltd.

  11. Optimized tissue heating by adopting high frequency electrotherapy

    Directory of Open Access Journals (Sweden)

    Jae-cheol Lee

    2015-11-01

    Full Text Available We have developed an electronics circuit that generates a high voltage with a frequency of 0.3–2 MHz to build an electro therapy system that can optimize tissue heating characteristics. These characteristics are used in medical applications. This paper is focused on the analysis of high frequency electro-therapy system to optimize tissue heating with the help of a high voltage pulse signal, which peak voltage is almost 2 kV. This optimized tissue heating between the inner tissue and the thermal distributions has examined in terms of frequency and voltage. The target tissue heating is composed of a single electrode in an experiment that has especially conducted to find the tissue heating characteristics. In the end, a new method for electro-therapy is developed, which is applicable to a specific tissue depth.

  12. Genetic disruption of uncoupling protein 1 in mice renders brown adipose tissue a significant source of FGF21 secretion

    Directory of Open Access Journals (Sweden)

    Susanne Keipert

    2015-07-01

    Conclusions: Here we show that the genetic ablation of UCP1 increases FGF21 gene expression in adipose tissue. The removal of adaptive nonshivering thermogenesis renders BAT a significant source of endogenous FGF21 under thermal stress. Thus, the thermogenic competence of BAT is not a requirement for FGF21 secretion. Notably, high endogenous FGF21 levels in UCP1-deficient models and subjects may confound pharmacological FGF21 treatments.

  13. Understanding the Biology of Thermogenic Fat: Is Browning A New Approach to the Treatment of Obesity?

    Science.gov (United States)

    Vargas-Castillo, Ariana; Fuentes-Romero, Rebeca; Rodriguez-Lopez, Leonardo A; Torres, Nimbe; Tovar, Armando R

    2017-07-01

    Obesity is characterized by an excess of white adipose tissue (WAT). Recent evidence has demonstrated that WAT can change its phenotype to a brown-like adipose tissue known as beige/brite adipose tissue. This transition is characterized by an increase in thermogenic capacity mediated by uncoupling protein 1 (UCP1). This browning process is a potential new target for treating obesity. The aim of this review is to integrate the different mechanisms by which beige/brite adipocytes are formed and to describe the physiological, pharmacological and nutritional inducers that can promote browning. An additional aim is to show evidence of how some of these inducers can be used as potential therapeutic agents against obesity and its comorbidities. This review shows the importance of brown and beige/brite adipose tissue and the mechanisms of their formation. Particularly, the two theories of beige/brite adipocyte origin are discussed: de novo differentiation and transdifferentiation. The gene markers that identify these types of adipocytes and the involvement of microRNAs in the epigenetic regulation of the browning process is also discussed. Additionally, we describe the transcriptional control of UCP1 expression by some of the inducers of browning. Furthermore, we describe in detail how some bioactive dietary compounds can induce browning and their subsequent beneficial health effects. The evidence suggests that browning is a new potential strategy for the treatment of obesity and obesity-associated metabolic disorders. Copyright © 2017 IMSS. Published by Elsevier Inc. All rights reserved.

  14. Sympathetically-mediated thermogenic response to food in rats.

    Science.gov (United States)

    Liverini, G; Iossa, S; Lionetti, L; Mollica, M P; Barletta, A

    1995-02-01

    The aim of this work was to assess the participation of the sympathetic nervous system in the thermogenic response to food in control and hyperphagic rats. Rats were fed either a control (CD) or energy dense (ED) diet. After 15 days, CD rats received a small (7 kJ) meal composed of either control or energy dense diet, while ED rats received a small meal composed of energy dense diet. The experiment was then repeated, with the exception that rats received a larger portion (35 kJ) of the test meal. The postprandial increase in oxygen consumption was measured for 30 min after the small meal and 90-180 min after the completion of the large meal. The measurements were made in saline-injected and propranolol-injected rats. ED rats exhibited hyperphagia as well as an increase of 32% in resting metabolic rate after a 16 h fast. The sympathetically-mediated postprandial increase in oxygen consumption was greater after an energy dense meal than after a control meal in CD rats, and was higher in ED rats than in CD rats fed an energy dense meal. It was concluded that the sympathetically-mediated increase in the thermogenic response to food, as well as the increase in fasting metabolic rate can help prevent obesity development in hyperphagic rats.

  15. Anterograde transneuronal viral tract tracing reveals central sensory circuits from brown fat and sensory denervation alters its thermogenic responses.

    Science.gov (United States)

    Vaughan, Cheryl H; Bartness, Timothy J

    2012-05-01

    Brown adipose tissue (BAT) thermogenic activity and growth are controlled by its sympathetic nervous system (SNS) innervation, but nerve fibers containing sensory-associated neuropeptides [substance P, calcitonin gene-related peptide (CGRP)] also suggest sensory innervation. The central nervous system (CNS) projections of BAT afferents are unknown. Therefore, we used the H129 strain of the herpes simplex virus-1 (HSV-1), an anterograde transneuronal viral tract tracer used to delineate sensory nerve circuits, to define these projections. HSV-1 was injected into interscapular BAT (IBAT) of Siberian hamsters and HSV-1 immunoreactivity (ir) was assessed 24, 48, 72, 96, and 114 h postinjection. The 96- and 114-h groups had the most HSV-1-ir neurons with marked infections in the hypothalamic paraventricular nucleus, periaqueductal gray, olivary areas, parabrachial nuclei, raphe nuclei, and reticular areas. These sites also are involved in sympathetic outflow to BAT suggesting possible BAT sensory-SNS thermogenesis feedback circuits. We tested the functional contribution of IBAT sensory innervation on thermogenic responses to an acute (24 h) cold exposure test by injecting the specific sensory nerve toxin capsaicin directly into IBAT pads and then measuring core (T(c)) and IBAT (T(IBAT)) temperature responses. CGRP content was significantly decreased in capsaicin-treated IBAT demonstrating successful sensory nerve destruction. T(IBAT) and T(c) were significantly decreased in capsaicin-treated hamsters compared with the saline controls at 2 h of cold exposure. Thus the central sensory circuits from IBAT have been delineated for the first time, and impairment of sensory feedback from BAT appears necessary for the appropriate, initial thermogenic response to acute cold exposure.

  16. The beta-adrenergic antagonist propranolol partly abolishes thermogenic response to bioactive food ingredients.

    Science.gov (United States)

    Belza, Anita; Gille, Maj-Britt; Schultz John, Sarah; Kondrup, Jens

    2009-08-01

    A combination of tyrosine, capsaicin, catechins, and caffeine has been shown to possess a thermogenic effect in humans. The present objective was to investigate whether the thermogenic response to the bioactive combination (BC) could be diminished or abolished by propranolol. Twenty-two men (age, 29.0 +/- 7.1 years; body mass index, 26.0 +/- 3.6 kg/m(2); mean +/- SD) participated in a 4-way, randomized, double-blind, placebo-controlled crossover study. The effect of the following was tested: (1) placebo, (2) BC, (3) BC + 5 mg propranolol, and (4) BC + 10 mg propranolol. Resting metabolic rate, respiratory quotient, and the thermogenic response were measured for 5 hours postintake. Systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate, and appetite ratings were assessed every half hour. The BC increased resting metabolic rate by 5% (73 [36; 110] kJ/5 h, mean [95% confidence interval], P thermogenic response by 50% compared with placebo (P thermogenic properties of BC. The 50% reduction of the thermogenic response by propranolol indicates that beta-adrenergic pathways are partly responsible for the thermogenic response.

  17. Thermogenic activity of UCP1 in human white fat-derived beige adipocytes.

    Science.gov (United States)

    Bartesaghi, Stefano; Hallen, Stefan; Huang, Li; Svensson, Per-Arne; Momo, Remi A; Wallin, Simonetta; Carlsson, Eva K; Forslöw, Anna; Seale, Patrick; Peng, Xiao-Rong

    2015-01-01

    Heat-producing beige/brite (brown-in-white) adipocytes in white adipose tissue have the potential to suppress metabolic disease in mice and hold great promise for the treatment of obesity and type 2 diabetes in humans. Here, we demonstrate that human adipose-derived stromal/progenitor cells (hASCs) from subcutaneous white adipose tissue can be efficiently converted into beige adipocytes. Upon pharmacological activation of peroxisome proliferator-activated receptor-γ, hASC-derived adipocytes activated beige fat-selective genes and a brown/beige fat-selective electron transport chain gene program. Importantly, hASC-derived beige fat cells displayed the bioenergetic characteristics of genuine brown fat cells, including a capacity for increased respiratory uncoupling in response to β-adrenergic agonists. Furthermore, knock-down experiments reveal that the thermogenic capacity of human beige fat cells was entirely dependent on the presence of Uncoupling protein 1. In summary, this study reveals that hASCs can be readily differentiated into beige adipocytes that, upon activation, undergo uncoupling protein 1-dependent thermogenesis.

  18. Basal metabolic rate, maximum thermogenic capacity and aerobic scope in rodents: interaction between environmental temperature and torpor use

    OpenAIRE

    Careau, Vincent

    2013-01-01

    When torpid animals arouse and warm up to restore normal body temperature (Tb), they produce heat at levels that can reach up to 10 times basal metabolic rate (BMR), close to the cold-induced summit metabolism (VO2-sum). Because torpor is an adaptation aimed at conserving energy over periods of low ambient temperature (Ta) and food availability, selective forces that have led to the evolution of torpor may have simultaneously favoured high thermogenic capacity (i.e. VO2-sum) relative to the m...

  19. Optical Histology: High-Resolution Visualization of Tissue Microvasculature

    Science.gov (United States)

    Moy, Austin Jing-Ming

    Mammalian tissue requires the delivery of nutrients, growth factors, and the exchange of oxygen and carbon dioxide gases to maintain normal function. These elements are delivered by the blood, which travels through the connected network of blood vessels, known as the vascular system. The vascular system consists of large feeder blood vessels (arteries and veins) that are connected to the small blood vessels (arterioles and venules), which in turn are connected to the capillaries that are directly connected to the tissue and facilitate gas exchange and nutrient delivery. These small blood vessels and capillaries make up an intricate but organized network of blood vessels that exist in all mammalian tissues known as the microvasculature and are very important in maintaining the health and proper function of mammalian tissue. Due to the importance of the microvasculature in tissue survival, disruption of the microvasculature typically leads to tissue dysfunction and tissue death. The most prevalent method to study the microvasculature is visualization. Immunohistochemistry (IHC) is the gold-standard method to visualize tissue microvasculature. IHC is very well-suited for highly detailed interrogation of the tissue microvasculature at the cellular level but is unwieldy and impractical for wide-field visualization of the tissue microvasculature. The objective my dissertation research was to develop a method to enable wide-field visualization of the microvasculature, while still retaining the high-resolution afforded by optical microscopy. My efforts led to the development of a technique dubbed "optical histology" that combines chemical and optical methods to enable high-resolution visualization of the microvasculature. The development of the technique first involved preliminary studies to quantify optical property changes in optically cleared tissues, followed by development and demonstration of the methodology. Using optical histology, I successfully obtained high

  20. The suppression of ghrelin signaling mitigates age-associated thermogenic impairment

    OpenAIRE

    Lin, Ligen; Lee, Jong Han; Bongmba, Odelia Y. N.; Ma, Xiaojun; Zhu, Xiongwei; Sheikh-Hamad, David; Sun, Yuxiang

    2014-01-01

    Aging is associated with severe thermogenic impairment, which contributes to obesity and diabetes in aging. We previously reported that ablation of the ghrelin receptor, growth hormone secretagogue receptor (GHS-R), attenuates age-associated obesity and insulin resistance. Ghrelin and obestatin are derived from the same preproghrelin gene. Here we showed that in brown adipocytes, ghrelin decreases the expression of thermogenic regulator but obestatin increases it, thus showing the opposite ef...

  1. A Review of Natural Stimulant and Non?stimulant Thermogenic Agents

    OpenAIRE

    Stohs, Sidney J.; Badmaev, Vladimir

    2016-01-01

    Obesity and overweight are major health issues. Exercise and calorie intake control are recognized as the primary mechanisms for addressing excess body weight. Naturally occurring thermogenic plant constituents offer adjunct means for assisting in weight management. The controlling mechanisms for thermogenesis offer many intervention points. Thermogenic agents can act through stimulation of the central nervous system with associated adverse cardiovascular effects and through metabolic mechani...

  2. [Protection of cadaver tissues exposed to high gamma radiation].

    Science.gov (United States)

    Matus-Jiménez, J; Flores-Fletes, J R; Carrillo, A

    2013-01-01

    Bone tissue is the most widely used tissue for the treatment of various conditions. As a result of this, allografts are used at an increasing frequency and processes for their harvest, preservation and sterilization have improved. The sterilization method that grants the greatest sterilization is high-dose gamma radiation, which destroys prions and any microorganism thus assuring that patients will not experience any infection. But given that radiation use has proven to deteriorate bone and tendon tissue, efforts have been made to protect the latter. One way to do this is a commercially available substance called Clearant. Studies conducted elsewhere have found that it does protect bone and tendon tissue. This study was therefore conducted with allograft samples exposed to high-dose radiation. Its purpose was to assess, with photon microscopy using various dyes and electron microscopy, the presence of color changes as well as the destruction of the anatomical structure. The same tissue was followed-up throughout the process until it was placed in the patient. The review found no structural changes in bone and tendon tissues exposed to high radiation doses (60 kilograys) when the Clearant process was used, and concluded that the former may be used safely in orthopedic or traumatologic diseases.

  3. A Review of Natural Stimulant and Non-stimulant Thermogenic Agents.

    Science.gov (United States)

    Stohs, Sidney J; Badmaev, Vladimir

    2016-05-01

    Obesity and overweight are major health issues. Exercise and calorie intake control are recognized as the primary mechanisms for addressing excess body weight. Naturally occurring thermogenic plant constituents offer adjunct means for assisting in weight management. The controlling mechanisms for thermogenesis offer many intervention points. Thermogenic agents can act through stimulation of the central nervous system with associated adverse cardiovascular effects and through metabolic mechanisms that are non-stimulatory or a combination thereof. Examples of stimulatory thermogenic agents that will be discussed include ephedrine and caffeine. Examples of non-stimulatory thermogenic agents include p-synephrine (bitter orange extract), capsaicin, forskolin (Coleus root extract), and chlorogenic acid (green coffee bean extract). Green tea is an example of a thermogenic with the potential to produce mild but clinically insignificant undesirable stimulatory effects. The use of the aforementioned thermogenic agents in combination with other extracts such as those derived from Salacia reticulata, Sesamum indicum, Lagerstroemia speciosa, Cissus quadrangularis, and Moringa olifera, as well as the use of the carotenoids as lutein and fucoxanthin, and flavonoids as naringin and hesperidin can further facilitate energy metabolism and weight management as well as sports performance without adverse side effects. © 2016 The Authors Phytotherapy Research published by John Wiley & Sons Ltd. © 2016 The Authors Phytotherapy Research published by John Wiley & Sons Ltd.

  4. A Review of Natural Stimulant and Non‐stimulant Thermogenic Agents

    Science.gov (United States)

    Badmaev, Vladimir

    2016-01-01

    Obesity and overweight are major health issues. Exercise and calorie intake control are recognized as the primary mechanisms for addressing excess body weight. Naturally occurring thermogenic plant constituents offer adjunct means for assisting in weight management. The controlling mechanisms for thermogenesis offer many intervention points. Thermogenic agents can act through stimulation of the central nervous system with associated adverse cardiovascular effects and through metabolic mechanisms that are non‐stimulatory or a combination thereof. Examples of stimulatory thermogenic agents that will be discussed include ephedrine and caffeine. Examples of non‐stimulatory thermogenic agents include p‐synephrine (bitter orange extract), capsaicin, forskolin (Coleus root extract), and chlorogenic acid (green coffee bean extract). Green tea is an example of a thermogenic with the potential to produce mild but clinically insignificant undesirable stimulatory effects. The use of the aforementioned thermogenic agents in combination with other extracts such as those derived from Salacia reticulata, Sesamum indicum, Lagerstroemia speciosa, Cissus quadrangularis, and Moringa olifera, as well as the use of the carotenoids as lutein and fucoxanthin, and flavonoids as naringin and hesperidin can further facilitate energy metabolism and weight management as well as sports performance without adverse side effects. © 2016 The Authors Phytotherapy Research published by John Wiley & Sons Ltd. PMID:26856274

  5. Influence of the Drilling Mud Formulation Process on the Bacterial Communities in Thermogenic Natural Gas Wells of the Barnett Shale▿†

    OpenAIRE

    Struchtemeyer, Christopher G.; Davis, James P.; Elshahed, Mostafa S.

    2011-01-01

    The Barnett Shale in north central Texas contains natural gas generated by high temperatures (120 to 150°C) during the Mississippian Period (300 to 350 million years ago). In spite of the thermogenic origin of this gas, biogenic sulfide production and microbiologically induced corrosion have been observed at several natural gas wells in this formation. It was hypothesized that microorganisms in drilling muds were responsible for these deleterious effects. Here we collected drilling water and ...

  6. Mineralocorticoid receptor antagonism induces browning of white adipose tissue through impairment of autophagy and prevents adipocyte dysfunction in high-fat-diet-fed mice.

    Science.gov (United States)

    Armani, Andrea; Cinti, Francesca; Marzolla, Vincenzo; Morgan, James; Cranston, Greg A; Antelmi, Antonella; Carpinelli, Giulia; Canese, Rossella; Pagotto, Uberto; Quarta, Carmelo; Malorni, Walter; Matarrese, Paola; Marconi, Matteo; Fabbri, Andrea; Rosano, Giuseppe; Cinti, Saverio; Young, Morag J; Caprio, Massimiliano

    2014-08-01

    The mineralocorticoid receptor (MR) controls adipocyte function, but its role in the conversion of white adipose tissue (WAT) into thermogenic fat has not been elucidated. We investigated responses to the MR antagonists spironolactone (spiro; 20 mg/kg/d) and drospirenone (DRSP; 6 mg/kg/d) in C57BL/6 mice fed a high-fat (HF) diet for 90 d. DRSP and spiro curbed HF diet-induced impairment in glucose tolerance, and prevented body weight gain and white fat expansion. Notably, either MR antagonist induced up-regulation of brown adipocyte-specific transcripts and markedly increased protein levels of uncoupling protein 1 (UCP1) in visceral and inguinal fat depots when compared with the HF diet group. Positron emission tomography and magnetic resonance spectroscopy confirmed acquisition of brown fat features in WAT. Interestingly, MR antagonists markedly reduced the autophagic rate both in murine preadipocytes in vitro (10(-5) M) and in WAT depots in vivo, with a concomitant increase in UCP1 protein expression. Moreover, the autophagy repressor bafilomycin A1 (10(-8) M) mimicked the effect of MR antagonists, increasing UCP1 protein expression in primary preadipocytes. Hence, we showed that adipocyte MR regulates brown remodeling of WAT through a modulation of autophagy. These results provide a rationale for the use of MR antagonists to prevent the adverse metabolic consequences of adipocyte dysfunction. © FASEB.

  7. Feeding Obese Diabetic Mice a Genistein Diet Induces Thermogenic and Metabolic Change.

    Science.gov (United States)

    Rockwood, Schuyler; Broderick, Tom L; Al-Nakkash, Layla

    2017-12-20

    Obesity is associated with elevated plasma levels of glucocorticoids and reduced levels of thyroid hormones, both known to effect food intake and energy expenditure. Furthermore, tissue specific glucocorticoid metabolism is altered in obesity, increasing insulin resistance and cardiometabolic risk. The goal of this study was to examine whether these metabolic disturbances can be prevented with the isoflavone genistein in the ob/ob mouse, a model that resembles the phenotype in human obesity. Male ob/ob mice, aged 5 weeks, were fed either a genistein-rich diet (600 mg/kg) or a genistein-free diet for 4 weeks. ob/ob mice weighed 70% more than lean controls. While there was no effect of genistein on body weight, food consumption during weeks 3 and 4 was significantly increased in genistein-fed mice. This was associated with increases in body temperature and plasma levels of triiodothyronine (T3), suggesting a thermogenic effect. The hypercorticosteronism observed in the ob/ob mouse was reduced with genistein treatment. This effect was accompanied by a decrease in protein expression of renal 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) without changes in hepatic 11β-HSD1. Our results suggest that a diet containing genistein can have beneficial effects on energy expenditure, T3 production, and corticosterone status in the ob/ob mouse model of obesity.

  8. High Grade Myofibroblastic Sarcoma of Paratesticular Soft Tissues

    Directory of Open Access Journals (Sweden)

    Ioannis Anastasiou

    2014-01-01

    Full Text Available Tumors of the paratesticular region most often arise from the soft tissue surrounding the spermatic cord and the epididymis or from the soft tissue (dartos muscle of the scrotal wall. Paratesticular tumors, despite their rarity, present a high incidence of malignancy (30%, and the therapeutic approach of choice is surgical resection with negative margin. The grade, the histology type, the presence of metastases during the diagnosis, the size of the tumor, the age of the patients, and the surgical margins are all important prognostic factors. We present a case report of a 86-year-old patient with a high grade paratesticular and scrotum sarcoma of soft tissues which was presented as a hard painful mass of the scrotum. The patient was subjected to high ligation of the spermatic cord and received no further treatment and 6 months after the operation no local or systematic recurrence was observed.

  9. High-resolution analysis of the mechanical behavior of tissue

    Science.gov (United States)

    Hudnut, Alexa W.; Armani, Andrea M.

    2017-06-01

    The mechanical behavior and properties of biomaterials, such as tissue, have been directly and indirectly connected to numerous malignant physiological states. For example, an increase in the Young's Modulus of tissue can be indicative of cancer. Due to the heterogeneity of biomaterials, it is extremely important to perform these measurements using whole or unprocessed tissue because the tissue matrix contains important information about the intercellular interactions and the structure. Thus, developing high-resolution approaches that can accurately measure the elasticity of unprocessed tissue samples is of great interest. Unfortunately, conventional elastography methods such as atomic force microscopy, compression testing, and ultrasound elastography either require sample processing or have poor resolution. In the present work, we demonstrate the characterization of unprocessed salmon muscle using an optical polarimetric elastography system. We compare the results of compression testing within different samples of salmon skeletal muscle with different numbers of collagen membranes to characterize differences in heterogeneity. Using the intrinsic collagen membranes as markers, we determine the resolution of the system when testing biomaterials. The device reproducibly measures the stiffness of the tissues at variable strains. By analyzing the amount of energy lost by the sample during compression, collagen membranes that are 500 μm in size are detected.

  10. A high definition Mueller polarimetric endoscope for tissue characterisation

    Science.gov (United States)

    Qi, Ji; Elson, Daniel S.

    2016-05-01

    The contrast mechanism of medical endoscopy is mainly based on metrics of optical intensity and wavelength. As another fundamental property of light, polarization can not only reveal tissue scattering and absorption information from a different perspective, but can also provide insight into directional tissue birefringence properties to monitor pathological changes in collagen and elastin. Here we demonstrate a low cost wide field high definition Mueller polarimetric endoscope with minimal alterations to a rigid endoscope. We show that this novel endoscopic imaging modality is able to provide a number of image contrast mechanisms besides traditional unpolarized radiation intensity, including linear depolarization, circular depolarization, cross-polarization, directional birefringence and dichroism. This enhances tissue features of interest, and additionally reveals tissue micro-structure and composition, which is of central importance for tissue diagnosis and image guidance for surgery. The potential applications of the Mueller polarimetric endoscope include wide field early epithelial cancer diagnosis, surgical margin detection and energy-based tissue fusion monitoring, and could further benefit a wide range of endoscopic investigations through intra-operative guidance.

  11. Stream measurements locate thermogenic methane fluxes in groundwater discharge in an area of shale-gas development.

    Science.gov (United States)

    Heilweil, Victor M; Grieve, Paul L; Hynek, Scott A; Brantley, Susan L; Solomon, D Kip; Risser, Dennis W

    2015-04-07

    The environmental impacts of shale-gas development on water resources, including methane migration to shallow groundwater, have been difficult to assess. Monitoring around gas wells is generally limited to domestic water-supply wells, which often are not situated along predominant groundwater flow paths. A new concept is tested here: combining stream hydrocarbon and noble-gas measurements with reach mass-balance modeling to estimate thermogenic methane concentrations and fluxes in groundwater discharging to streams and to constrain methane sources. In the Marcellus Formation shale-gas play of northern Pennsylvania (U.S.A.), we sampled methane in 15 streams as a reconnaissance tool to locate methane-laden groundwater discharge: concentrations up to 69 μg L(-1) were observed, with four streams ≥ 5 μg L(-1). Geochemical analyses of water from one stream with high methane (Sugar Run, Lycoming County) were consistent with Middle Devonian gases. After sampling was completed, we learned of a state regulator investigation of stray-gas migration from a nearby Marcellus Formation gas well. Modeling indicates a groundwater thermogenic methane flux of about 0.5 kg d(-1) discharging into Sugar Run, possibly from this fugitive gas source. Since flow paths often coalesce into gaining streams, stream methane monitoring provides the first watershed-scale method to assess groundwater contamination from shale-gas development.

  12. Basal metabolic rate, maximum thermogenic capacity and aerobic scope in rodents: interaction between environmental temperature and torpor use.

    Science.gov (United States)

    Careau, Vincent

    2013-04-23

    When torpid animals arouse and warm up to restore normal body temperature (T(b)), they produce heat at levels that can reach up to 10 times basal metabolic rate (BMR), close to the cold-induced summit metabolism (VO(2)-sum). Because torpor is an adaptation aimed at conserving energy over periods of low ambient temperature (T(a)) and food availability, selective forces that have led to the evolution of torpor may have simultaneously favoured high thermogenic capacity (i.e. VO(2)-sum) relative to the maintenance costs (i.e. BMR), hence a higher factorial aerobic scope (FAS; the ratio of VO(2)-sum to BMR). My objective was to test this adaptive hypothesis using a phylogenetically informed comparative approach with data on BMR and VO(2)-sum in rodents. I found a strong negative correlation between FAS and the average of the daily minimum T(a) (T(min)) in species using torpor, which was due to differential effects of T(a) on BMR (but not VO(2)-sum) in species that use torpor compared with species that do not. In addition, FAS was negatively correlated with the lowest torpid T(b) in a subset of nine species. These results suggest that in species using torpor, selective forces may have acted to maximize the efficiency of thermogenic capacity (VO(2)-sum) relative to maintenance costs (BMR), resulting in an increasing FAS with decreasing T(a).

  13. Organ and tissue donation: what do high school students know?

    OpenAIRE

    Marcela Cristina de Lemos; Fernanda Myashiro Kian; Alessandra Santos Silva; Tatiana Issida Fujinami; Bartira de Aguiar Roza; Renata Fabiana de Leite; Janine Schirmer

    2007-01-01

    Objectives: To know the opinion of senior high school students in publicand private schools on the process of donating and transplanting organsand tissues, and their desire to be donors. Methods: A descriptive crosssectionalstudy, conducted from 2004 to 2005, on the opinion/knowledgeof senior high school students in public and private schools in the VilaMariana region of the city of São Paulo, on the process of organ and tissuedonation and transplantation. The convenience sample was made up o...

  14. An Updated Protocol for High Throughput Plant Tissue Sectioning

    Directory of Open Access Journals (Sweden)

    Jonathan A. Atkinson

    2017-10-01

    Full Text Available Quantification of the tissue and cellular structure of plant material is essential for the study of a variety of plant sciences applications. Currently, many methods for sectioning plant material are either low throughput or involve free-hand sectioning which requires a significant amount of practice. Here, we present an updated method to provide rapid and high-quality cross sections, primarily of root tissue but which can also be readily applied to other tissues such as leaves or stems. To increase the throughput of traditional agarose embedding and sectioning, custom designed 3D printed molds were utilized to embed 5–15 roots in a block for sectioning in a single cut. A single fluorescent stain in combination with laser scanning confocal microscopy was used to obtain high quality images of thick sections. The provided CAD files allow production of the embedding molds described here from a number of online 3D printing services. Although originally developed for roots, this method provides rapid, high quality cross sections of many plant tissue types, making it suitable for use in forward genetic screens for differences in specific cell structures or developmental changes. To demonstrate the utility of the technique, the two parent lines of the wheat (Triticum aestivum Chinese Spring × Paragon doubled haploid mapping population were phenotyped for root anatomical differences. Significant differences in adventitious cross section area, stele area, xylem, phloem, metaxylem, and cortical cell file count were found.

  15. The thermogenic effect of nesfatin-1 requires recruitment of the melanocortin system.

    Science.gov (United States)

    Dore, Riccardo; Levata, Luka; Gachkar, Sogol; Jöhren, Olaf; Mittag, Jens; Lehnert, Hendrik; Schulz, Carla

    2017-11-01

    Nesfatin-1 is a bioactive polypeptide expressed both in the brain and peripheral tissues and involved in the control of energy balance by reducing food intake. Central administration of nesfatin-1 significantly increases energy expenditure, as demonstrated by a higher dry heat loss; yet, the mechanisms underlying the thermogenic effect of central nesfatin-1 remain unknown. Therefore, in this study, we sought to investigate whether the increase in energy expenditure induced by nesfatin-1 is mediated by the central melanocortin pathway, which was previously reported to mediate central nesfatin-1´s effects on feeding and numerous other physiological functions. With the application of direct calorimetry, we found that intracerebroventricular nesfatin-1 (25 pmol) treatment increased dry heat loss and that this effect was fully blocked by simultaneous administration of an equimolar dose of the melanocortin 3/4 receptor antagonist, SHU9119. Interestingly, the nesfatin-1-induced increase in dry heat loss was positively correlated with body weight loss. In addition, as assessed with thermal imaging, intracerebroventricular nesfatin-1 (100 pmol) increased interscapular brown adipose tissue (iBAT) as well as tail temperature, suggesting increased heat production in the iBAT and heat dissipation over the tail surface. Finally, nesfatin-1 upregulated pro-opiomelanocortin and melanocortin 3 receptor mRNA expression in the hypothalamus, accompanied by a significant increase in iodothyronine deiodinase 2 and by a nonsignificant increase in uncoupling protein 1 and peroxisome proliferator-activated receptor gamma coactivator-1 alpha mRNA in the iBAT. Overall, we clearly demonstrate that nesfatin-1 requires the activation of the central melanocortin system to increase iBAT thermogenesis and, in turn, overall energy expenditure. © 2017 Society for Endocrinology.

  16. Shorebirds' seasonal adjustments in thermogenic capacity are reflected by changes in body mass: how preprogrammed and instantaneous acclimation work together.

    Science.gov (United States)

    Vézina, François; Dekinga, Anne; Piersma, Theunis

    2011-09-01

    metabolically active tissues. Just as cold-exposed birds, red knots in the variable treatment increased body mass in winter, thereby improving cold endurance. During summer, however, they maintained a lower body mass and thermogenic capacity compared to cold-exposed birds, similar to individuals kept at thermoneutrality. We conclude that red knots acclimate to seasonal variations in ambient temperature by modulating body mass, combining a preprogrammed increase in mass during winter with a capacity for fine-tuning body mass and thermogenic capacity to temperature variations.

  17. Characterization of ryanodine receptor and Ca2+-ATPase isoforms in the thermogenic heater organ of blue marlin (Makaira nigricans).

    Science.gov (United States)

    Morrissette, Jeffery M; Franck, Jens P G; Block, Barbara A

    2003-03-01

    A thermogenic organ is found beneath the brain of billfishes (Istiophoridae), swordfish (Xiphiidae) and the butterfly mackerel (Scombridae). The heater organ has been shown to warm the brain and eyes up to 14 degrees C above ambient water temperature. Heater cells are derived from extraocular muscle fibers and express a modified muscle phenotype with an extensive transverse-tubule (T-tubule) network and sarcoplasmic reticulum (SR) enriched in Ca(2+)-ATPase (SERCA) pumps and ryanodine receptors (RyRs). Heater cells have a high mitochondria content but have lost most of the contractile myofilaments. Thermogenesis has been hypothesized to be associated with release and reuptake of Ca(2+). In this study, Ca(2+) fluxes in heater SR vesicles derived from blue marlin (Makaira nigricans) were measured using fura-2 fluorescence. Upon the addition of MgATP, heater SR vesicles rapidly sequestered Ca(2+). Uptake of Ca(2+) was thapsigargin sensitive, and maximum loading ranged between 0.8 micro mol Ca(2+) mg(-1) protein and 1.0 micro mol Ca(2+) mg(-1) protein. Upon the addition of 10 mmol l(-1) caffeine or 350 micro mol l(-1) ryanodine, heater SR vesicles released only a small fraction of the loaded Ca(2+). However, ryanodine could elicit a much larger Ca(2+) release event when the activity of the SERCA pumps was reduced. RNase protection assays revealed that heater tissue expresses an RyR isoform that is also expressed in fish slow-twitch skeletal muscle but is distinct from the RyR expressed in fish fast-twitch skeletal muscle. The heater and slow-twitch muscle RyR isoform has unique physiological properties. In the presence of adenine nucleotides, this RyR remains open even though cytoplasmic Ca(2+) is elevated, a condition that normally closes RyRs. The fast Ca(2+) sequestration by the heater SR, coupled with a physiologically unique RyR, is hypothesized to promote Ca(2+) cycling, ATP turnover and heat generation. A branch of the oculomotor nerve innervates heater organs

  18. High-Speed Coherent Raman Fingerprint Imaging of Biological Tissues

    CERN Document Server

    Camp, Charles H; Heddleston, John M; Hartshorn, Christopher M; Walker, Angela R Hight; Rich, Jeremy N; Lathia, Justin D; Cicerone, Marcus T

    2014-01-01

    We have developed a coherent Raman imaging platform using broadband coherent anti-Stokes Raman scattering (BCARS) that provides an unprecedented combination of speed, sensitivity, and spectral breadth. The system utilizes a unique configuration of laser sources that probes the Raman spectrum over 3,000 cm$^{-1}$ and generates an especially strong response in the typically weak Raman "fingerprint" region through heterodyne amplification of the anti-Stokes photons with a large nonresonant background (NRB) while maintaining high spectral resolution of $<$ 13 cm$^{-1}$. For histology and pathology, this system shows promise in highlighting major tissue components in a non-destructive, label-free manner. We demonstrate high-speed chemical imaging in two- and three-dimensional views of healthy murine liver and pancreas tissues and interfaces between xenograft brain tumors and the surrounding healthy brain matter.

  19. Maternal dietary fat affects milk fatty acid profile and impacts on weight gain and thermogenic capacity of suckling rats.

    Science.gov (United States)

    Priego, Teresa; Sánchez, Juana; García, Ana Paula; Palou, Andreu; Picó, Catalina

    2013-05-01

    We aimed to assess the effects of maternal supplementation with the main fat sources used in the human Western diet (olive oil, butter, margarine) on milk FA composition and on plasma FA profile of offspring, and to determine whether it may influence body-weight-gain (BWG) and adiposity of offspring during the suckling period. Wistar rats were supplemented with the different fat sources from day 14 of gestation and throughout lactation. Olive oil-supplemented dams showed the highest proportion of oleic-acid in milk, with no changes in plasma. Their offspring also showed the highest proportion of this FA in plasma, lower BWG during the suckling period, and higher levels of UCP1 in brown adipose tissue (BAT) at weaning. Margarine-supplemented dams showed the highest percentage of PUFA in milk, and a similar tendency was found in plasma of their offspring. Butter-supplemented dams displayed higher proportion of saturated FA (SFA) in milk compared to other fat-supplemented dams, but lower than controls. Control offspring also showed higher proportion of SFA in plasma and greater BWG during the suckling period than fat-supplemented groups. Significant correlations were found between the relative content of some milk FA and BWG of offspring, in particular, oleic-acid levels correlated negatively with BWG and positively with UCP1 levels. These results show that maternal dietary source of fat affects milk FA composition and circulating FA profile, as could be expected, but also BWG and thermogenic capacity of offspring during the suckling period. An effect of oleic-acid stimulating BAT thermogenic capacity of suckling pups is proposed.

  20. Loss of perivascular adipose tissue on peroxisome proliferator-activated receptor-γ deletion in smooth muscle cells impairs intravascular thermoregulation and enhances atherosclerosis.

    Science.gov (United States)

    Chang, Lin; Villacorta, Luis; Li, Rongxia; Hamblin, Milton; Xu, Wei; Dou, Chunyan; Zhang, Jifeng; Wu, Jiarui; Zeng, Rong; Chen, Y Eugene

    2012-08-28

    Perivascular adipose tissue (PVAT) surrounds most vessels and shares common features with brown adipose tissue (BAT). Although adaptive thermogenesis in BAT increases energy expenditure and is beneficial for metabolic diseases, little is known about the role of PVAT in vascular diseases such as atherosclerosis. We hypothesize that the thermogenic function of PVAT regulates intravascular temperature and reduces atherosclerosis. PVAT shares similar structural and proteomics with BAT. We demonstrated that PVAT has thermogenic properties similar to BAT in response to cold stimuli in vivo. Proteomics analysis of the PVAT from mice housed in a cold environment identified differential expression in proteins highly related to cellular metabolic processes. In a mouse model deficient in peroxisome proliferator-activated receptor-γ in smooth muscle cells (SMPG KO mice), we uncovered a complete absence of PVAT surrounding the vasculature, likely caused by peroxisome proliferator-activated receptor-γ deletion in the perivascular adipocyte precursor cells as well. Lack of PVAT, which results in loss of its thermogenic activity, impaired vascular homeostasis, which caused temperature loss and endothelial dysfunction. We further showed that cold exposure inhibits atherosclerosis and improves endothelial function in mice with intact PVAT but not in SMPG KO mice as a result of impaired lipid clearance. Proinflammatory cytokine expression in PVAT is not altered on exposure to cold. Finally, prostacyclin released from PVAT contributes to the vascular protection against endothelial dysfunction. PVAT is a vasoactive organ with functional characteristics similar to BAT and is essential for intravascular thermoregulation of cold acclimation. This thermogenic capacity of PVAT plays an important protective role in the pathogenesis of atherosclerosis.

  1. A High Rate Tension Device for Characterizing Brain Tissue

    CERN Document Server

    Rashid, Badar; Gilchrist, Michael; 10.1177/1754337112436900

    2013-01-01

    The mechanical characterization of brain tissue at high loading velocities is vital for understanding and modeling Traumatic Brain Injury (TBI). The most severe form of TBI is diffuse axonal injury (DAI) which involves damage to individual nerve cells (neurons). DAI in animals and humans occurs at strains > 10% and strain rates > 10/s. The mechanical properties of brain tissues at these strains and strain rates are of particular significance, as they can be used in finite element human head models to accurately predict brain injuries under different impact conditions. Existing conventional tensile testing machines can only achieve maximum loading velocities of 500 mm/min, whereas the Kolsky bar apparatus is more suitable for strain rates > 100/s. In this study, a custom-designed high rate tension device is developed and calibrated to estimate the mechanical properties of brain tissue in tension at strain rates < 90/s, while maintaining a uniform velocity. The range of strain can also be extended to 100% de...

  2. A high throughput mechanical screening device for cartilage tissue engineering.

    Science.gov (United States)

    Mohanraj, Bhavana; Hou, Chieh; Meloni, Gregory R; Cosgrove, Brian D; Dodge, George R; Mauck, Robert L

    2014-06-27

    Articular cartilage enables efficient and near-frictionless load transmission, but suffers from poor inherent healing capacity. As such, cartilage tissue engineering strategies have focused on mimicking both compositional and mechanical properties of native tissue in order to provide effective repair materials for the treatment of damaged or degenerated joint surfaces. However, given the large number design parameters available (e.g. cell sources, scaffold designs, and growth factors), it is difficult to conduct combinatorial experiments of engineered cartilage. This is particularly exacerbated when mechanical properties are a primary outcome, given the long time required for testing of individual samples. High throughput screening is utilized widely in the pharmaceutical industry to rapidly and cost-effectively assess the effects of thousands of compounds for therapeutic discovery. Here we adapted this approach to develop a high throughput mechanical screening (HTMS) system capable of measuring the mechanical properties of up to 48 materials simultaneously. The HTMS device was validated by testing various biomaterials and engineered cartilage constructs and by comparing the HTMS results to those derived from conventional single sample compression tests. Further evaluation showed that the HTMS system was capable of distinguishing and identifying 'hits', or factors that influence the degree of tissue maturation. Future iterations of this device will focus on reducing data variability, increasing force sensitivity and range, as well as scaling-up to even larger (96-well) formats. This HTMS device provides a novel tool for cartilage tissue engineering, freeing experimental design from the limitations of mechanical testing throughput. © 2013 Published by Elsevier Ltd.

  3. The suppression of ghrelin signaling mitigates age-associated thermogenic impairment

    Science.gov (United States)

    Aging is associated with severe thermogenic impairment, which contributes to obesity and diabetes in aging. We previously reported that ablation of the ghrelin receptor, growth hormone secretagogue receptor (GHS-R), attenuates age-associated obesity and insulin resistance. Ghrelin and obestatin are ...

  4. Breast tissue characterization with high-frequency scanning acoustic microscopy

    Science.gov (United States)

    Kumon, R. E.; Bruno, I.; Heartwell, B.; Maeva, E.

    2004-05-01

    We have performed imaging of breast tissue using scanning acoustic microscopy (SAM) in the range of 25-50 MHz with the goal of accurately and rapidly determining the structure and composition throughout the volume of the samples. In contrast to traditional histological slides, SAM images can be obtained without special preparation, sometimes even without sectioning, but with sufficiently high spatial resolution to give information comparable to surface optical images. As a result, the use of high-frequency SAM at the time of breast lumpectomy to identify disease-free margins has the potential to reduce reoperative rates, patient anxiety, and local recurrence. However, only limited work has been performed to characterize breast tissue in the frequency range above clinical ultrasound devices. The samples are 4-cm2-thick sections (2-3 mm) taken from mastectomies and preserved in formalin. They are placed between two plates and immersed in water during imaging. Attenuation images are acquired by focusing the acoustic beam at the top and bottom of the samples, although better results were obtained for bottom focusing. For purposes of comparison and identification of histological features, acoustical images will be presented along with optical images obtained from the same samples. [Work supported by CIHR.

  5. Resistance to the anorexic and thermogenic effects of centrally administrated leptin in obese aged rats.

    Science.gov (United States)

    Shek, E W; Scarpace, P J

    2000-08-25

    The aim of the present study was to determine whether the anorexic and thermogenic effects of leptin were attenuated in overweight aged rats following intracerebroventricular (i.c.v.) injection of murine leptin. Male F344/BN rats of two ages (6 months: young (n=20) and 24 months: old (n=18)) were divided into three groups (control, pair-fed and leptin) and were treated with either vehicle (artificial cerebrospinal fluid) or leptin (15.6 microgram/day) for 3 days. There was an age-related increase in basal food intake (20+/-2%), serum leptin levels (363+/-106%) and leptin (OB) mRNA (72+/-16%) in perirenal white adipose tissue (PWAT). In contrast, basal expression of hypothalamic NPY mRNA and brown adipose tissue (BAT) uncoupling protein 1 (UCP1) mRNA was reduced significantly (-35+/-4% and -51+/-5%, respectively) with age. I.c.v. leptin treatment had a significantly greater effect in reducing food intake (-42+/-5% vs. -23+/-4%), serum leptin levels (-55+/-7% vs. 10+/-2%) and PWAT OB mRNA (-46+/-2% vs. 10+/-5%) in young than in old rats. Similarly, central leptin treatment also had a greater effect in suppressing hypothalamic NPY mRNA expression in young (-23+/-4%) than in old (-8+/-4%) rats compared with their age-matched pair-fed treated rats. The stimulatory effect of i.c.v. leptin treatment on BAT UCP1 mRNA expression was also significantly greater in young rats (45+/-8%) than in old rats (10+/-6%) compared with age-matched pair-fed rats. Our previous report indicated that these overweight aged rats were resistant to peripheral administered leptin. The present data extend those findings and demonstrate that the impaired anorexic and metabolic effects of leptin are centrally mediated. This leptin resistance may be due to either the elevated obesity and serum leptin with age or due to age itself or both. The development of leptin resistance with age may contribute to the hyperphagia, hyperleptinemia and impaired energy balance with age.

  6. Organ and tissue donation: what do high school students know?

    Directory of Open Access Journals (Sweden)

    Marcela Cristina de Lemos

    2007-09-01

    Full Text Available Objectives: To know the opinion of senior high school students in publicand private schools on the process of donating and transplanting organsand tissues, and their desire to be donors. Methods: A descriptive crosssectionalstudy, conducted from 2004 to 2005, on the opinion/knowledgeof senior high school students in public and private schools in the VilaMariana region of the city of São Paulo, on the process of organ and tissuedonation and transplantation. The convenience sample was made up of140 (81% students from two private schools and 167 (51% studentsfrom a public school. The project was approved by the Research EthicsCommittee of the UNIFESP. Results: Data showed that 163 (53.1%students believe that donation is by presumed consent and 147 (47.9%that consider that it occurs by informed consent. Of the public schoolstudents, 120 (71.9% believe that transplants are public and free ofcharge in Brazil versus 94 (67.1% of the students from private schools.Students know that donations may be made by living or dead donors(121 - 86.4% private schools versus 113 – 67.7% public school. Wehighlight that 22 (15.7% of the private school students and 16 (9.6%of those from the public school believe that the commerce of organs isallowed in Brazil. As to intentions of being a donor, 108 (77.1% of theprivate school students declared themselves organ and tissue donorsversus 106 (63.5% from the public school, and 63 (59.4% from thepublic versus 61 (56.5% from the private schools have already informedtheir families. Conclusion: There was no difference in knowledge andopinion among the students from the public and private schools as toaspects regarding donation and transplantation.

  7. Brown adipose tissue in the treatment of obesity and diabetes: Are we hot enough?

    Science.gov (United States)

    Tan, Chong Yew; Ishikawa, Ko; Virtue, Samuel; Vidal-Puig, Antonio

    2011-10-07

    The identification of functional brown adipose tissue in human adults has intensified interest in exploiting thermogenic energy expenditure for the purpose of weight management. However, food intake and energy expenditure are tightly regulated and it is generally accepted that variation in one component results in compensatory changes in the other. In the context of weight loss, additional biological adaptations occur in an attempt to further limit weight loss. In the present review, we discuss the relationship between increasing energy expenditure and body weight in humans, including the effects of cold exposure. The data raise the possibility that some processes, particularly those involved in thermogenesis, induce less compensatory food intake for a given magnitude of additional energy expenditure, a state we term the 'thermogenic disconnect'. Although cold exposure increases thermogenesis and can putatively be exploited to induce weight loss, there are multiple adaptive responses to cold, of which many actually reduce energy expenditure. In order to optimally exploit either cold itself or agents that mimic cold for thermogenic energy expenditure, these non-thermogenic cold responses must be considered. Finally, the relative contribution of brown adipose tissue vs other thermogenic processes in humans remains to be defined. However, overall the data suggest that activation of cold-induced thermogenic processes are promising targets for interventions to treat obesity and its secondary metabolic complications. (J Diabetes Invest, doi:10.1111/j.2040-1124.2011.00158.x, 2011).

  8. Anatomic composition of plant tissues of highly metamorphosed coals

    Energy Technology Data Exchange (ETDEWEB)

    Kizil' shtein, L.Ya.; Shpitsgluz, A.L.

    1985-09-01

    Method is described to improve microscopic study of highly metamorphosed coals (anthracite). Study of such coals with aid of reflected polarized light is enhanced by means of ionic etching of surface of slides that enables observation not only of structures of basic microcomponents but also of finest structural details of individual cells by reflected non-polarized light. Figures illustrate results of studying many samples by ionic etching (bombing a polished surface in a vacuum with ions and pulverizing material of microcomponents to reveal heterogeneity of crystal chemistry of surface) which reveals great variety of structures of plant tissues and their component cells. Pictures of 35 slides depict gelified coal-forming plants of Donbass and central Ural coal fields; fusainized coal-forming plants of Donetsk, Gorlovsk and Tungus basins; organs of Donbass plants; structure of cells and organs of plants of Donbass. Method of ionic etching opens new perspectives for studying anatomy and histology in area of classical paleobotany by making available a large number of samples of plant material and components of highly metamorphosed coals compared with the rare samples obtained by using the polarized light method. 14 references.

  9. Thermogenic side effects to migratory predisposition in shorebirds

    NARCIS (Netherlands)

    Vezina, Francois; Jalvingh, Kirsten M.; Dekinga, Anne; Piersma, Theunis

    In the calidrine sandpiper red knot (Calidris canutus), the weeks preceding takeoff for long-distance migration are characterized by a rapid increase in body mass, largely made up of fat but also including a significant proportion of lean tissue. Before takeoff, the pectoral muscles are known to

  10. Leptin-induced leptin resistance reveals separate roles for the anorexic and thermogenic responses in weight maintenance.

    Science.gov (United States)

    Scarpace, Philip J; Matheny, Michael; Zhang, Yi; Shek, Eugene W; Prima, Victor; Zolotukhin, Sergei; Tümer, Nihal

    2002-08-01

    The purpose of this study was to determine whether leptin induces leptin resistance by examining the temporal attenuation of the anorexic and energy expenditure responses to leptin. We administered recombinant adeno-associated virus encoding rat leptin cDNA or control viral vector into mildly obese rats for 138 d and compared these results with those from pair-fed rats. We measured food consumption, body weight, oxygen consumption, leptin signal transduction, and brown adipose tissue uncoupling protein 1. The anorexic response attenuated by d 25, whereas the increase in energy expenditure persisted for 83 d before attenuating. Despite attenuation of physiological responses, phosphorylated signal transducer and activator of transcription-3 remained elevated for the duration of the study. The temporal differential attenuation of the anorexic and thermogenic responses allowed us to determine the relative contributions of each response to weight maintenance. The anorexic response predominantly mediated the initial loss of body weight, but only the energy expenditure response was necessary to maintain the reduced weight. This study provides evidence that leptin induces leptin resistance. The leptin resistance was associated with persistent elevation in hypothalamic phosphorylated signal transducer and activator of transcription-3 and was characterized by a rapid attenuation of the anorexic response and slower onset for the attenuation of the energy expenditure response. We propose that both elevated leptin and obesity may be necessary for the development of leptin resistance.

  11. Amplification of thermostable lipase genes fragment from thermogenic phase of domestic waste composting process

    Science.gov (United States)

    Nurhasanah, Nurbaiti, Santi; Madayanti, Fida; Akhmaloka

    2015-09-01

    Lipases are lipolytic enzymes, catalyze the hydrolysis of fatty acid ester bonds of triglycerides to produce free fatty acids and glycerol. The enzyme is widely used in various fields of biotechnological industry. Hence, lipases with unique properties (e.g.thermostable lipase) are still being explored by variation methods. One of the strategy is by using metagenomic approach to amplify the gene directly from environmental sample. This research was focused on amplification of lipase gene fragment directly from the thermogenic phase of domestic waste composting in aerated trenches. We used domestic waste compost from waste treatment at SABUGA, ITB for the sample. Total chromosomal DNA were directly extracted from several stages at thermogenic phase of compost. The DNA was then directly used as a template for amplification of thermostable lipase gene fragments using a set of internal primers namely Flip-1a and Rlip-1a that has been affixed with a GC clamp in reverse primer. The results showed that the primers amplified the gene from four stages of thermogenic phase with the size of lipase gene fragment of approximately 570 base pairs (bp). These results were further used for Denaturing Gradient Gel Electrophoresis (DGGE) analysis to determine diversity of thermostable lipase gene fragments.

  12. Towards high throughput tissue engineering: development of chitosan-calcium phosphate scaffolds for engineering bone tissue from embryonic stem cells

    OpenAIRE

    Ko, Junghyuk; Kolehmainen, Kathleen; Ahmed, Farid; Jun, Martin BG; Willerth, Stephanie M.

    2011-01-01

    Tissue engineering strategies have shown promise for the repair of damaged organs, including bone. One of the major challenges associated with tissue engineering is how to scale up such processes for high throughput manufacturing of biomaterial scaffolds used to support stem cell culture. Generation of certain types of 3D biomaterial scaffolds, including chitosan-calcium phosphate blends, involves a slow fabrication process followed by a lengthy required freeze drying step. This work investig...

  13. High-Density Spot Seeding for Tissue Model Formation

    Science.gov (United States)

    Marquette, Michele L. (Inventor); Sognier, Marguerite A. (Inventor)

    2016-01-01

    A model of tissue is produced by steps comprising seeding cells at a selected concentration on a support to form a cell spot, incubating the cells to allow the cells to partially attach, rinsing the cells to remove any cells that have not partially attached, adding culture medium to enable the cells to proliferate at a periphery of the cell spot and to differentiate toward a center of the cell spot, and further incubating the cells to form the tissue. The cells may be C2C12 cells or other subclones of the C2 cell line, H9c2(2-1) cells, L6 cells, L8 cells, QM7 cells, Sol8 cells, G-7 cells, G-8 cells, other myoblast cells, cells from other tissues, or stem cells. The selected concentration is in a range from about 1 x 10(exp 5) cells/ml to about 1 x 10(exp 6) cells/ml. The tissue formed may be a muscle tissue or other tissue depending on the cells seeded.

  14. Versatile tissue lasers based on high-Q Fabry-Pérot microcavities.

    Science.gov (United States)

    Chen, Yu-Cheng; Chen, Qiushu; Zhang, Tingting; Wang, Wenjie; Fan, Xudong

    2017-01-31

    Biolasers are an emerging technology for next generation biochemical detection and clinical applications. Progress has recently been made to achieve lasing from biomolecules and single living cells. Tissues, which consist of cells embedded in an extracellular matrix, mimic more closely the actual complex biological environment in a living body and therefore are of more practical significance. Here, we developed a highly versatile tissue laser platform, in which tissues stained with fluorophores are sandwiched in a high-Q Fabry-Pérot microcavity. Distinct lasing emissions from muscle and adipose tissues stained respectively with fluorescein isothiocyanate (FITC) and boron-dipyrromethene (BODIPY), and hybrid muscle/adipose tissue with dual staining were achieved with a threshold of only ∼10 μJ mm-2. Additionally, we investigated how the tissue structure/geometry, tissue thickness, and staining dye concentration affect the tissue laser. Lasing emission from FITC conjugates (FITC-phalloidin) that specifically target F-actin in muscle tissues was also realized. It is further found that, despite the large fluorescence spectral overlap between FITC and BODIPY in tissues, their lasing emissions could be clearly distinguished and controlled due to their narrow lasing bands and different lasing thresholds, thus enabling highly multiplexed detection. Our tissue laser platform can be broadly applicable to various types of tissues/diseases. It provides a new tool for a wide range of biological and biomedical applications, such as diagnostics/screening of tissues and identification/monitoring of biological transformations in tissue engineering.

  15. PexRAP Inhibits PRDM16-Mediated Thermogenic Gene Expression

    Directory of Open Access Journals (Sweden)

    Irfan J. Lodhi

    2017-09-01

    Full Text Available How the nuclear receptor PPARγ regulates the development of two functionally distinct types of adipose tissue, brown and white fat, as well as the browning of white fat, remains unclear. Our previous studies suggest that PexRAP, a peroxisomal lipid synthetic enzyme, regulates PPARγ signaling and white adipogenesis. Here, we show that PexRAP is an inhibitor of brown adipocyte gene expression. PexRAP inactivation promoted adipocyte browning, increased energy expenditure, and decreased adiposity. Identification of PexRAP-interacting proteins suggests that PexRAP function extends beyond its role as a lipid synthetic enzyme. Notably, PexRAP interacts with importin-β1, a nuclear import factor, and knockdown of PexRAP in adipocytes reduced the levels of nuclear phospholipids. PexRAP also interacts with PPARγ, as well as PRDM16, a critical transcriptional regulator of thermogenesis, and disrupts the PRDM16-PPARγ complex, providing a potential mechanism for PexRAP-mediated inhibition of adipocyte browning. These results identify PexRAP as an important regulator of adipose tissue remodeling.

  16. Consuming a multi-ingredient thermogenic supplement for 28 days is apparently safe in healthy adults.

    Science.gov (United States)

    Vogel, Roxanne M; Joy, Jordan M; Falcone, Paul H; Mosman, Matt M; Kim, Michael P; Moon, Jordan R

    2015-01-01

    Thermogenic (TRM) supplements are often used by people seeking to decrease body weight. Many TRM supplements are formulated with multiple ingredients purported to increase energy expenditure and maximize fat loss. However, in the past some TRM ingredients have been deemed unsafe and removed from the market. Therefore, it is important to verify the safety of multi-ingredient TRM supplements with chronic consumption. To assess the safety of daily consumption of a multi-ingredient TRM supplement over a 28-day period in healthy adults. Twenty-three recreationally active adults (11M, 12F; 27.1±5.4 years, 171.6±9.6 cm, 76.8±16.1 kg, 26±5 BMI) were randomly assigned either to consume a multi-ingredient TRM supplement (SUP; n=9) or remain unsupplemented (CRL; n=14) for 28 days. Participants maintained their habitual dietary and exercise routines for the duration of the study. Fasting blood samples, resting blood pressure, and heart rate were taken before and after the supplementation period. Samples were analyzed for complete blood counts, comprehensive metabolic, and lipid panels. Significant (p<0.05) group by time interactions were present for diastolic BP, creatinine, estimated glomerular filtration rate (eGFR), chloride, CO2, globulin, albumin:globulin (A/G), and high-density lipoprotein (HDL). Dependent t-tests conducted on significant variables revealed significant (p<0.05) within-group differences in SUP for diastolic BP (+6.2±5.3 mmHG), creatinine (+0.09±0.05 mg/dL), eGFR (-11.2±5.8 mL/min/1.73), globulin (-0.29±0.24 g/dL), A/G (+0.27±0.23), and HDL (-5.0±5.5 mg/dL), and in CRL for CO2 (-1.9±1.5 mmol/L) between time points. Each variable remained within the accepted physiological range. Results of the present study support the clinical safety of a multi-ingredient TRM containing caffeine, green tea extract, and cayenne powder. Although there were statistically significant (p<0.05) intragroup differences in SUP from pre- to postsupplementation for

  17. Consuming a multi-ingredient thermogenic supplement for 28 days is apparently safe in healthy adults

    Directory of Open Access Journals (Sweden)

    Roxanne M. Vogel

    2015-07-01

    Full Text Available Background: Thermogenic (TRM supplements are often used by people seeking to decrease body weight. Many TRM supplements are formulated with multiple ingredients purported to increase energy expenditure and maximize fat loss. However, in the past some TRM ingredients have been deemed unsafe and removed from the market. Therefore, it is important to verify the safety of multi-ingredient TRM supplements with chronic consumption. Objective: To assess the safety of daily consumption of a multi-ingredient TRM supplement over a 28-day period in healthy adults. Design: Twenty-three recreationally active adults (11M, 12F; 27.1±5.4 years, 171.6±9.6 cm, 76.8±16.1 kg, 26±5 BMI were randomly assigned either to consume a multi-ingredient TRM supplement (SUP; n=9 or remain unsupplemented (CRL; n=14 for 28 days. Participants maintained their habitual dietary and exercise routines for the duration of the study. Fasting blood samples, resting blood pressure, and heart rate were taken before and after the supplementation period. Samples were analyzed for complete blood counts, comprehensive metabolic, and lipid panels. Results: Significant (p<0.05 group by time interactions were present for diastolic BP, creatinine, estimated glomerular filtration rate (eGFR, chloride, CO2, globulin, albumin:globulin (A/G, and high-density lipoprotein (HDL. Dependent t-tests conducted on significant variables revealed significant (p<0.05 within-group differences in SUP for diastolic BP (+6.2±5.3 mmHG, creatinine (+0.09±0.05 mg/dL, eGFR (−11.2±5.8 mL/min/1.73, globulin (−0.29±0.24 g/dL, A/G (+0.27±0.23, and HDL (−5.0±5.5 mg/dL, and in CRL for CO2 (−1.9±1.5 mmol/L between time points. Each variable remained within the accepted physiological range. Conclusion: Results of the present study support the clinical safety of a multi-ingredient TRM containing caffeine, green tea extract, and cayenne powder. Although there were statistically significant (p<0.05 intragroup

  18. High contrast soft tissue imaging based on multi-energy x-ray

    Science.gov (United States)

    Oh, Hyun-Hwa; Sung, Young-Hun; Kim, Sung-Su; Kwon, Jae-Hyun; Lee, Seong-Deok; Kim, Chang-Yeong

    2011-03-01

    Breast soft tissues have similar x-ray attenuations to mass tissue. Overlapping breast tissue structure often obscures mass and microcalcification, essential to the early detection of breast cancer. In this paper, we propose new method to generate the high contrast mammogram with distinctive features of a breast cancer by using multiple images with different x-ray energy spectra. On the experiments with mammography simulation and real breast tissues, the proposed method has provided noticeable images with obvious mass structure and microcalifications.

  19. Kynurenic Acid and Gpr35 Regulate Adipose Tissue Energy Homeostasis and Inflammation

    DEFF Research Database (Denmark)

    Agudelo, Leandro Z; Ferreira, Duarte M S; Cervenka, Igor

    2018-01-01

    accumulation. Here we show that kynurenic acid increases energy utilization by activating G protein-coupled receptor Gpr35, which stimulates lipid metabolism, thermogenic, and anti-inflammatory gene expression in adipose tissue. This suppresses weight gain in animals fed a high-fat diet and improves glucose...... tolerance. Kynurenic acid and Gpr35 enhance Pgc-1α1 expression and cellular respiration, and increase the levels of Rgs14 in adipocytes, which leads to enhanced beta-adrenergic receptor signaling. Conversely, genetic deletion of Gpr35 causes progressive weight gain and glucose intolerance, and sensitizes...... to the effects of high-fat diets. Finally, exercise-induced adipose tissue browning is compromised in Gpr35 knockout animals. This work uncovers kynurenine metabolism as a pathway with therapeutic potential to control energy homeostasis....

  20. Positioning systems for high-resolution tissue imaging

    Science.gov (United States)

    Haylock, Thomas M.; Cenko, Andrew T.; Chifman, Lev M.; Christensen, Peter B.; Kazemzadeh, Farnoud; Hajian, Arsen R.; Hendrikse, Jan; Meade, Jeff T.

    2011-03-01

    Tissue handling systems position ex-vivo samples to a required accuracy that depends on the features to be imaged. For example, to resolve cellular structure, micron pixel spacing is needed. 3D tissue scanning at cellular resolution allows for more complete histology to be obtained and more accurate diagnosis to be made. However, accurate positioning of a light beam on the sample is a significant challenge, especially when fine spacing between scan steps is desired or large, inconsistently shaped samples need to be imaged. Optical coherence tomography (OCT) is an application where accurate positioning systems are required to reap the full benefit of the technology. By simultaneously manipulating the light beam position and sample location, a 3D image is reconstructed from a series of depth profiles produced. To automate image acquisition, a fully integrated and synchronised system is necessary. A tissue handling and light delivery system for free-space optical devices is described. Performance characteristics such as resolution, uncertainty, and repeatability are evaluated for novel hardware configurations of OCT. Typical scanning patterns with associated synchronisation requirements are discussed.

  1. Developing High-Frequency Quantitative Ultrasound Techniques to Characterize Three-Dimensional Engineered Tissues

    Science.gov (United States)

    Mercado, Karla Patricia E.

    Tissue engineering holds great promise for the repair or replacement of native tissues and organs. Further advancements in the fabrication of functional engineered tissues are partly dependent on developing new and improved technologies to monitor the properties of engineered tissues volumetrically, quantitatively, noninvasively, and nondestructively over time. Currently, engineered tissues are evaluated during fabrication using histology, biochemical assays, and direct mechanical tests. However, these techniques destroy tissue samples and, therefore, lack the capability for real-time, longitudinal monitoring. The research reported in this thesis developed nondestructive, noninvasive approaches to characterize the structural, biological, and mechanical properties of 3-D engineered tissues using high-frequency quantitative ultrasound and elastography technologies. A quantitative ultrasound technique, using a system-independent parameter known as the integrated backscatter coefficient (IBC), was employed to visualize and quantify structural properties of engineered tissues. Specifically, the IBC was demonstrated to estimate cell concentration and quantitatively detect differences in the microstructure of 3-D collagen hydrogels. Additionally, the feasibility of an ultrasound elastography technique called Single Tracking Location Acoustic Radiation Force Impulse (STL-ARFI) imaging was demonstrated for estimating the shear moduli of 3-D engineered tissues. High-frequency ultrasound techniques can be easily integrated into sterile environments necessary for tissue engineering. Furthermore, these high-frequency quantitative ultrasound techniques can enable noninvasive, volumetric characterization of the structural, biological, and mechanical properties of engineered tissues during fabrication and post-implantation.

  2. Intra-seasonal flexibility in avian metabolic performance highlights the uncoupling of basal metabolic rate and thermogenic capacity.

    Science.gov (United States)

    Petit, Magali; Lewden, Agnès; Vézina, François

    2013-01-01

    Stochastic winter weather events are predicted to increase in occurrence and amplitude at northern latitudes and organisms are expected to cope through phenotypic flexibility. Small avian species wintering in these environments show acclimatization where basal metabolic rate (BMR) and maximal thermogenic capacity (MSUM) are typically elevated. However, little is known on intra-seasonal variation in metabolic performance and on how population trends truly reflect individual flexibility. Here we report intra-seasonal variation in metabolic parameters measured at the population and individual levels in black-capped chickadees (Poecileatricapillus). Results confirmed that population patterns indeed reflect flexibility at the individual level. They showed the expected increase in BMR (6%) and MSUM (34%) in winter relative to summer but also, and most importantly, that these parameters changed differently through time. BMR began its seasonal increase in November, while MSUM had already achieved more than 20% of its inter-seasonal increase by October, and declined to its starting level by March, while MSUM remained high. Although both parameters co-vary on a yearly scale, this mismatch in the timing of variation in winter BMR and MSUM likely reflects different constraints acting on different physiological components and therefore suggests a lack of functional link between these parameters.

  3. Influence of the drilling mud formulation process on the bacterial communities in thermogenic natural gas wells of the Barnett Shale.

    Science.gov (United States)

    Struchtemeyer, Christopher G; Davis, James P; Elshahed, Mostafa S

    2011-07-01

    The Barnett Shale in north central Texas contains natural gas generated by high temperatures (120 to 150°C) during the Mississippian Period (300 to 350 million years ago). In spite of the thermogenic origin of this gas, biogenic sulfide production and microbiologically induced corrosion have been observed at several natural gas wells in this formation. It was hypothesized that microorganisms in drilling muds were responsible for these deleterious effects. Here we collected drilling water and drilling mud samples from seven wells in the Barnett Shale during the drilling process. Using quantitative real-time PCR and microbial enumerations, we show that the addition of mud components to drilling water increased total bacterial numbers, as well as the numbers of culturable aerobic heterotrophs, acid producers, and sulfate reducers. The addition of sterile drilling muds to microcosms that contained drilling water stimulated sulfide production. Pyrosequencing-based phylogenetic surveys of the microbial communities in drilling waters and drilling muds showed a marked transition from typical freshwater communities to less diverse communities dominated by Firmicutes and Gammaproteobacteria. The community shifts observed reflected changes in temperature, pH, oxygen availability, and concentrations of sulfate, sulfonate, and carbon additives associated with the mud formulation process. Finally, several of the phylotypes observed in drilling muds belonged to lineages that were thought to be indigenous to marine and terrestrial fossil fuel formations. Our results suggest a possible alternative exogenous origin of such phylotypes via enrichment and introduction to oil and natural gas reservoirs during the drilling process.

  4. Comparison of the thermogenic and hypophagic effects of sibutramine's metabolite 2 and other monoamine reuptake inhibitors.

    Science.gov (United States)

    Liu, Yong-Ling; Connoley, Ian P; Harrison, Jonathan; Heal, David J; Stock, Michael J

    2002-09-27

    The thermogenic and hypophagic effects of sibutramine's metabolite (metabolite 2), a 5-hydroxytryptamine (5-HT) and noradrenaline reuptake inhibitor, were investigated in rats and compared with duloxetine and bupropion. Metabolite 2 increased colonic temperature for 2.5-4.5 h. Duloxetine was also thermogenic but was less effective than metabolite 2. Bupropion similarly increased colonic temperature and was as efficacious, but less potent, than metabolite 2. At -8 degrees C, metabolite 2, duloxetine and bupropion all decreased response to heat reinforcement and reduced colonic temperature. Metabolite 2 produced a sustained increase in oxygen consumption (VO(2)) at 29 degrees C from 90 to 240 min, whereas duloxetine was far less effective. Bupropion rapidly enhanced VO(2), but its effect was less prolonged than that of metabolite 2. Metabolite 2 markedly reduced 24-h food intake. Duloxetine decreased feeding although its effect was shorter-lived, but bupropion was without effect. Thus, sibutramine's antiobesity action is probably attributable to effects on energy intake and expenditure. Duloxetine shares these properties, but is generally less efficacious. Any potential weight-reducing effect of bupropion is probably due to thermogenesis. Copyright 2002 Elsevier Science B.V.

  5. A Novel Approach to High-Quality Postmortem Tissue Procurement: The GTEx Project

    NARCIS (Netherlands)

    Carithers, Latarsha J.; Ardlie, Kristin; Barcus, Mary; Branton, Philip A.; Britton, Angela; Buia, Stephen A.; Compton, Carolyn C.; Deluca, David S.; Peter-Demchok, Joanne; Gelfand, Ellen T.; Guan, Ping; Korzeniewski, Greg E.; Lockhart, Nicole C.; Rabiner, Chana A.; Rao, Abhi K.; Robinson, Karna L.; Roche, Nancy V.; Sawyer, Sherilyn J.; Segrè, Ayellet V.; Shive, Charles E.; Smith, Anna M.; Sobin, Leslie H.; Undale, Anita H.; Valentino, Kimberly M.; Vaught, Jim; Young, Taylor R.; Moore, Helen M.; Barker, Laura; Basile, Margaret; Battle, Alexis; Boyer, Joy; Bradbury, Debra; Bridge, Jason P.; Brown, Amanda; Burges, Robin; Choi, Christopher; Colantuoni, Deborah; Cox, Nancy; Dermitzakis, Emmanouil T.; Derr, Leslie K.; Dinsmore, Michael J.; Erickson, Kenyon; Fleming, Johnelle; Flutre, Timothée; Foster, Barbara A.; Gamazon, Eric R.; Getz, Gad; Gillard, Bryan M.; Guigo, Roderic; Hambright, Kenneth W.; Hariharan, Pushpa; Hasz, Rick; Im, Hae K.; Jewell, Scott; Karasik, Ellen; Kellis, Manolis; Kheradpour, Pouya; Koester, Susan; Koller, Daphne; Konkashbaev, Anuar; Lappalainen, Tuuli; Little, Roger; Liu, Jun; Lo, Edmund; Lonsdale, John T.; Lu, Chunrong; MacArthur, Daniel G.; Magazine, Harold; Maller, Julian B.; Marcus, Yvonne; Mash, Deborah C.; McCarthy, Mark I.; McLean, Jeffrey; Mestichelli, Bernadette; Miklos, Mark; Monlong, Jean; Mosavel, Magboeba; Moser, Michael T.; Mostafavi, Sara; Nicolae, Dan L.; Pritchard, Jonathan; Qi, Liqun; Ramsey, Kimberly; Rivas, Manuel A.; Robles, Barnaby E.; Rohrer, Daniel C.; Salvatore, Mike; Sammeth, Michael; Seleski, John; Shad, Saboor; Siminoff, Laura A.; Stephens, Matthew; Struewing, Jeff; Sullivan, Timothy; Sullivan, Susan; Syron, John; Tabor, David; Taherian, Mehran; Tejada, Jorge; Temple, Gary F.; Thomas, Jeffrey A.; Thomson, Alexander W.; Tidwell, Denee; Traino, Heather M.; Tu, Zhidong; Valley, Dana R.; Volpi, Simona; Walters, Gary D.; Ward, Lucas D.; Wen, Xiaoquan; Winckler, Wendy; Wu, Shenpei; Zhu, Jun

    2015-01-01

    The Genotype-Tissue Expression (GTEx) project, sponsored by the NIH Common Fund, was established to study the correlation between human genetic variation and tissue-specific gene expression in non-diseased individuals. A significant challenge was the collection of high-quality biospecimens for

  6. Nakagami imaging for detecting thermal lesions induced by high-intensity focused ultrasound in tissue.

    Science.gov (United States)

    Rangraz, Parisa; Behnam, Hamid; Tavakkoli, Jahan

    2014-01-01

    High-intensity focused ultrasound induces focalized tissue coagulation by increasing the tissue temperature in a tight focal region. Several methods have been proposed to monitor high-intensity focused ultrasound-induced thermal lesions. Currently, ultrasound imaging techniques that are clinically used for monitoring high-intensity focused ultrasound treatment are standard pulse-echo B-mode ultrasound imaging, ultrasound temperature estimation, and elastography-based methods. On the contrary, the efficacy of two-dimensional Nakagami parametric imaging based on the distribution of the ultrasound backscattered signals to quantify properties of soft tissue has recently been evaluated. In this study, ultrasound radio frequency echo signals from ex vivo tissue samples were acquired before and after high-intensity focused ultrasound exposures and then their Nakagami parameter and scaling parameter of Nakagami distribution were estimated. These parameters were used to detect high-intensity focused ultrasound-induced thermal lesions. Also, the effects of changing the acoustic power of the high-intensity focused ultrasound transducer on the Nakagami parameters were studied. The results obtained suggest that the Nakagami distribution's scaling and Nakagami parameters can effectively be used to detect high-intensity focused ultrasound-induced thermal lesions in tissue ex vivo. These parameters can also be used to understand the degree of change in tissue caused by high-intensity focused ultrasound exposures, which could be interpreted as a measure of degree of variability in scatterer concentration in various parts of the high-intensity focused ultrasound lesion.

  7. Assessing fibrinogen extravasation into Alzheimer's disease brain using high-content screening of brain tissue microarrays.

    Science.gov (United States)

    Narayan, Pritika J; Kim, Sue-Ling; Lill, Claire; Feng, Sheryl; Faull, Richard L M; Curtis, Maurice A; Dragunow, Michael

    2015-05-30

    Tissue microarrays are commonly used to evaluate disease pathology however methods to automate and quantify pathological changes are limited. This article demonstrates the utility of the VSlide scanner (MetaSystems) for automated image acquisition from immunolabelled tissue microarray slides, and subsequent automated image analysis with MetaXpress (Molecular Devices) software to obtain objective, efficient and reproducible data from immunolabelled tissue microarray sections. Significant increases in fibrinogen immunolabelling were observed in 29 Alzheimer's disease cases compared to 28 control cases analysed from a single tissue microarray slide. Western blot analysis also demonstrated significant increases in fibrinogen immunolabelling in 6 Alzheimer's cases compared to 6 control cases. The observed changes were also validated with gold standard blinded manual H-scoring. VSlide Metafer software offers a 'tissue microarray acquisition' plugin for easy mapping of tissue cores with their original position on the tissue microarray map. High resolution VSlide images are compatible with MetaXpress image analysis software. This article details the coupling of these two technologies to accurately and reproducibly analyse immunolabelled tissue microarrays within minutes, compared to the gold standard method of manual counting using H-scores which is significantly slower and prone to inter-observer variation. Here, we couple brain tissue microarray technology with high-content screening and automated image analysis as a powerful way to address bottle necks in data generation and improve throughput, as well as sensitivity to study biological/pathological changes in brain disease. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Does Adipose Tissue Thermogenesis Play a Role in Metabolic Health?

    Directory of Open Access Journals (Sweden)

    Craig Porter

    2013-01-01

    Full Text Available The function ascribed to brown adipose tissue in humans has long been confined to thermoregulation in neonates, where this thermogenic capacity was thought lost with maturation. Recently, brown adipose tissue depots have been identified in adult humans. The significant oxidative capacity of brown adipocytes and the ability of their mitochondria to respire independently of ATP production, has led to renewed interest in the role that these adipocytes play in human energy metabolism. In our view, there is a need for robust physiological studies determining the relationship between molecular signatures of brown adipose tissue, adipose tissue mitochondrial function, and whole body energy metabolism, in order to elucidate the significance of thermogenic adipose tissue in humans. Until such information is available, the role of thermogenic adipose tissue in human metabolism and the potential that these adipocytes may prevent or treat obesity and metabolic diseases in humans will remain unknown. In this article, we summarize the recent literature pertaining to brown adipose tissue function with the aims of drawing the readers’ attention to the lack of data concerning the role of brown adipocytes in human physiology, and to the potential limitations of current research strategies.

  9. A high-resolution method for the localization of proanthocyanidins in plant tissues

    Directory of Open Access Journals (Sweden)

    Panter Stephen

    2011-05-01

    Full Text Available Abstract Background Histochemical staining of plant tissues with 4-dimethylaminocinnamaldehyde (DMACA or vanillin-HCl is widely used to characterize spatial patterns of proanthocyanidin accumulation in plant tissues. These methods are limited in their ability to allow high-resolution imaging of proanthocyanidin deposits. Results Tissue embedding techniques were used in combination with DMACA staining to analyze the accumulation of proanthocyanidins in Lotus corniculatus (L. and Trifolium repens (L. tissues. Embedding of plant tissues in LR White or paraffin matrices, with or without DMACA staining, preserved the physical integrity of the plant tissues, allowing high-resolution imaging that facilitated cell-specific localization of proanthocyanidins. A brown coloration was seen in proanthocyanidin-producing cells when plant tissues were embedded without DMACA staining and this was likely to have been due to non-enzymatic oxidation of proanthocyanidins and the formation of colored semiquinones and quinones. Conclusions This paper presents a simple, high-resolution method for analysis of proanthocyanidin accumulation in organs, tissues and cells of two plant species with different patterns of proanthocyanidin accumulation, namely Lotus corniculatus (birdsfoot trefoil and Trifolium repens (white clover. This technique was used to characterize cell type-specific patterns of proanthocyanidin accumulation in white clover flowers at different stages of development.

  10. Thermic effect of food and beta-adrenergic thermogenic responsiveness in habitually exercising and sedentary healthy adult humans.

    Science.gov (United States)

    Stob, Nicole R; Bell, Christopher; van Baak, Marleen A; Seals, Douglas R

    2007-08-01

    The thermic effect of food (TEF) is an important physiological determinant of total daily energy expenditure (EE) and energy balance. TEF is believed to be mediated in part by sympathetic nervous system activation and consequent beta-adrenergic receptor (beta-AR) stimulation of metabolism. TEF is greater in habitually exercising than in sedentary adults, despite similar postprandial sympathetic nervous system activation. We determined whether augmented TEF in habitually exercising adults is associated with enhanced peripheral thermogenic responsiveness to beta-AR stimulation. In separate experiments in 22 sedentary and 29 habitually exercising adults, we measured the increase in EE (indirect calorimetry, ventilated hood) during beta-AR stimulation (intravenous isoproterenol: 6, 12, and 24 ng x kg fat-free mass(-1) x min(-1)) and EE before and after a liquid meal (40% of resting EE; 53% carbohydrate, 32% fat, 15% protein). The increase in EE during incremental isoproterenol administration was greater (P = 0.01) in habitual exercisers (0.34 +/- 0.03, 0.54 +/- 0.04, 0.81 +/- 0.05 kJ/min; means +/- SE) than in sedentary adults (0.26 +/- 0.03, 0.40 +/- 0.03, 0.64 +/- 0.04 kJ/min). The area under the TEF response curve was also greater (P = 0.04) in habitual exercisers (160 +/- 9 kJ) than in sedentary adults (130 +/- 11 kJ) and was positively related to beta-AR thermogenic responsiveness (r = 0.32, P = 0.02). We conclude that TEF is related to beta-AR thermogenic responsiveness and that the greater TEF in habitual exercisers is attributable in part to their augmented beta-AR thermogenic responsiveness. Our results also suggest that peripheral thermogenic responsiveness to beta-AR stimulation is a physiological determinant of TEF and hence energy balance in healthy adult humans.

  11. Modeling and predicting tissue movement and deformation for high intensity focused ultrasound therapy.

    Directory of Open Access Journals (Sweden)

    Xiangyun Liao

    Full Text Available In ultrasound-guided High Intensity Focused Ultrasound (HIFU therapy, the target tissue (such as a tumor often moves and/or deforms in response to an external force. This problem creates difficulties in treating patients and can lead to the destruction of normal tissue. In order to solve this problem, we present a novel method to model and predict the movement and deformation of the target tissue during ultrasound-guided HIFU therapy.Our method computationally predicts the position of the target tissue under external force. This prediction allows appropriate adjustments in the focal region during the application of HIFU so that the treatment head is kept aligned with the diseased tissue through the course of therapy. To accomplish this goal, we utilize the cow tissue as the experimental target tissue to collect spatial sequences of ultrasound images using the HIFU equipment. A Geodesic Localized Chan-Vese (GLCV model is developed to segment the target tissue images. A 3D target tissue model is built based on the segmented results. A versatile particle framework is constructed based on Smoothed Particle Hydrodynamics (SPH to model the movement and deformation of the target tissue. Further, an iterative parameter estimation algorithm is utilized to determine the essential parameters of the versatile particle framework. Finally, the versatile particle framework with the determined parameters is used to estimate the movement and deformation of the target tissue.To validate our method, we compare the predicted contours with the ground truth contours. We found that the lowest, highest and average Dice Similarity Coefficient (DSC values between predicted and ground truth contours were, respectively, 0.9615, 0.9770 and 0.9697.Our experimental result indicates that the proposed method can effectively predict the dynamic contours of the moving and deforming tissue during ultrasound-guided HIFU therapy.

  12. Nasal associated lymphoid tissue of the Syrian golden hamster expresses high levels of PrPC.

    Directory of Open Access Journals (Sweden)

    Melissa D Clouse

    Full Text Available The key event in the pathogenesis of the transmissible spongiform encephalopathies is a template-dependent misfolding event where an infectious isoform of the prion protein (PrPSc comes into contact with native prion protein (PrPC and changes its conformation to PrPSc. In many extraneurally inoculated models of prion disease this PrPC misfolding event occurs in lymphoid tissues prior to neuroinvasion. The primary objective of this study was to compare levels of total PrPC in hamster lymphoid tissues involved in the early pathogenesis of prion disease. Lymphoid tissues were collected from golden Syrian hamsters and Western blot analysis was performed to quantify PrPC levels. PrPC immunohistochemistry (IHC of paraffin embedded tissue sections was performed to identify PrPC distribution in tissues of the lymphoreticular system. Nasal associated lymphoid tissue contained the highest amount of total PrPC followed by Peyer's patches, mesenteric and submandibular lymph nodes, and spleen. The relative levels of PrPC expression in IHC processed tissue correlated strongly with the Western blot data, with high levels of PrPC corresponding with a higher percentage of PrPC positive B cell follicles. High levels of PrPC in lymphoid tissues closely associated with the nasal cavity could contribute to the relative increased efficiency of the nasal route of entry of prions, compared to other routes of infection.

  13. Chemical sympathectomy alters food intake and thermogenic responses to catecholamines in rats.

    Science.gov (United States)

    Villanueva, Iván; Piñón, Manuel; Quevedo-Corona, Lucía; Martínez-Olivares, Rubén; Racotta, Radu

    2002-07-05

    It has been suggested that the sympathetic nervous system contributes to the short-term control of feeding. The adrenergic innervation of some splanchnic organs seems to be especially involved in such processes, since catecholamines reduce feeding only when injected intraperitoneally or intraportally. In this work, the effects of neonatal sympathetic denervation with guanethidine (Gnt) upon food intake were assessed in adult rats. Gnt-treated male rats had lower body weight gain. The hypophagic response to intraperitoneal (ip) norepinephrine was 70% higher in Gnt-treated animals as compared to controls (P food intake possibly related, in part, to the sensitization of the alpha-adrenergic porto-hepatic response to endogenous catecholamines. Compared with controls, Gnt-treated rats also showed a limited thermogenic capacity not related to feeding, and a greater degree of carbohydrate oxidation, possibly due to a defect in E-induced lipolysis, which is beta-adrenergic.

  14. Thermogenic response to adrenaline during restricted blood flow in the forearm

    DEFF Research Database (Denmark)

    Simonsen, L; Stefl, B; Christensen, N J

    1999-01-01

    To elucidate the underlying mechanism behind the thermogenic effect of adrenaline in human skeletal muscle, nine healthy subjects were studied during intravenous infusion of adrenaline. Restriction of blood flow to one forearm was obtained by external compression of the brachial artery, to separate...... a direct metabolic effect of adrenaline from an effect dependent on increased blood flow. The other arm served as the control arm. In the control arm, the forearm blood flow increased 4.7-fold (from 2.0 +/- 0.3 to 9.3 +/- 1.5 mL 100 g(-1) min(-1), P adrenaline infusion. Adrenaline...... not increase (baseline period: 5.6 +/- 2.3 micromol 100 g(-1) min(-1), adrenaline period: 6.1 +/- 2.1 micromol 100 g(-1) min(-1), P = 0.54). The experimental design and the difficulties in interpretation of the result are discussed. The results give evidence for the hypothesis that the vascular system plays...

  15. Towards high throughput tissue engineering: development of chitosan-calcium phosphate scaffolds for engineering bone tissue from embryonic stem cells.

    Science.gov (United States)

    Ko, Junghyuk; Kolehmainen, Kathleen; Ahmed, Farid; Jun, Martin Bg; Willerth, Stephanie M

    2012-01-01

    Tissue engineering strategies have shown promise for the repair of damaged organs, including bone. One of the major challenges associated with tissue engineering is how to scale up such processes for high throughput manufacturing of biomaterial scaffolds used to support stem cell culture. Generation of certain types of 3D biomaterial scaffolds, including chitosan-calcium phosphate blends, involves a slow fabrication process followed by a lengthy required freeze drying step. This work investigates the use of automated microwave vacuum drying technology as an alternative to traditional freeze drying as a method of fabricating chitosan-calcium phosphate scaffolds for supporting embryonic stem cell cultures. Scaffolds produced using both drying techniques possess similar properties when characterized using scanning electron microscopy and this paper is the first to report that both types of these scaffolds support undifferentiated embryonic stem cell culture as well as promote stem cell differentiation into osteogenic lineages when treated with the appropriate factors. Compared to existing scaffold manufacturing processes using freeze drying, the use of microwave vacuum drying will lead to faster production times while reducing the costs, enabling high-throughput manufacturing of biomaterial scaffolds for stem cell applications.

  16. Iron supplementation at high altitudes induces inflammation and oxidative injury to lung tissues in rats

    Energy Technology Data Exchange (ETDEWEB)

    Salama, Samir A., E-mail: salama.3@buckeyemail.osu.edu [High Altitude Research Center, Taif University, Al-Haweiah, Taif 21974 (Saudi Arabia); Department of Biochemistry, Faculty of Pharmacy, Al-Azhar University, Cairo 11751 (Egypt); Department of Pharmacology and GTMR Unit, College of Clinical Pharmacy, Taif University, Al-Haweiah, Taif 21974 (Saudi Arabia); Omar, Hany A. [Department of Pharmacology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62514 (Egypt); Maghrabi, Ibrahim A. [Department of Clinical Pharmacy, College of Clinical Pharmacy, Taif University, Al-Haweiah, Taif 21974 (Saudi Arabia); AlSaeed, Mohammed S. [Department of Surgery, College of Medicine, Taif University, Al-Haweiah, Taif 21974 (Saudi Arabia); EL-Tarras, Adel E. [High Altitude Research Center, Taif University, Al-Haweiah, Taif 21974 (Saudi Arabia)

    2014-01-01

    Exposure to high altitudes is associated with hypoxia and increased vulnerability to oxidative stress. Polycythemia (increased number of circulating erythrocytes) develops to compensate the high altitude associated hypoxia. Iron supplementation is, thus, recommended to meet the demand for the physiological polycythemia. Iron is a major player in redox reactions and may exacerbate the high altitudes-associated oxidative stress. The aim of this study was to explore the potential iron-induced oxidative lung tissue injury in rats at high altitudes (6000 ft above the sea level). Iron supplementation (2 mg elemental iron/kg, once daily for 15 days) induced histopathological changes to lung tissues that include severe congestion, dilatation of the blood vessels, emphysema in the air alveoli, and peribronchial inflammatory cell infiltration. The levels of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α), lipid peroxidation product and protein carbonyl content in lung tissues were significantly elevated. Moreover, the levels of reduced glutathione and total antioxidant capacity were significantly reduced. Co-administration of trolox, a water soluble vitamin E analog (25 mg/kg, once daily for the last 7 days of iron supplementation), alleviated the lung histological impairments, significantly decreased the pro-inflammatory cytokines, and restored the oxidative stress markers. Together, our findings indicate that iron supplementation at high altitudes induces lung tissue injury in rats. This injury could be mediated through excessive production of reactive oxygen species and induction of inflammatory responses. The study highlights the tissue injury induced by iron supplementation at high altitudes and suggests the co-administration of antioxidants such as trolox as protective measures. - Highlights: • Iron supplementation at high altitudes induced lung histological changes in rats. • Iron induced oxidative stress in lung tissues of rats at high altitudes. • Iron

  17. High-throughput extraction and quantification method for targeted metabolomics in murine tissues.

    Science.gov (United States)

    Zukunft, Sven; Prehn, Cornelia; Röhring, Cornelia; Möller, Gabriele; Hrabě de Angelis, Martin; Adamski, Jerzy; Tokarz, Janina

    2018-01-01

    Global metabolomics analyses using body fluids provide valuable results for the understanding and prediction of diseases. However, the mechanism of a disease is often tissue-based and it is advantageous to analyze metabolomic changes directly in the tissue. Metabolomics from tissue samples faces many challenges like tissue collection, homogenization, and metabolite extraction. We aimed to establish a metabolite extraction protocol optimized for tissue metabolite quantification by the targeted metabolomics AbsoluteIDQ™ p180 Kit (Biocrates). The extraction method should be non-selective, applicable to different kinds and amounts of tissues, monophasic, reproducible, and amenable to high throughput. We quantified metabolites in samples of eleven murine tissues after extraction with three solvents (methanol, phosphate buffer, ethanol/phosphate buffer mixture) in two tissue to solvent ratios and analyzed the extraction yield, ionization efficiency, and reproducibility. We found methanol and ethanol/phosphate buffer to be superior to phosphate buffer in regard to extraction yield, reproducibility, and ionization efficiency for all metabolites measured. Phosphate buffer, however, outperformed both organic solvents for amino acids and biogenic amines but yielded unsatisfactory results for lipids. The observed matrix effects of tissue extracts were smaller or in a similar range compared to those of human plasma. We provide for each murine tissue type an optimized high-throughput metabolite extraction protocol, which yields the best results for extraction, reproducibility, and quantification of metabolites in the p180 kit. Although the performance of the extraction protocol was monitored by the p180 kit, the protocol can be applicable to other targeted metabolomics assays.

  18. Ultra-fast processing of gigapixel Tissue MicroArray images using high performance computing.

    Science.gov (United States)

    Wang, Yinhai; McCleary, David; Wang, Ching-Wei; Kelly, Paul; James, Jackie; Fennell, Dean A; Hamilton, Peter W

    2011-10-01

    Tissue MicroArrays (TMAs) are a valuable platform for tissue based translational research and the discovery of tissue biomarkers. The digitised TMA slides or TMA Virtual Slides, are ultra-large digital images, and can contain several hundred samples. The processing of such slides is time-consuming, bottlenecking a potentially high throughput platform. A High Performance Computing (HPC) platform for the rapid analysis of TMA virtual slides is presented in this study. Using an HP high performance cluster and a centralised dynamic load balancing approach, the simultaneous analysis of multiple tissue-cores were established. This was evaluated on Non-Small Cell Lung Cancer TMAs for complex analysis of tissue pattern and immunohistochemical positivity. The automated processing of a single TMA virtual slide containing 230 patient samples can be significantly speeded up by a factor of circa 22, bringing the analysis time to one minute. Over 90 TMAs could also be analysed simultaneously, speeding up multiplex biomarker experiments enormously. The methodologies developed in this paper provide for the first time a genuine high throughput analysis platform for TMA biomarker discovery that will significantly enhance the reliability and speed for biomarker research. This will have widespread implications in translational tissue based research.

  19. Treatment of hypergranulation tissue with high potency topical corticosteroids in children.

    Science.gov (United States)

    McShane, Diana B; Bellet, Jane S

    2012-01-01

    Hypergranulation is having more granulation tissue than needed to fill a wound defect. Some pediatric dermatologists and most dermatologic surgeons will encounter this complication during their careers. Associated factors include wound site, prolonged inflammation, an imbalance in matrix metalloproteinases, and excessive angiogenesis. Reported treatments have included silver nitrate, excision, laser ablation, and topical corticosteroids. Our case series supports the use of medium- to high-potency topical corticosteroids in the treatment of hypergranulation tissue. © 2012 Wiley Periodicals, Inc.

  20. Impedance changes in liver tissue exposed in vitro to high-energy ultrasound.

    Science.gov (United States)

    Jossinet, Jacques; Trillaud, Christian; Chesnais, Sabrina

    2005-04-01

    The present study reports the impedance changes observed in bovine liver samples exposed in vitro to high-intensity ultrasound. The measurement frequency ranged from 80 kHz to 2 MHz. The treatment resulted in the average increase of 20% in impedance magnitude at low frequency and the average decrease of 30% at high frequency. The phase angle increased significantly by more than 15 degrees at all measurement frequencies. The slope of the log-modulus of impedance against log-frequency increased in treated tissue at frequencies above 500 kHz. This change was attributed to the alteration of the capacitive response of the tissue. The experimental observations are consistent with the known changes induced by high-energy ultrasound in liver tissue. This study confirmed that ultrasound energy produces measurable changes in a tissue's impedance and that indices can be derived to distinguish between original and treated tissues. The results obtained in liver tissue need confirmation in organs treatable with therapeutic ultrasound, such as breast and prostate.

  1. Quantifying Heterogeneity in Emphysema from High Resolution Computed Tomography: A Lung Tissue Research Consortium Study

    Science.gov (United States)

    Yilmaz, Cuneyt; Dane, Dan M.; Patel, Nova C.; Hsia, Connie C.W.

    2012-01-01

    Rationale and Objective To quantify spatial distribution of emphysema using high-resolution computed tomography (HRCT), we applied semi-automated analysis with internal attenuation calibration to measure regional air volume, tissue volume, and fractional tissue volume (FTV=tissue/[air+tissue] volume) in well-characterized patients studied by the Lung Tissue Research Consortium (LTRC). Methods HRCT was obtained at supine end-inspiration and end-expiration, and prone end-inspiration from 31 patients with mild, moderate, severe, or very severe emphysema (stages II–V, FEV1>75%, 51–75%, 21–50% and ≤20% predicted, respectively). Control data were from 20 healthy non-smokers (stage I). Each lobe was analyzed separately. Heterogeneity of FTV was assessed from coefficients of variation (CV) within and among lobes, and the kurtosis and skewness of FTV histograms. Results In emphysema, lobar air volume increased up to 177% except in the right middle lobe. Lobar tissue volume increased up to 107% in mild-moderate stages then normalized in advanced stages. Normally, FTV was up to 82% higher in lower than upper lobes. In mild-moderate emphysema, lobar FTV increased by up to 74% above normal at supine inspiration. In severe emphysema FTV declined below normal in all lobes and positions in correlation with pulmonary function (pemphysema distribution and heterogeneity from that in ILD. Early emphysema is associated with elevated tissue volume and FTV, consistent with hyperemia, inflammation or atelectasis. PMID:23122057

  2. Metabolic response of visceral white adipose tissue of obese mice exposed for 5 days to human room temperature compared to mouse thermoneutrality

    NARCIS (Netherlands)

    Stelt, van der Inge; Hoevenaars, Femke; Široká, Jitka; Ronde, de Lidwien; Friedeckỳ, David; Keijer, Jaap; Schothorst, van Evert

    2017-01-01

    Housing of laboratory mice at room temperature (22°C) might be considered a constant cold stress, which induces a thermogenic program in brown adipose tissue (BAT). However, the early adaptive response of white adipose tissue (WAT), the fat storage organ of the body, to a change from

  3. Proinsulin-producing, hyperglycemia-induced adipose tissue macrophages underlie insulin resistance in high fat-fed diabetic mice

    Science.gov (United States)

    Adipose tissue macrophages play an important role in the pathogenesis of obese type 2 diabetes. High-fat diet-induced obesity has been shown to lead to adipose tissue macrophages accumulation in rodents;however, the impact of hyperglycemia on adipose tissue macrophages dynamics in high-fat diet-fed ...

  4. Dissolvable base scaffolds allow tissue penetration of high-aspect-ratio flexible microneedles.

    Science.gov (United States)

    Yagi, Satoshi; Yamagiwa, Shota; Kubota, Yoshihiro; Sawahata, Hirohito; Numano, Rika; Imashioya, Tatsuya; Oi, Hideo; Ishida, Makoto; Kawano, Takeshi

    2015-09-16

    Microscale needle technology is important in electrophysiological studies, drug/chemical delivery systems, optogenetic applications, and so on. In this study, dissolvable needle-base scaffold realizes penetration of high-aspect-ratio flexible microneedles (e.g., 500 μm length) into biological tissues. This methodology, which is applicable to numerous high-aspect-ratio flexible microneedles, should reduce the invasiveness and provide safer tissue penetrations than conventional approaches. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Bottom-simulating reflector dynamics at Arctic thermogenic gas provinces: An example from Vestnesa Ridge, offshore west Svalbard

    Science.gov (United States)

    Plaza-Faverola, A.; Vadakkepuliyambatta, S.; Hong, W.-L.; Mienert, J.; Bünz, S.; Chand, S.; Greinert, J.

    2017-06-01

    The Vestnesa Ridge comprises a >100 km long sediment drift located between the western continental slope of Svalbard and the Arctic mid-ocean ridges. It hosts a deep water (>1000 m) gas hydrate and associated seafloor seepage system. Near-seafloor headspace gas compositions and its methane carbon isotopic signature along the ridge indicate a predominance of thermogenic gas sources feeding the system. Prediction of the base of the gas hydrate stability zone for theoretical pressure and temperature conditions and measured gas compositions results in an unusual underestimation of the observed bottom-simulating reflector (BSR) depth. The BSR is up to 60 m deeper than predicted for pure methane and measured gas compositions with >99% methane. Models for measured gas compositions with >4% higher-order hydrocarbons result in a better BSR approximation. However, the BSR remains >20 m deeper than predicted in a region without active seepage. A BSR deeper than predicted is primarily explained by unaccounted spatial variations in the geothermal gradient and by larger amounts of thermogenic gas at the base of the gas hydrate stability zone. Hydrates containing higher-order hydrocarbons form at greater depths and higher temperatures and contribute with larger amounts of carbons than pure methane hydrates. In thermogenic provinces, this may imply a significant upward revision (up to 50% in the case of Vestnesa Ridge) of the amount of carbon in gas hydrates.

  6. Efficacy and safety of a popular thermogenic drink after 28 days of ingestion

    Directory of Open Access Journals (Sweden)

    Stout Jeffrey R

    2008-10-01

    Full Text Available Abstract Background We have recently demonstrated that consuming a thermogenic drink (TD acutely increases energy expenditure and serum markers of lipolysis in healthy, college-aged individuals. The purpose of this study was to determine if consuming TD over 28 days affects its acute thermogenic and lipolytic effects as well as body composition and clinical chemistry safety markers. Methods Sixty healthy, males (mean ± SE; 23 ± 1 years, 177 ± 2 cm, 81.7 ± 2.1 kg, 22.8 ± 1.4% body fat; n = 30 and females (23 ± 1 years, 166 ± 2 cm, 62.1 ± 1.8 kg, 28.3 ± 1.4% body fat; n = 30 reported to the laboratory on day 0 (T1 for determination of body composition, resting energy expenditure (REE as well as glycerol and free fatty acid (FFA levels before and after ingesting either 336 ml of TD or a non-caloric, non-caffeinated placebo (PLA drink. Following day 0, participants supplemented daily with 336 ml·day-1 of either TD or PLA and repeated identical testing procedures on day 28 (T2. Day 28 area under the curve (AUC values were calculated for REE, FFA, and glycerol. Day 28 acute data and prolonged AUC comparisons between groups were analyzed using ANOVAs with repeated measures. Results Percent body fat (p = 0.02 and fat mass (p = 0.01 decreased in the TD group compared to the PLA group after 28 days. Day 28 FFA AUC values (p = 0.048 were greater in the TD group compared to the PLA group. There was no significant difference in day 28 REE AUC values (p = 0.30 or glycerol AUC values (p = 0.21, although a significant increase in REE values in the PLA group may have confounded these findings. There were no differences between groups concerning blood and clinical safety markers. Conclusion Within-group elevations in FFA and REE values in the TD group were still evident following a 28-day supplementation period which may contribute to the observed decrements in %BF. Further, prolonged TD supplementation did not alter the assessed clinical safety markers

  7. Incidence of Staphylococcus aureus nasal colonization and soft tissue infection among high school football players.

    Science.gov (United States)

    Lear, Aaron; McCord, Gary; Peiffer, Jeffrey; Watkins, Richard R; Parikh, Arpan; Warrington, Steven

    2011-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) skin and soft tissue infections have been documented with increasing frequency in both team and individual sports in recent years. It also seems that the level of MRSA skin and soft tissue infections in the general population has increased. One hundred ninety athletes from 6 local high school football teams were recruited for this prospective observational study to document nasal colonization and the potential role this plays in skin and soft tissue infections in football players and, in particular, MRSA infections. Athletes had nasal swabs done before their season started, and they filled out questionnaires regarding potential risk factors for skin and soft tissue infections. Those enrolled in the study were then observed over the course of the season for skin and soft tissue infections. Those infected had data about their infections collected. One hundred ninety of 386 available student athletes enrolled in the study. Forty-four of the subjects had nasal colonization with methicillin-susceptible S. aureus, and none were colonized with MRSA. There were 10 skin and soft tissue infections (8 bacterial and 2 fungal) documented over the course of the season. All were treated as outpatients with oral or topical antibiotics, and none were considered serious. Survey data from the preseason questionnaire showed 21% with skin infection, 11% with methicillin-susceptible S. aureus, and none with MRSA infection during the past year. Three reported a remote history of MRSA infection. We documented an overall skin infection rate of 5.3% among high school football players over a single season. Our results suggest that skin and soft tissue infection may not be widespread among high school athletes in northeast Ohio.

  8. High-Resolution Ultrasound-Switchable Fluorescence Imaging in Centimeter-Deep Tissue Phantoms with High Signal-To-Noise Ratio and High Sensitivity via Novel Contrast Agents.

    Science.gov (United States)

    Cheng, Bingbing; Bandi, Venugopal; Wei, Ming-Yuan; Pei, Yanbo; D'Souza, Francis; Nguyen, Kytai T; Hong, Yi; Yuan, Baohong

    2016-01-01

    For many years, investigators have sought after high-resolution fluorescence imaging in centimeter-deep tissue because many interesting in vivo phenomena-such as the presence of immune system cells, tumor angiogenesis, and metastasis-may be located deep in tissue. Previously, we developed a new imaging technique to achieve high spatial resolution in sub-centimeter deep tissue phantoms named continuous-wave ultrasound-switchable fluorescence (CW-USF). The principle is to use a focused ultrasound wave to externally and locally switch on and off the fluorophore emission from a small volume (close to ultrasound focal volume). By making improvements in three aspects of this technique: excellent near-infrared USF contrast agents, a sensitive frequency-domain USF imaging system, and an effective signal processing algorithm, for the first time this study has achieved high spatial resolution (~ 900 μm) in 3-centimeter-deep tissue phantoms with high signal-to-noise ratio (SNR) and high sensitivity (3.4 picomoles of fluorophore in a volume of 68 nanoliters can be detected). We have achieved these results in both tissue-mimic phantoms and porcine muscle tissues. We have also demonstrated multi-color USF to image and distinguish two fluorophores with different wavelengths, which might be very useful for simultaneously imaging of multiple targets and observing their interactions in the future. This work has opened the door for future studies of high-resolution centimeter-deep tissue fluorescence imaging.

  9. Dynamic changes in high and low mammographic density human breast tissues maintained in murine tissue engineering chambers during various murine peripartum states and over time.

    Science.gov (United States)

    Chew, G L; Huang, D; Huo, C W; Blick, T; Hill, P; Cawson, J; Frazer, H; Southey, M D; Hopper, J L; Henderson, M A; Haviv, I; Thompson, E W

    2013-07-01

    Mammographic density (MD) is a strong heritable risk factor for breast cancer, and may decrease with increasing parity. However, the biomolecular basis for MD-associated breast cancer remains unclear, and systemic hormonal effects on MD-associated risk is poorly understood. This study assessed the effect of murine peripartum states on high and low MD tissue maintained in a xenograft model of human MD. Method High and low MD human breast tissues were precisely sampled under radiographic guidance from prophylactic mastectomy specimens of women. The high and low MD tissues were maintained in separate vascularised biochambers in nulliparous or pregnant SCID mice for 4 weeks, or mice undergoing postpartum involution or lactation for three additional weeks. High and low MD biochamber material was harvested for histologic and radiographic comparisons during various murine peripartum states. High and low MD biochamber tissues in nulliparous mice were harvested at different timepoints for histologic and radiographic comparisons. Results High MD biochamber tissues had decreased stromal (p = 0.0027), increased adipose (p = 0.0003) and a trend to increased glandular tissue areas (p = 0.076) after murine postpartum involution. Stromal areas decreased (p = 0.042), while glandular (p = 0.001) and adipose areas (p = 0.009) increased in high MD biochamber tissues during lactation. A difference in radiographic density was observed in high (p = 0.0021) or low MD biochamber tissues (p = 0.004) between nulliparous, pregnant and involution groups. No differences in tissue composition were observed in high or low MD biochamber tissues maintained for different durations, although radiographic density increased over time. Conclusion High MD biochamber tissues had measurable histologic changes after postpartum involution or lactation. Alterations in radiographic density occurred in biochamber tissues between different peripartum states and over time. These findings

  10. Laser-generated ultrasound for high-precision cutting of tissue-mimicking gels (Conference Presentation)

    Science.gov (United States)

    Lee, Taehwa; Luo, Wei; Li, Qiaochu; Guo, L. Jay

    2017-03-01

    Laser-generated focused ultrasound has shown great promise in precisely treating cells and tissues by producing controlled micro-cavitation within the acoustic focal volume (30 MPa, negative pressure amplitude). By moving cavitation spots along pre-defined paths through a motorized stage, tissue-mimicking gels of different elastic moduli were cut into different shapes (rectangle, triangle, and circle), leaving behind the same shape of holes, whose sizes are less than 1 mm. The cut line width is estimated to be less than 50 um (corresponding to localized cavitation region), allowing for accurate cutting. This novel approach could open new possibility for in-vivo treatment of diseased tissues in a high-precision manner (i.e., high-precision invisible sonic scalpel).

  11. Effects of Tamoxifen and oestrogen on histology and radiographic density in high and low mammographic density human breast tissues maintained in murine tissue engineering chambers.

    Science.gov (United States)

    Chew, G L; Huo, C W; Huang, D; Blick, T; Hill, P; Cawson, J; Frazer, H; Southey, M C; Hopper, J L; Britt, K; Henderson, M A; Haviv, I; Thompson, E W

    2014-11-01

    Mammographic density (MD) is a strong risk factor for breast cancer. It is altered by exogenous endocrine treatments, including hormone replacement therapy and Tamoxifen. Such agents also modify breast cancer (BC) risk. However, the biomolecular basis of how systemic endocrine therapy modifies MD and MD-associated BC risk is poorly understood. This study aims to determine whether our xenograft biochamber model can be used to study the effectiveness of therapies aimed at modulating MD, by examine the effects of Tamoxifen and oestrogen on histologic and radiographic changes in high and low MD tissues maintained within the biochamber model. High and low MD human tissues were precisely sampled under radiographic guidance from prophylactic mastectomy fresh specimens of high-risk women, then inserted into separate vascularized murine biochambers. The murine hosts were concurrently implanted with Tamoxifen, oestrogen or placebo pellets, and the high and low MD biochamber tissues maintained in the murine host environment for 3 months, before the high and low MD biochamber tissues were harvested for histologic and radiographic analyses. The radiographic density of high MD tissue maintained in murine biochambers was decreased in Tamoxifen-treated mice compared to oestrogen-treated mice (p = 0.02). Tamoxifen treatment of high MD tissue in SCID mice led to a decrease in stromal (p = 0.009), and an increase in adipose (p = 0.023) percent areas, compared to placebo-treated mice. No histologic or radiographic differences were observed in low MD biochamber tissue with any treatment. High MD biochamber tissues maintained in mice implanted with Tamoxifen, oestrogen or placebo pellets had dynamic and measurable histologic compositional and radiographic changes. This further validates the dynamic nature of the MD xenograft model, and suggests the biochamber model may be useful for assessing the underlying molecular pathways of Tamoxifen-reduced MD, and in testing of other

  12. Transcriptomic and proteomic profiling of two porcine tissues using high-throughput technologies

    Directory of Open Access Journals (Sweden)

    Panitz Frank

    2009-01-01

    Full Text Available Abstract Background The recent development within high-throughput technologies for expression profiling has allowed for parallel analysis of transcriptomes and proteomes in biological systems such as comparative analysis of transcript and protein levels of tissue regulated genes. Until now, such studies of have only included microarray or short length sequence tags for transcript profiling. Furthermore, most comparisons of transcript and protein levels have been based on absolute expression values from within the same tissue and not relative expression values based on tissue ratios. Results Presented here is a novel study of two porcine tissues based on integrative analysis of data from expression profiling of identical samples using cDNA microarray, 454-sequencing and iTRAQ-based proteomics. Sequence homology identified 2.541 unique transcripts that are detectable by both microarray hybridizations and 454-sequencing of 1.2 million cDNA tags. Both transcript-based technologies showed high reproducibility between sample replicates of the same tissue, but the correlation across these two technologies was modest. Thousands of genes being differentially expressed were identified with microarray. Out of the 306 differentially expressed genes, identified by 454-sequencing, 198 (65% were also found by microarray. The relationship between the regulation of transcript and protein levels was analyzed by integrating iTRAQ-based proteomics data. Protein expression ratios were determined for 354 genes, of which 148 could be mapped to both microarray and 454-sequencing data. A comparison of the expression ratios from the three technologies revealed that differences in transcript and protein levels across heart and muscle tissues are positively correlated. Conclusion We show that the reproducibility within cDNA microarray and 454-sequencing is high, but that the agreement across these two technologies is modest. We demonstrate that the regulation of transcript

  13. The influence of high temperature on the possibility of DNA typing in various human tissues.

    Science.gov (United States)

    Maciejewska, Agnieszka; Wlodarczyk, Renata; Pawlowski, Ryszard

    2015-01-01

    The identification of unknown victims of high temperatures (fire, terrorist attack, and other disasters) is one of the most difficult tasks faced by forensic geneticists. The main aim of this study was to in­vestigate the availability of DNA isolated from various human tissue samples exposed to high temperatures of 100–1000°C for 5 and 10 minutes. Samples of varying thickness of thigh muscle, liver, heart, adipose tissue, bone, teeth, hair and nails of 52 fresh cadavers and 59 healthy teeth of 29 volunteers were used. The study was performed using the following commercially available STR (Short Tandem Repeats) and miniSTR kits: AmpFlSTR®SGM Plus® and AmpFlSTR®MiniFilerTM. Hyper variable region I (HVI) of human mitochondrial DNA (mtDNA) was sequenced with BigDye Terminator Cycle Sequencing Kit 1.1. The PEP (Primer-Extension Preamplification) method was used for the whole human genome amplification. It was possible to obtain complete DNA profiles (AmpFlSTR®SGM Plus®, AmpFlSTR®MiniFilerTM Applied Biosystems, USA and mtDNA HVI region) for tissue samples of heart, liver and thigh muscle, exposed up to 900°C for 5 min. However, under the applied conditions, limited usefulness of hair, nails and teeth for identification purposes was shown. DNA stability in tissues subjected to incineration depends on many factors, like tissue type and its thickness, temperature and time of exposure. In the cases of human remains exposed to high temperatures, samples of soft tissues of the highest weight (thickness) provide the best chance of successful identification through the genetic analysis. In some cases of negative results, even if using mtDNA typing, application of the whole genome amplification (WGA) technique could provide the expected results for highly degraded DNA templates.

  14. Creating High-Resolution Multiscale Maps of Human Tissue Using Multi-beam SEM.

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    André F Pereira

    2016-11-01

    Full Text Available Multi-beam scanning electron microscopy (mSEM enables high-throughput, nano-resolution imaging of macroscopic tissue samples, providing an unprecedented means for structure-function characterization of biological tissues and their cellular inhabitants, seamlessly across multiple length scales. Here we describe computational methods to reconstruct and navigate a multitude of high-resolution mSEM images of the human hip. We calculated cross-correlation shift vectors between overlapping images and used a mass-spring-damper model for optimal global registration. We utilized the Google Maps API to create an interactive map and provide open access to our reconstructed mSEM datasets to both the public and scientific communities via our website www.mechbio.org. The nano- to macro-scale map reveals the tissue's biological and material constituents. Living inhabitants of the hip bone (e.g. osteocytes are visible in their local extracellular matrix milieu (comprising collagen and mineral and embedded in bone's structural tissue architecture, i.e. the osteonal structures in which layers of mineralized tissue are organized in lamellae around a central blood vessel. Multi-beam SEM and our presented methodology enable an unprecedented, comprehensive understanding of health and disease from the molecular to organ length scale.

  15. Chinese highly pathogenic porcine reproductive and respiratory syndrome virus exhibits more extensive tissue tropism for pigs

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    Li Limin

    2012-09-01

    Full Text Available Abstract Background The highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV emerging in China exhibits high fatality to pigs. However, the mechanism related to the increased pathogenicity of the virus remains unclear. In the present study, the differences in tissue tropism between the highly pathogenic PRRSV strain (JXwn06 and the low pathogenic PRRSV strain (HB-1/3.9 were investigated using PRRSV-specific immunohistochemistry (IHC staining to provide evidence for elucidating possible mechanism of the pathogenicity of Chinese highly pathogenic PRRSV. Findings IHC examination showed that PRRSV antigen in the tissues including spleen, tonsil, thymus, kidney, cerebellum, stomach, small intestine, large intestine, turbinal bone and laryngeal cartilage was positive in more pigs inoculated with JXwn06 than HB-1/3.9, and the tissues including trachea, esophagus, liver, mandibular gland and thyroid gland were positive for viral antigen in the pigs inoculated with JXwn06, but not in the pigs inoculated with HB-1/3.9. Meanwhile, we observed that epithelium in tissues including interlobular bile duct in liver, distal renal tubule of kidney, esophageal gland and tracheal gland were positive for viral antigen only in JXwn06-inoculated pigs, and epithelium of gastric mucosa and fundic gland, and intestinal gland were positive for viral antigen in both JXwn06- and HB-1/3.9-inoculated pigs, using monoclonal antibodies to N and Nsp2 proteins. Conclusions Taken together, these findings indicate that the highly pathogenic PRRSV JXwn06 displays an expanded tissue tropism in vivo, suggesting this may contribute to its high pathogenicity to pigs.

  16. Mechanical high-intensity focused ultrasound destruction of soft tissue: working mechanisms and physiologic effects

    NARCIS (Netherlands)

    Hoogenboom, M.; Eikelenboom, D.C.; Brok, M.H.M.G.M. den; Heerschap, A.; Futterer, J.J.; Adema, G.J.

    2015-01-01

    The best known method of high-intensity focused ultrasound is thermal ablation, but interest in non-thermal, mechanical destruction is increasing. The advantages of mechanical ablation are that thermal protein denaturation remains limited and less damage is created to the surrounding tissue by

  17. Quantifying Biochemical Alterations in Brown and Subcutaneous White Adipose Tissues of Mice Using Fourier Transform Infrared Widefield Imaging

    Directory of Open Access Journals (Sweden)

    Ebrahim Aboualizadeh

    2017-05-01

    Full Text Available Stimulating increased thermogenic activity in adipose tissue is an important biological target for obesity treatment, and label-free imaging techniques with the potential to quantify stimulation-associated biochemical changes to the adipose tissue are highly sought after. In this study, we used spatially resolved Fourier transform infrared (FTIR imaging to quantify biochemical changes caused by cold exposure in the brown and subcutaneous white adipose tissues (BAT and s-WAT of 6 week-old C57BL6 mice exposed to 30°C (N = 5, 24°C (N = 5, and 10°C (N = 5 conditions for 10 days. Fat exposed to colder temperatures demonstrated greater thermogenic activity as indicated by increased messenger RNA expression levels of a panel of thermogenic marker genes including uncoupling protein 1 (UCP-1 and Dio2. Protein to lipid ratio, calculated from the ratio of the integrated area from 1,600 to 1,700 cm−1 (amide I to the integrated area from 2,830 to 2,980 cm−1 (saturated lipids, was elevated in 10°C BAT and s-WAT compared to 24°C (p = 0.004 and p < 0.0001 and 30°C (p = 0.0033 and p < 0.0001. Greater protein to lipid ratio was associated with greater UCP-1 expression level in the BAT (p = 0.021 and s-WAT (p = 0.032 and greater Dio2 expression in s-WAT (p = 0.033. The degree of unsaturation, calculated from the ratio of the integrated area from 2,992 to 3,020 cm−1 (unsaturated lipids to the integrated area from 2,830 to 2,980 cm−1 (saturated lipids, showed stepwise decreases going from colder-exposed to warmer-exposed BAT. Complementary 1H NMR measurements confirmed the findings from this ratio in BAT. Principal component analysis applied to FTIR spectra revealed pronounced differences in overall spectral characteristics between 30, 24, and 10°C BAT and s-WAT. Spatially resolved FTIR imaging is a promising technique to quantify cold-induced biochemical changes in BAT and s-WAT in a label-free manner.

  18. An Automated Platform for High-Resolution Tissue Imaging Using Nanospray Desorption Electrospray Ionization Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Lanekoff, Ingela T.; Heath, Brandi S.; Liyu, Andrey V.; Thomas, Mathew; Carson, James P.; Laskin, Julia

    2012-10-02

    An automated platform has been developed for acquisition and visualization of mass spectrometry imaging (MSI) data using nanospray desorption electrospray ionization (nano-DESI). The new system enables robust operation of the nano-DESI imaging source over many hours. This is achieved by controlling the distance between the sample and the probe by mounting the sample holder onto an automated XYZ stage and defining the tilt of the sample plane. This approach is useful for imaging of relatively flat samples such as thin tissue sections. Custom software called MSI QuickView was developed for visualization of large data sets generated in imaging experiments. MSI QuickView enables fast visualization of the imaging data during data acquisition and detailed processing after the entire image is acquired. The performance of the system is demonstrated by imaging rat brain tissue sections. High resolution mass analysis combined with MS/MS experiments enabled identification of lipids and metabolites in the tissue section. In addition, high dynamic range and sensitivity of the technique allowed us to generate ion images of low-abundance isobaric lipids. High-spatial resolution image acquired over a small region of the tissue section revealed the spatial distribution of an abundant brain metabolite, creatine, in the white and gray matter that is consistent with the literature data obtained using magnetic resonance spectroscopy.

  19. Ex Vivo Characterization of Canine Liver Tissue Viscoelasticity Following High Intensity Focused Ultrasound (HIFU) Ablation

    Science.gov (United States)

    Shahmirzadi, Danial; Hou, Gary Y.; Chen, Jiangang; Konofagou, Elisa E.

    2014-01-01

    Elasticity imaging has shown great promise in detecting High Intensity Focused Ultrasound (HIFU) lesions based on their distinct biomechanical properties. However, quantitative mechanical properties of the tissue and the optimal intensity for obtaining the best contrast parameters remain scarce. In this study, fresh canine livers were ablated using combinations of ISPTA intensities of 5.55, 7.16 and 9.07 kW/cm2 and time durations of 10 and 30 s ex vivo; leading to six groups of ablated tissues. Biopsy samples were then interrogated using dynamic shear mechanical testing within the range of 0.1-10 Hz to characterize the post-ablation tissue viscoelastic properties. All mechanical parameters were found to be frequency dependent. Compared to the unablated cases, all six groups of ablated tissues showed statistically-significant higher complex shear modulus and shear viscosity. However, among the ablated groups, both complex shear modulus and shear viscosity were found to monotonically increase in groups 1-4 (5.55 kW/cm2 for 10 s, 7.16 kW/cm2 for 10 s, 9.07 kW/cm2 & 10 s, and 5.55 kW/cm2 & 30 s, respectively), but decrease in groups 5 and 6 (7.16 kW/cm2 for 30 s, and 9.07 kW/cm2 for 30 s, respectively). For groups 5 and 6, the temperature was expected to exceed the boiling point, and therefore, the decreased stiffening could be due to the compromised integrity of the tissue microstructure. Future studies are needed to estimate the tissue mechanical properties in vivo and perform real-time monitoring of tissue alterations during ablation. PMID:24315395

  20. Quantifying heterogeneity in emphysema from high-resolution computed tomography: a lung tissue research consortium study.

    Science.gov (United States)

    Yilmaz, Cuneyt; Dane, Dan M; Patel, Nova C; Hsia, Connie C W

    2013-02-01

    To quantify spatial distribution of emphysema using high-resolution computed tomography (HRCT), we applied semiautomated analysis with internal attenuation calibration to measure regional air volume, tissue volume, and fractional tissue volume (FTV = tissue/[air + tissue] volume) in well-characterized patients studied by the Lung Tissue Research Consortium (LTRC). HRCT was obtained at supine end-inspiration and end-expiration, and prone end-inspiration from 31 patients with mild, moderate, severe, or very severe emphysema (stages II-V, forced expiratory volume at 1 second >75%, 51%-75%, 21%-50% and ≤20% predicted, respectively). Control data were from 20 healthy non-smokers (stage I). Each lobe was analyzed separately. Heterogeneity of FTV was assessed from coefficients of variation (CV) within and among lobes, and the kurtosis and skewness of FTV histograms. In emphysema, lobar air volume increased up to 177% above normal except in the right middle lobe. Lobar tissue volume increased up to 107% in mild-moderate stages then normalized in advanced stages. Normally, FTV was up to 82% higher in lower than upper lobes. In mild-moderate emphysema, lobar FTV increased by up to 74% above normal at supine inspiration. In severe emphysema, FTV declined below normal in all lobes and positions in correlation with pulmonary function (P < .05). Markers of FTV heterogeneity increased steadily with disease stage in correlation with pulmonary function (P < .05); the pattern is distinct from that seen in interstitial lung disease (ILD). CT-derived biomarkers differentiate the spatial patterns of emphysema distribution and heterogeneity from that in ILD. Early emphysema is associated with elevated tissue volume and FTV, consistent with hyperemia, inflammation or atelectasis. Published by Elsevier Inc.

  1. High-intensity-focused-ultrasound (HIFU) induced homeostasis and tissue ablation

    Science.gov (United States)

    Chauhan, Sunita; Michel, M. S.; Alken, Peter; Kohrmann, K. U.; Haecker, Axel

    2003-06-01

    At high intensity levels, ultrasound energy focused into remote tissue targets in human body has shown to produce thermal necrosis in circumscribed regions with sub-millimeter accuracy. The non-invasive modality known as HIFU has enormous potential for thermal ablation of cancers/tumors of the human body without any adverse effects in the surrounding normal tissue. In this paper, empirical results for parametric assessment and interdependence of several exposure variables are presented for producing thermal necrosis as well as hemostasis. Multiple HIFU transducers in selective spatial configuration have been deployed using a suitably designed experiemntal harness, with and without motorized jig scanning. The pre-planning and on-line procedure for treatment and specified instrumentation is described. Custom designed 25mm aperture HIFU probes resonating at 2 MHz focused at 64 and 80 mm are used. Results have been obtained in ex-vivo animal tissue and in vitro biological phantoms for hemostasis.

  2. High resolution ultrasonic scanning of animal and human tissue in-vivo

    Energy Technology Data Exchange (ETDEWEB)

    Roswell, R. L.; Goans, R. E.; Cantrell, Jr., J. H.

    1977-08-01

    Burns impose one of the most serious injuries to the skin due to the organ's function within the body system and to the body as a whole. In an effort to better deal with the burn wound by the immediate excision and grafting of third degree burns, a high resolution (approximately 0.2 mm) ultrasonic pulse-echo technique was developed for determining burn depth. The experimental subjects were Yorkshire pigs because of the histological similarity between human and porcine skin. Burn depths were readily identifiable immediately postburn with the ultrasonic techniques, as were general trends concerning the burn-viable and viable-fat interfaces. The tissue characteristics, density and acoustic attenuation, effecting the impedance mismatch at the burn-viable tissue interface were investigated. The methods of fluid displacements and specific gravities yielded density values, while spectrum analyses produced attenuation measurements for normal, viable and burned tissue samples.

  3. Vasoconstrictor effect of high FFA/albumin ratios in adipose tissue in vivo

    DEFF Research Database (Denmark)

    Bülow, J; Madsen, J; Astrup, A

    1985-01-01

    as well as in young dogs after this treatment. The administration of Intralipid did not per se induce the vasoconstriction. The vasoconstriction took place simultaneously with increasing FFA/albumin molar ratios. The results support our previous findings in perfused fat pads that high molar FFA......Subcutaneous or perirenal adipose tissue blood flow was measured with the 133Xe-washout technique before and after intravenous injection or infusion of Intralipid in six anesthetized, otherwise intact mongrel dogs. In four anesthetized mongrel puppies adipose tissue blood flow was measured....../albumin ratios increase vascular resistance in adipose tissue and they give further support to our suggestion that this vasoconstriction may be a link in a negative-feedback mechanism regulating FFA-mobilization in relation to FFA utilization....

  4. High intensity interval training improves liver and adipose tissue insulin sensitivity

    Science.gov (United States)

    Marcinko, Katarina; Sikkema, Sarah R.; Samaan, M. Constantine; Kemp, Bruce E.; Fullerton, Morgan D.; Steinberg, Gregory R.

    2015-01-01

    Objective Endurance exercise training reduces insulin resistance, adipose tissue inflammation and non-alcoholic fatty liver disease (NAFLD), an effect often associated with modest weight loss. Recent studies have indicated that high-intensity interval training (HIIT) lowers blood glucose in individuals with type 2 diabetes independently of weight loss; however, the organs affected and mechanisms mediating the glucose lowering effects are not known. Intense exercise increases phosphorylation and inhibition of acetyl-CoA carboxylase (ACC) by AMP-activated protein kinase (AMPK) in muscle, adipose tissue and liver. AMPK and ACC are key enzymes regulating fatty acid metabolism, liver fat content, adipose tissue inflammation and insulin sensitivity but the importance of this pathway in regulating insulin sensitivity with HIIT is unknown. Methods In the current study, the effects of 6 weeks of HIIT were examined using obese mice with serine–alanine knock-in mutations on the AMPK phosphorylation sites of ACC1 and ACC2 (AccDKI) or wild-type (WT) controls. Results HIIT lowered blood glucose and increased exercise capacity, food intake, basal activity levels, carbohydrate oxidation and liver and adipose tissue insulin sensitivity in HFD-fed WT and AccDKI mice. These changes occurred independently of weight loss or reductions in adiposity, inflammation and liver lipid content. Conclusions These data indicate that HIIT lowers blood glucose levels by improving adipose and liver insulin sensitivity independently of changes in adiposity, adipose tissue inflammation, liver lipid content or AMPK phosphorylation of ACC. PMID:26909307

  5. [Phospholipids and structural modification of tissues and cell membranes for adaptation in high altitude mountains].

    Science.gov (United States)

    Iakovlev, V M; Vishnevskiĭ, A A; Shanazarov, A S

    2012-01-01

    The nature of the impact of physical factors of high altitudes (3200 m) on the lipids of tissues and membranes of animals was researched. It was established that the adaptation process in Wistar rats was followed by peroxide degradation and subsequent modification of the phospholipids' structure of tissues and microsomal membranes. Adaptive phospholipids reconstruction takes place in microsomal membranes in the tissues of the lungs, brain, liver and skeletal muscles. Together with this, the amount of phosphatidylinositol and phosphatidic acid accumulates, indicating that the hydrolysis of phosphatidylinositol-4, 5 biphosphate to diacylglycerol and secondary messenger--inositol triphosphate, occurs. A decrease in temperature adaptation (+10 degrees C) leads to a more noticeable shift in peroxide oxidation of lipids, phospholipid structure in the tissues and membranes rather than adaptation in thermoneutral conditions (+30 degrees C). Modification of lipid composition of tissues and cell membranes in the highlands obviously increases the adaptive capabilities of cells of the whole body: physical performance and resistance to hypoxia increases in animals.

  6. Growth hormone-binding proteins in high-speed cytosols of multiple tissues of the rabbit.

    Science.gov (United States)

    Herington, A C; Ymer, S; Roupas, P; Stevenson, J

    1986-04-11

    Soluble, specific binding protein(s) for growth hormone (GH) have been identified and partially characterized in high-speed cytosolic preparations from a number of rabbit tissues. The binding of 125I-labelled human GH to proteins in liver, heart, adipose tissue, skeletal muscle and kidney cytosols was dependent on time and cytosolic protein concentration. By Scatchard analysis, the binding affinities (KA: (2-7) X 10(9) M-1) were somewhat higher than those generally reported for membrane GH receptors. The binding proteins had a greater specificity for somatotrophic hormones than lactogenic hormones, although the kidney appeared to have, in addition, a lactogen-binding protein. By gel filtration, the Mr of the cytosolic GH-binding protein was approximately 100 000 in all tissues. None of the binding proteins was detectable by the poly(ethylene glycol) precipitation method used widely for soluble hormone receptors. The cytosolic GH-binding proteins also cross-reacted with a monoclonal antibody to the rabbit liver membrane GH receptor. These results indicate the ubiquitous presence of apparently naturally soluble GH-binding proteins in the cytosolic fractions of several tissues in the rabbit. Of great interest is their presence in muscle, where GH receptors or binding proteins have not previously been detected, despite muscle being recognized as a classical GH target tissue.

  7. Kaiso is highly expressed in TNBC tissues of women of African ancestry compared to Caucasian women.

    Science.gov (United States)

    Bassey-Archibong, Blessing I; Hercules, Shawn M; Rayner, Lyndsay G A; Skeete, Desiree H A; Smith Connell, Suzanne P; Brain, Ian; Daramola, Adetola; Banjo, Adekunbiola A F; Byun, Jung S; Gardner, Kevin; Dushoff, Jonathan; Daniel, Juliet M

    2017-11-01

    Triple-negative breast cancer (TNBC) is most prevalent in young women of African ancestry (WAA) compared to women of other ethnicities. Recent studies found a correlation between high expression of the transcription factor Kaiso, TNBC aggressiveness, and ethnicity. However, little is known about Kaiso expression and localization patterns in TNBC tissues of WAA. Herein, we analyze Kaiso expression patterns in TNBC tissues of African (Nigerian), Caribbean (Barbados), African American (AA), and Caucasian American (CA) women. Formalin-fixed and paraffin embedded (FFPE) TNBC tissue blocks from Nigeria and Barbados were utilized to construct a Nigerian/Barbadian tissue microarray (NB-TMA). This NB-TMA and a commercially available TMA comprising AA and CA TNBC tissues (AA-CA-YTMA) were subjected to immunohistochemistry to assess Kaiso expression and subcellular localization patterns, and correlate Kaiso expression with TNBC clinical features. Nigerian and Barbadian women in our study were diagnosed with TNBC at a younger age than AA and CA women. Nuclear and cytoplasmic Kaiso expression was observed in all tissues analyzed. Analysis of Kaiso expression in the NB-TMA and AA-CA-YTMA revealed that nuclear Kaiso H scores were significantly higher in Nigerian, Barbadian, and AA women compared with CA women. However, there was no statistically significant difference in nuclear Kaiso expression between Nigerian versus Barbadian women, or Barbadian versus AA women. High levels of nuclear Kaiso expression were detected in patients with a higher degree of African heritage compared to their Caucasian counterparts, suggesting a role for Kaiso in TNBC racial disparity.

  8. The cold-induced lipokine 12,13-diHOME promotes fatty acid transport into brown adipose tissue

    DEFF Research Database (Denmark)

    Lynes, Matthew D; Leiria, Luiz O; Lundh, Morten

    2017-01-01

    and glucose tolerance; as a class, these lipids are referred to as 'lipokines'. Because BAT is a specialized metabolic tissue that takes up and burns lipids and is linked to systemic metabolic homeostasis, we hypothesized that there might be thermogenic lipokines that activate BAT in response to cold. Here we...

  9. Acute effects of a thermogenic nutritional supplement on cycling time to exhaustion and muscular strength in college-aged men

    Directory of Open Access Journals (Sweden)

    Beck Travis W

    2009-07-01

    Full Text Available Abstract Background The purpose of the present study was to examine the acute effects of a thermogenic nutritional supplement containing caffeine, capsaicin, bioperine, and niacin on muscular strength and endurance performance. Methods Twenty recreationally-active men (mean ± SD age = 21.5 ± 1.4 years; stature = 178.2 ± 6.3 cm; mass = 76.5 ± 9.9 kg; VO2 PEAK = 3.05 ± 0.59 L/min-1 volunteered to participate in this randomized, double-blinded, placebo-controlled, cross-over study. All testing took place over a three-week period, with each of the 3 laboratory visits separated by 7 days (± 2 hours. During the initial visit, a graded exercise test was performed on a Lode Corival cycle ergometer (Lode, Groningen, Netherlands until exhaustion (increase of 25 W every 2 min to determine the maximum power output (W at the VO2 PEAK (Parvo Medics TrueOne® 2400 Metabolic Measurement System, Sandy, Utah. In addition, one-repetition maximum (1-RM strength was assessed using the bench press (BP and leg press (LP exercises. During visits 2 and 3, the subjects were asked to consume a capsule containing either the active supplement (200 mg caffeine, 33.34 mg capsaicin, 5 mg bioperine, and 20 mg niacin or the placebo (175 mg of calcium carbonate, 160 mg of microcrystalline cellulose, 5 mg of stearic acid, and 5 mg of magnesium stearate in an identical capsule 30 min prior to the testing. Testing included a time-to-exhaustion (TTE ride on a cycle ergometer at 80% of the previously-determined power output at VO2 PEAK followed by 1-RM LP and BP tests. Results There were no differences (p > 0.05 between the active and placebo trials for BP, LP, or TTE. However, for the BP and LP scores, the baseline values (visit 1 were less than the values recorded during visits 2 and 3 (p ≤ 0.05. Conclusion Our findings indicated that the active supplement containing caffeine, capsaicin, bioperine, and niacin did not alter muscular strength or cycling endurance when compared

  10. Differential modulation of cytosolic lipases activities in liver and adipose tissue by high-carbohydrate diets.

    Science.gov (United States)

    Rodrigues, Angélica Heringer; Moreira, Carolina Campos Lima; Mario, Érica Guilhen; de Souza Cordeiro, Letícia Maria; Avelar, Gleide Fernandes; Botion, Leida Maria; Chaves, Valéria Ernestânia

    2016-08-01

    Several studies have demonstrated that a high-fructose (FRUC) diet induces metabolic and haemodynamic abnormalities, known as the metabolic syndrome, which are characterised by obesity, glucose intolerance, insulin resistance, dyslipidaemia and hypertension. In this study, the effect of a FRUC diet (60 % fructose) for 8 weeks on the metabolism of lipids in liver and epididymal adipose tissue from Wistar rats was compared with the AIN-93M diet and the effects of the AIN-93M diet were compared with a chow diet. The FRUC diet induced marked increases in both hepatocyte lipid droplet volume and postprandial serum levels of triacylglycerol (TAG), but reduced the postprandial serum levels of insulin. The AIN-93M diet induced marked increases in the hepatocyte lipid droplet volume and the serum levels of insulin, without affecting the serum levels of TAG. We found that isocaloric substitution of cornstarch, dextrinised cornstarch and sucrose (AIN-93M diet) for fructose did not affect the hepatic VLDL-TAG secretion and adipose tissue glucose uptake, lipolysis and cytosolic lipases activities in rats. However, the high-fructose diet induced a severe steatosis in liver accompanied by a decrease in cytosolic lipases activities. In adipose tissue, the FRUC diet induced a decrease in the lipoprotein lipase activity, and an increase in lipogenesis. FRUC and AIN-93M diets induced changes in lipid homeostasis in liver and adipose tissue by distinct biochemical mechanisms.

  11. Continuous-specimen-flow, high-throughput, 1-hour tissue processing. A system for rapid diagnostic tissue preparation.

    Science.gov (United States)

    Morales, Azorides R; Essenfeld, Harold; Essenfeld, Ervin; Duboue, Maria Carmen; Vincek, Vladimir; Nadji, Mehrdad

    2002-05-01

    Current conventional tissue-processing methods employ fixation of tissues with neutral buffered formalin, dehydration with alcohol, and clearing with xylene before paraffin impregnation. Because the time required for this procedure is usually 8 hours or longer, it is customary to process tissues in automated instruments throughout the night. Although this time-honored method continues to serve histology laboratories well, it has a number of shortcomings, such as a 1-day delay of diagnosis, the need to batch specimens, the relatively large volumes and toxicity of reagents used, and the extent of RNA degradation. To describe a rapid new method of tissue processing using a continuous-throughput technique. Design.-We used a combination of common histologic reagents, excluding formalin and xylene, as well as microwave energy, to develop a rapid processing method. The effect of this method on the quality of histomorphology, histochemistry, immunohistochemistry, and RNA content of processed tissue was compared with that of adjacent tissue sections processed by the conventional processing technique. We also assessed the impact of this rapid processing system on our practice by comparing the turnaround times of surgical pathology reports before and after its implementation. The new processing method permitted preparation of paraffin blocks from fresh or prefixed tissue in about 1 hour. The procedure allowed continuous flow of specimens at 15-minute intervals. It eliminated the use of formalin and xylene in the processing and used considerably lower volumes of other chemical reagents. Histomorphologic, histochemical, and immunohistochemical results were comparable to the parallel sections prepared by the conventional method. The new technique, however, preserved higher quality RNA. Use of the new methodology led to the diagnosis and reporting of more than one third of surgical pathology specimens on the same day that they were received, as compared to 1% of same

  12. Tandem high-pressure freezing and quick freeze substitution of plant tissues for transmission electron microscopy.

    Science.gov (United States)

    Bobik, Krzysztof; Dunlap, John R; Burch-Smith, Tessa M

    2014-10-13

    Since the 1940s transmission electron microscopy (TEM) has been providing biologists with ultra-high resolution images of biological materials. Yet, because of laborious and time-consuming protocols that also demand experience in preparation of artifact-free samples, TEM is not considered a user-friendly technique. Traditional sample preparation for TEM used chemical fixatives to preserve cellular structures. High-pressure freezing is the cryofixation of biological samples under high pressures to produce very fast cooling rates, thereby restricting ice formation, which is detrimental to the integrity of cellular ultrastructure. High-pressure freezing and freeze substitution are currently the methods of choice for producing the highest quality morphology in resin sections for TEM. These methods minimize the artifacts normally associated with conventional processing for TEM of thin sections. After cryofixation the frozen water in the sample is replaced with liquid organic solvent at low temperatures, a process called freeze substitution. Freeze substitution is typically carried out over several days in dedicated, costly equipment. A recent innovation allows the process to be completed in three hours, instead of the usual two days. This is typically followed by several more days of sample preparation that includes infiltration and embedding in epoxy resins before sectioning. Here we present a protocol combining high-pressure freezing and quick freeze substitution that enables plant sample fixation to be accomplished within hours. The protocol can readily be adapted for working with other tissues or organisms. Plant tissues are of special concern because of the presence of aerated spaces and water-filled vacuoles that impede ice-free freezing of water. In addition, the process of chemical fixation is especially long in plants due to cell walls impeding the penetration of the chemicals to deep within the tissues. Plant tissues are therefore particularly challenging, but

  13. Activation of the immune system incurs energetic costs but has no effect on the thermogenic performance of house sparrows during acute cold challenge.

    Science.gov (United States)

    King, Marisa O; Swanson, David L

    2013-06-01

    Trade-offs between the immune system and other condition-dependent life-history traits (reproduction, predator avoidance and somatic growth) have been well documented in both birds and mammals. However, no studies have examined the impact of immune activation on thermoregulatory performance during acute cold exposure. Because of their high surface-area-to-volume ratios, small birds incur high energetic costs associated with thermoregulation during cold exposure. Consequently, we predicted that the immune system and the thermoregulatory system would compete for energetic resources. To test this, we immunologically challenged adult house sparrows (Passer domesticus) with 5 mg kg(-1) of lipopolysaccharide (LPS) to induce an acute phase response and measured both resting (RMR; minimum metabolic rate) and summit ( ; maximal metabolic rate during cold exposure) metabolic rates. We found that birds injected with LPS had significantly higher RMR and than birds injected with phosphate-buffered saline, indicating that LPS-treated birds were able to support the cost of both immune activation and thermoregulation under conditions eliciting maximal thermogenic performance. These results suggest that, in the absence of a pathogen, birds that experience short-term activation of the immune system have higher energetic costs during cold exposure, but immune activation does not compromise maximum thermoregulatory performance.

  14. Monoamine reuptake site occupancy of sibutramine: Relationship to antidepressant-like and thermogenic effects in rats.

    Science.gov (United States)

    Li, Yu-Wen; Langdon, Shaun; Pieschl, Rick; Strong, Todd; Wright, Robert N; Rohrbach, Kenneth; Lelas, Snjezana; Lodge, Nicholas J

    2014-08-15

    Sibutramine was formerly marketed as an anti-obesity agent. The current study investigated the relationships between monoamine reuptake site occupancy for sibutramine and both its antidepressant-like efficacy and thermogenic effects. Sibutramine's effects on locomotor activity (LMA) and food intake were also evaluated. Sibutramine occupied monoamine reuptake binding sites with the rank order of potency of NET>SERT>DAT; at 10mg/kg, po, occupancy was 95% NET, 81% SERT and 73% DAT. Sibutramine produced antidepressant-like behavior in the forced swim test; at the lowest effective dose (3mg/kg, po) occupancy was 61%, 90% and 23% at SERT, NET and DAT sites, respectively. Sibutramine also increased body core temperature in a dose- and time-dependent manner; at the lowest effective dose (30mg/kg) SERT, NET and DAT occupancies were respectively 78%, 86% and 59%. A significant decrease in food consumption was observed at 3 and 10mg/kg, po. LMA was increased at ≥10mg/kg, sc. The relationship between efficacy in the FST and occupancy was also determined for citalopram, fluoxetine and reboxetine. Similarly, the relationship between thermogenesis and target occupancy for several single or double/triple reuptake inhibitors was measured and showed that >40-50% DAT binding was required for thermogenesis. Thermogenesis was blocked by the D1 antagonist SCH39166 (3mg/kg, sc). Our findings indicate that the antidepressant-like effect of sibutramine may result from additive or synergistic actions on the three reuptake binding targets. At higher doses, sibutramine produces thermogenesis; DAT inhibition and activation of dopamine D1 receptors are required for this effect. Published by Elsevier B.V.

  15. Hypothalamic and hindbrain melanocortin receptors contribute to the feeding, thermogenic, and cardiovascular action of melanocortins.

    Science.gov (United States)

    Skibicka, Karolina P; Grill, Harvey J

    2009-12-01

    Forebrain ventricular delivery of melanocortin receptor (MC3/4R) agonist increases energy expenditure and decreases food intake (FI). Because forebrain ventricular delivery provides ligand to various anatomically distributed MC3/4R-bearing nuclei, it is unclear which of the receptor subpopulations contributes to the feeding suppression and the sympathetic-thermogenic effects observed. The literature indicates that reexpression of MC4R in the paraventricular nucleus (PVH) affects the feeding but not the energetic phenotype of the MC4R knockout, suggesting that divergent MC4R populations mediate energy expenditure (hindbrain) and FI (hypothalamus) effects of stimulation. Not consistent with this view are data indicating that PVH sympathetic projection neurons express MC4Rs and that feeding effects are induced from hindbrain MC4R sites. Therefore, we hypothesize an opposing perspective: that stimulation of anatomically diverse MC3/4R-bearing nuclei triggers energetic as well as feeding effects. To test this hypothesis, ventricle subthreshold doses of MC3/4R agonist (5 and 10 pmol) were applied in separate experiments to six hindbrain and hypothalamic sites; core temperature (Tc), heart rate (HR), spontaneous activity (SPA), and FI were measured in behaving rats. Nucleus tractus solitarius and PVH stimulation increased Tc, HR, and SPA and decreased FI. Rostral ventrolateral medulla, parabrachial nucleus, and retrochiasmatic area stimulation increased Tc, HR, but not SPA, and decreased FI. The response profile differed to some extent for each nucleus tested, suggesting differential output circuitries for the measured parameters. Data are consistent with the view that energetic and feeding responses are not controlled by regionally divergent MC3/4Rs and can be elicited from multiple, anatomically distributed MC3/4R populations.

  16. Increased thermogenic response to food and fat oxidation in female athletes: relationship with VO(2 max).

    Science.gov (United States)

    López, P; Ledoux, M; Garrel, D R

    2000-09-01

    The thermogenic response to food (TRF) and substrate oxidation were studied in 12 endurance-trained and 13 untrained female subjects. Energy expenditure and substrate oxidation were calculated by indirect calorimetry before and for 6 h after an oral test meal and after the same meal given intragastrically on a separate occasion. The TRF was calculated after the oral meal, the obligatory component after the intragastric meal (OTRF), and the facultative component from the difference between the two. VO(2 max) was measured on a treadmill and body composition by underwater weighing. The TRF and OTRF were significantly higher in trained than in untrained subjects: 223 +/- 63 vs. 185 +/- 50 kJ/6 h (P < 0.03) and 174 +/- 38 vs. 131 +/- 37 kJ/6 h (P < 0.01) for the TRF and OTRF in trained vs. untrained subjects, respectively. Multiple regression analysis showed that maximum O(2) consumption (VO(2 max)), but not percentage of body fat, was significantly related to OTRF (r =0.68, P < 0.01). Trained subjects had higher fatty acid oxidation than untrained subjects before (0.6 vs. 0.4 mg. kg(-1). min(-1), P < 0.05) and after the oral meal (13 +/- 6 vs. 8 +/- 4 g/6 h P < 0.05). These results demonstrate that 1) TRF is higher in trained than in untrained women; 2) this is due to a higher cost of nutrient digestion, absorption and storage; 3) the difference is related to higher VO(2 max); and 4) fatty acid oxidation is greater in trained women in both the postabsorptive and postprandial states. These observations suggest that endurance training induces metabolic changes that favor leanness.

  17. Resting metabolic rate and thermogenic effect of food in vegetarian diets compared with Mediterranean diets.

    Science.gov (United States)

    Bissoli, L; Armellini, F; Zamboni, M; Mandragona, R; Ballarin, A; Bosello, O

    1999-01-01

    The purpose of our work is to compare the resting metabolic rate (RMR) and thermogenic effect of food (TEF) in a group of vegetarians and a group of subjects consuming a Mediterranean diet. The composition of the diets was similar. Thirty-two subjects were studied: 16 vegetarians (age 34 +/- 9 years, BMI 21 +/- 2) and 16 omnivors (age 30 +/- 5 years, BMI 22 +/- 3). All were in excellent general health. Each subject consumed a dish of pasta (100 g) and bread (30 g) after RMR had been measured. TEF was measured over the next 3 h and calculated as the incremental area above RMR. Energy (vegetarians and omnivors 7,727 +/- 3,516 vs 8,970 +/- 2,273 kJ/day, respectively) and carbohydrate (vegetarians and omnivors 285.1 +/- 141.3 vs. 300.1 +/- 74 g/day, respectively) intakes of the 2 groups were similar. The vegetarian group consumed a higher quantity of fiber (30.5 +/- 16.7 vs. 16.5 +/- 7.9) and a lower amount of protein (44.9 +/- 18.3 vs. 70.1 +/- 14.9) than the omnivorous group. No significant differences were observed in RMR (4.23 +/- 0.96 vs. 4.06 +/- 0.54 kJ/min) and TEF (0.50 +/- 0.25 vs. 0.38 +/- 0.25 Delta kJ/min) between the groups. Results did not change after correcting for weight, age and gender. Our study failed to show any significant differences in RMR and TEF between vegetarians and subjects consuming a Mediterranean diet. We conclude that vegetarianism per se is not accompanied by a difference in RMR and TEF when the carbohydrate content of the diet is similar to a control group of nonvegetarians.

  18. Efficient, high-speed ablation of soft tissue with few-microjoule, femtosecond pulse bursts

    CERN Document Server

    Kerse, Can; Kalaycıoğlu, Hamit; Aşık, Mehmet D; Akçaalan, Önder; Ilday, F Ömer

    2014-01-01

    Femtosecond pulses hold great promise for high-precision tissue removal. However, ablation rates are severely limited by the need to keep average laser power low to avoid collateral damage due to heat accumulation. Furthermore, previously reported pulse energies preclude delivery in flexible fibers, hindering in vivo operation. Both of these problems can be addressed through use of groups of high-repetition-rate pulses, or bursts. Here, we report a novel fiber laser and demonstrate ultrafast burst-mode ablation of brain tissue at rates approaching 1 mm$^3$/min, an order of magnitude improvement over previous reports. Burst mode operation is shown to be superior in terms of energy required and avoidance of thermal effects, compared to uniform repetition rates. These results can pave the way to in vivo operation at medically relevant speeds, delivered via flexible fibers to surgically hard-to-reach targets, or with simultaneous magnetic resonance imaging.

  19. Niacin increases adiponectin and decreases adipose tissue inflammation in high fat diet-fed mice.

    Directory of Open Access Journals (Sweden)

    Desiree Wanders

    Full Text Available To determine the effects of niacin on adiponectin and markers of adipose tissue inflammation in a mouse model of obesity.Male C57BL/6 mice were placed on a control or high-fat diet (HFD and were maintained on such diets for the duration of the study. After 6 weeks on the control or high fat diets, vehicle or niacin treatments were initiated and maintained for 5 weeks. Identical studies were conducted concurrently in HCA2 (-/- (niacin receptor(-/- mice.Niacin increased serum concentrations of the anti-inflammatory adipokine, adiponectin by 21% in HFD-fed wild-type mice, but had no effect on lean wild-type or lean or HFD-fed HCA2 (-/- mice. Niacin increased adiponectin gene and protein expression in the HFD-fed wild-type mice only. The increases in adiponectin serum concentrations, gene and protein expression occurred independently of changes in expression of PPARγ C/EBPα or SREBP-1c (key transcription factors known to positively regulate adiponectin gene transcription in the adipose tissue. Further, niacin had no effect on adipose tissue expression of ERp44, Ero1-Lα, or DsbA-L (key ER chaperones involved in adiponectin production and secretion. However, niacin treatment attenuated HFD-induced increases in adipose tissue gene expression of MCP-1 and IL-1β in the wild-type HFD-fed mice. Niacin also reduced the expression of the pro-inflammatory M1 macrophage marker CD11c in HFD-fed wild-type mice.Niacin treatment attenuates obesity-induced adipose tissue inflammation through increased adiponectin and anti-inflammatory cytokine expression and reduced pro-inflammatory cytokine expression in a niacin receptor-dependent manner.

  20. High-dose-rate (HDR) brachytherapy for the treatment of benign obstructive endobronchial granulation tissue.

    Science.gov (United States)

    Madu, Chika N; Machuzak, Michael S; Sterman, Daniel H; Musani, Ali; Ahya, Vivek; McDonough, James; Metz, James M

    2006-12-01

    Severe airway obstruction can occur in the setting of benign granulation tissue forming at bronchial anastomotic sites after lung transplantation in up to 20% of patients. Many of these benign lesions respond to stent placement, laser ablation, or balloon bronchoplasty. However, in certain cases, proliferation of granulation tissue may persist despite all therapeutic attempts. This study describes a series of refractory patients treated with high-dose-rate (HDR) brachytherapy for benign proliferation of granulation tissue, causing airway compromise. Between April 2002 and June 2005, 5 patients with significant airway compromise from recurrent granulation tissue were treated with HDR brachytherapy. All patients had previously failed to maintain a patent airway despite multiple bronchoscopic interventions. Treatment was delivered using an HDR brachytherapy afterloader with (192)Ir. Dose prescription was to a depth of 1 cm. All patients were treated weekly, with total doses ranging from 10 Gy to 21 Gy in two to three fractions. The median follow-up was 12 months. All patients experienced a reduction in therapeutic bronchoscopic procedures after HDR brachytherapy compared with the pretreatment period. With the exception of possible radiation-induced bronchitis in 1 patient, there were no other treatment related complications. At the time of this report, 2 patients have died and the other 3 are alive with marked symptomatic improvement and reduced bronchoscopic procedures. High-dose-rate brachytherapy is an effective treatment for benign proliferation of granulation tissue causing airway obstruction. The early response to therapy is encouraging and further follow-up is necessary to determine long-term durability and late effects.

  1. Niacin Increases Adiponectin and Decreases Adipose Tissue Inflammation in High Fat Diet-Fed Mice

    Science.gov (United States)

    Wanders, Desiree; Graff, Emily C.; White, B. Douglas; Judd, Robert L.

    2013-01-01

    Aims To determine the effects of niacin on adiponectin and markers of adipose tissue inflammation in a mouse model of obesity. Materials and Methods Male C57BL/6 mice were placed on a control or high-fat diet (HFD) and were maintained on such diets for the duration of the study. After 6 weeks on the control or high fat diets, vehicle or niacin treatments were initiated and maintained for 5 weeks. Identical studies were conducted concurrently in HCA2−/− (niacin receptor−/−) mice. Results Niacin increased serum concentrations of the anti-inflammatory adipokine, adiponectin by 21% in HFD-fed wild-type mice, but had no effect on lean wild-type or lean or HFD-fed HCA2−/− mice. Niacin increased adiponectin gene and protein expression in the HFD-fed wild-type mice only. The increases in adiponectin serum concentrations, gene and protein expression occurred independently of changes in expression of PPARγ C/EBPα or SREBP-1c (key transcription factors known to positively regulate adiponectin gene transcription) in the adipose tissue. Further, niacin had no effect on adipose tissue expression of ERp44, Ero1-Lα, or DsbA-L (key ER chaperones involved in adiponectin production and secretion). However, niacin treatment attenuated HFD-induced increases in adipose tissue gene expression of MCP-1 and IL-1β in the wild-type HFD-fed mice. Niacin also reduced the expression of the pro-inflammatory M1 macrophage marker CD11c in HFD-fed wild-type mice. Conclusions Niacin treatment attenuates obesity-induced adipose tissue inflammation through increased adiponectin and anti-inflammatory cytokine expression and reduced pro-inflammatory cytokine expression in a niacin receptor-dependent manner. PMID:23967184

  2. Neuromuscular manifestations of viscoelastic tissue degradation following high and low risk repetitive lumbar flexion.

    Science.gov (United States)

    Solomonow, M

    2012-04-01

    Cumulative lumbar disorder is common in individuals engaged in long term performance of repetitive and static occupational/sports activities with the spine. The triggering source and of the disorder, the tissues involved in the failure and the biomechanical, neuromuscular, and biological processes active in the initiation and development of the disorder are not known. The hypothesis is forwarded that static and repetitive (cyclic) lumbar flexion-extension and the associated repeated stretch of the various viscoelastic tissues (ligaments, fascia, facet capsule, discs, etc.) causes micro-damage in their collagen fibers followed by an acute inflammation, triggering pain and reflexive muscle spasms/hyper-excitability. Continued exposure to activities, over time, converts the acute inflammation into a chronic one, viscoelastic tissues remodeling/degeneration, modified motor control strategy and permanent disability. Changes in lumbar stability are expected during the development of the disorder. A series of experimental data from in-vivo feline is reviewed and integrated with supporting evidence from the literature to gain a valuable insight into the multi-factorial development of the disorder. Prolonged cyclic lumbar flexion-extension at high loads, high velocities, many repetitions and short in between rest periods induced transient creep/laxity in the spine, muscle spasms and reduced stability followed, several hours later, by an acute inflammation/tissue degradation, muscular hyper-excitability and increased stability. The major findings assert that viscoelastic tissues sub-failure damage is the source and inflammation is the process which governs the mechanical and neuromuscular characteristic symptoms of the disorder. A comprehensive model of the disorder is presented. The experimental data validates the hypothesis as well as provide insights into the development of potential treatment and prevention of the disorder. Copyright © 2011 Elsevier Ltd. All rights

  3. Highly porous scaffolds of PEDOT:PSS for bone tissue engineering.

    Science.gov (United States)

    Guex, Anne Géraldine; Puetzer, Jennifer L; Armgarth, Astrid; Littmann, Elena; Stavrinidou, Eleni; Giannelis, Emmanuel P; Malliaras, George G; Stevens, Molly M

    2017-10-15

    Conjugated polymers have been increasingly considered for the design of conductive materials in the field of regenerative medicine. However, optimal scaffold properties addressing the complexity of the desired tissue still need to be developed. The focus of this study lies in the development and evaluation of a conductive scaffold for bone tissue engineering. In this study PEDOT:PSS scaffolds were designed and evaluated in vitro using MC3T3-E1 osteogenic precursor cells, and the cells were assessed for distinct differentiation stages and the expression of an osteogenic phenotype. Ice-templated PEDOT:PSS scaffolds presented high pore interconnectivity with a median pore diameter of 53.6±5.9µm and a total pore surface area of 7.72±1.7m(2)·g(-1). The electrical conductivity, based on I-V curves, was measured to be 140µS·cm(-1) with a reduced, but stable conductivity of 6.1µS·cm(-1) after 28days in cell culture media. MC3T3-E1 gene expression levels of ALPL, COL1A1 and RUNX2 were significantly enhanced after 4weeks, in line with increased extracellular matrix mineralisation, and osteocalcin deposition. These results demonstrate that a porous material, based purely on PEDOT:PSS, is suitable as a scaffold for bone tissue engineering and thus represents a promising candidate for regenerative medicine. Tissue engineering approaches have been increasingly considered for the repair of non-union fractions, craniofacial reconstruction or large bone defect replacements. The design of complex biomaterials and successful engineering of 3-dimensional tissue constructs is of paramount importance to meet this clinical need. Conductive scaffolds, based on conjugated polymers, present interesting candidates to address the piezoelectric properties of bone tissue and to induce enhanced osteogenesis upon implantation. However, conductive scaffolds have not been investigated in vitro in great measure. To this end, we have developed a highly porous, electrically conductive scaffold

  4. Protection of Lotus Seedpod Proanthocyanidins on Organs and Tissues under High-intensity Excercise.

    Science.gov (United States)

    Mengyan, Zhang

    2015-01-01

    Lotus seedpod proanthocyanidins (LSPC) as a kind of polyphenols is widely used in medicines, cosmetics, health products. High-intensity exercise can cause damage to the body's organs and tissues. Different doses of LSPC is given to mice to check the function of protect effect to the body's organs and tissues under high-intensity exercise. The hemoglobin (HB) content, red blood cell (RBC) number and white blood cell (WBC) number were tested for mice after exercise. The activity of superoxide dismutase (SOD) and the contents of glutathione (GSH) and malondialdehyde (MDA) in muscle and viscera were evaluated. The result showed that LSPC can effectively reduce inflammation reaction in the body of mice with high intensity exercise, alleviate oxidative stress-induced injury of tissues and organs, and execute protective function on skeletal muscle and cardiac muscle. And the LSPC could enhance myocardial anti-oxygen and enzymatic activity which suggests the protective effects of resveratrol against exercise-induced myocardial damage in mice.

  5. Determination of vinblastine in tumour tissue with liquid chromatography-high resolution mass spectrometry.

    Science.gov (United States)

    Kosjek, Tina; Dolinšek, Tanja; Gramec, Darja; Heath, Ester; Strojan, Primož; Serša, Gregor; Čemažar, Maja

    2013-11-15

    There are virtually no analytical methods that describe determination of vinblastine and other vinca alkaloids in tumour tissue, albeit quantitative data on tumour drug amount is essential for maximal benefit of a particular anticancer treatment. The analytical method presented herein uses state-of-the-art sample preparation, separation and detection techniques to allow sensitive and selective determination of vinblastine in tumour tissue. After cryogenic grinding and sonication, tumour suspensions were extracted by Oasis MAX solid phase extraction and analysed for vinblastine with ultra-high performance liquid chromatography coupled to positive electrospray ionisation-high resolution mass spectrometric detection. The developed analytical method quantifies vinblastine down to 23 ng/g tumour tissue and shows satisfactory linearity (r(2)>0.99), precision (1.1-8.2%), accuracy (98%) and high selectivity with almost complete absence of matrix effects. The proposed method was found suitable to follow vinblastine levels in mice tumours and could be used to support preclinical pharmacologic studies. © 2013 Elsevier B.V. All rights reserved.

  6. Voluntary exercise improves murine dermal connective tissue status in high-fat diet-induced obesity.

    Science.gov (United States)

    Lőrincz, Kende; Haluszka, Dóra; Kiss, Norbert; Gyöngyösi, Nóra; Bánvölgyi, András; Szipőcs, Róbert; Wikonkál, Norbert M

    2017-04-01

    Obesity is a risk factor for several cardiovascular and metabolic diseases. Its influence on the skin is less obvious, yet certain negative effects of adipose tissue inflammation on the dermis have been suggested. Excess weight is closely associated with sedentary behavior, so any increase in physical activity is considered beneficial against obesity. To investigate the effects of obesity and physical exercise on the skin, we established a mouse model in which mice were kept either on a high-fat diet or received standard chow. After the two groups achieved a significant weight difference, physical exercise was introduced to both. Animals were given the opportunity to perform voluntary exercise for 40 min daily in a hamster wheel for a period of 8 weeks. We evaluated the status of the dermis at the beginning and at the end of the exercise period by in vivo nonlinear microscopy. Obese mice kept on high-fat diet lost weight steadily after they started to exercise. In the high-fat diet group, we could detect significantly larger adipocytes and a thicker layer of subcutaneous tissue; both changes started to normalize after exercise. Nonlinear microscopy revealed an impaired collagen structure in obese mice that improved considerably after physical activity was introduced. With the ability to detect damage on collagen structure, we set out to address the question whether this process is reversible. With the use of a novel imaging method, we were able to show the reversibility of connective tissue deterioration as a benefit of physical exercise.

  7. High-resolution NMR spectroscopy of human body fluids and tissues in relation to prostate cancer.

    Science.gov (United States)

    Kumar, Virendra; Dwivedi, Durgesh K; Jagannathan, Naranamangalam R

    2014-01-01

    High-resolution NMR spectroscopic studies of prostate tissue extracts, prostatic fluid, seminal fluid, serum and urine can be used for the detection of prostate cancer, based on the differences in their metabolic profiles. Useful diagnostic information is obtained by the detection or quantification of as many metabolites as possible and comparison with normal samples. Only a few studies have shown the potential of high-resolution in vitro NMR of prostate tissues. A survey of the literature has revealed that studies on body fluids, such as urine and serum, in relation to prostate cancer are rare. In addition, the potential of NMR of nuclei other than (1)H, such as (13)C and (31)P, has not been exploited fully. The metabolomic analysis of metabolites, detected by high-resolution NMR, may help to identify metabolites which could serve as useful biomarkers for prostate cancer detection. Such NMR-derived biomarkers would not only help in prostate cancer detection and in understanding the in vivo MRS metabolic profile, but also to investigate the biochemical and metabolic changes associated with cancer. Here, we review the published research work on body fluids in relation to prostate and prostate tissue extracts, and highlight the potential of such studies for future work. Copyright © 2013 John Wiley & Sons, Ltd.

  8. Forager bees (Apis mellifera) highly express immune and detoxification genes in tissues associated with nectar processing.

    Science.gov (United States)

    Vannette, Rachel L; Mohamed, Abbas; Johnson, Brian R

    2015-11-09

    Pollinators, including honey bees, routinely encounter potentially harmful microorganisms and phytochemicals during foraging. However, the mechanisms by which honey bees manage these potential threats are poorly understood. In this study, we examine the expression of antimicrobial, immune and detoxification genes in Apis mellifera and compare between forager and nurse bees using tissue-specific RNA-seq and qPCR. Our analysis revealed extensive tissue-specific expression of antimicrobial, immune signaling, and detoxification genes. Variation in gene expression between worker stages was pronounced in the mandibular and hypopharyngeal gland (HPG), where foragers were enriched in transcripts that encode antimicrobial peptides (AMPs) and immune response. Additionally, forager HPGs and mandibular glands were enriched in transcripts encoding detoxification enzymes, including some associated with xenobiotic metabolism. Using qPCR on an independent dataset, we verified differential expression of three AMP and three P450 genes between foragers and nurses. High expression of AMP genes in nectar-processing tissues suggests that these peptides may contribute to antimicrobial properties of honey or to honey bee defense against environmentally-acquired microorganisms. Together, these results suggest that worker role and tissue-specific expression of AMPs, and immune and detoxification enzymes may contribute to defense against microorganisms and xenobiotic compounds acquired while foraging.

  9. High-resolution ex vivo magnetic resonance angiography: a feasibility study on biological and medical tissues

    Directory of Open Access Journals (Sweden)

    Boel Lene WT

    2010-03-01

    Full Text Available Abstract Background In biomedical sciences, ex vivo angiography is a practical mean to elucidate vascular structures three-dimensionally with simultaneous estimation of intravascular volume. The objectives of this study were to develop a magnetic resonance (MR method for ex vivo angiography and to compare the findings with computed tomography (CT. To demonstrate the usefulness of this method, examples are provided from four different tissues and species: the human placenta, a rice field eel, a porcine heart and a turtle. Results The optimal solution for ex vivo MR angiography (MRA was a compound containing gelatine (0.05 g/mL, the CT contrast agent barium sulphate (0.43 mol/L and the MR contrast agent gadoteric acid (2.5 mmol/L. It was possible to perform angiography on all specimens. We found that ex vivo MRA could only be performed on fresh tissue because formalin fixation makes the blood vessels permeable to the MR contrast agent. Conclusions Ex vivo MRA provides high-resolution images of fresh tissue and delineates fine structures that we were unable to visualise by CT. We found that MRA provided detailed information similar to or better than conventional CTA in its ability to visualize vessel configuration while avoiding interfering signals from adjacent bones. Interestingly, we found that vascular tissue becomes leaky when formalin-fixed, leading to increased permeability and extravascular leakage of MR contrast agent.

  10. High resolution ultrasound elastomicroscopy imaging of soft tissues: system development and feasibility

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Y P [Rehabilitation Engineering Center, Hong Kong Polytechnic University, Kowloon, Hong Kong (China); Bridal, S L [Laboratoire d' Imagerie Parametrique, UMR CNRS 7623, University of Paris VI, Paris (France); Shi, J [Rehabilitation Engineering Center, Hong Kong Polytechnic University, Kowloon, Hong Kong (China); Saied, A [Laboratoire d' Imagerie Parametrique, UMR CNRS 7623, University of Paris VI, Paris (France); Lu, M H [Rehabilitation Engineering Center, Hong Kong Polytechnic University, Kowloon, Hong Kong (China); Jaffre, B [Laboratoire d' Imagerie Parametrique, UMR CNRS 7623, University of Paris VI, Paris (France); Mak, A F T [Rehabilitation Engineering Center, Hong Kong Polytechnic University, Kowloon, Hong Kong (China); Laugier, P [Laboratoire d' Imagerie Parametrique, UMR CNRS 7623, University of Paris VI, Paris (France)

    2004-09-07

    Research in elasticity imaging typically relies on 1-10 MHz ultrasound. Elasticity imaging at these frequencies can provide strain maps with a resolution in the order of millimetres, but this is not sufficient for applications to skin, articular cartilage or other fine structures. We developed a prototype high resolution elastomicroscopy system consisting of a 50 MHz ultrasound backscatter microscope system and a calibrated compression device using a load cell to measure the pressure applied to the specimen, which was installed between a rigidly fixed face-plate and a specimen platform. Radiofrequency data were acquired in a B-scan format (10 mm wide x 3 mm deep) in specimens of mouse skin and bovine patellar cartilage. The scanning resolution along the B-scan plane direction was 50 {mu}m, and the ultrasound signals were digitized at 500 MHz to achieve a sensitivity better than 1 {mu}m for the axial displacement measurement. Because of elevated attenuation of ultrasound at high frequencies, special consideration was necessary to design a face-plate permitting efficient ultrasound transmission into the specimen and relative uniformity of the compression. Best results were obtained using a thin plastic film to cover a specially shaped slit in the face-plate. Local tissue strain maps were constructed by applying a cross-correlation tracking method to signals obtained at the same site at different compression levels. The speed of sound in the tissue specimen (1589.8 {+-} 7.8 m s{sup -1} for cartilage and 1532.4 {+-} 4.4 m s{sup -1} for skin) was simultaneously measured during the compression test. Preliminary results demonstrated that this ultrasound elastomicroscopy technique was able to map deformations of the skin and articular cartilage specimens to high resolution, in the order of 50 {mu}m. This system can also be potentially used for the assessment of other biological tissues, bioengineered tissues or biomaterials with fine structures.

  11. High resolution ultrasound elastomicroscopy imaging of soft tissues: system development and feasibility

    Science.gov (United States)

    Zheng, Y. P.; Bridal, S. L.; Shi, J.; Saied, A.; Lu, M. H.; Jaffre, B.; Mak, A. F. T.; Laugier, P.

    2004-09-01

    Research in elasticity imaging typically relies on 1-10 MHz ultrasound. Elasticity imaging at these frequencies can provide strain maps with a resolution in the order of millimetres, but this is not sufficient for applications to skin, articular cartilage or other fine structures. We developed a prototype high resolution elastomicroscopy system consisting of a 50 MHz ultrasound backscatter microscope system and a calibrated compression device using a load cell to measure the pressure applied to the specimen, which was installed between a rigidly fixed face-plate and a specimen platform. Radiofrequency data were acquired in a B-scan format (10 mm wide × 3 mm deep) in specimens of mouse skin and bovine patellar cartilage. The scanning resolution along the B-scan plane direction was 50 µm, and the ultrasound signals were digitized at 500 MHz to achieve a sensitivity better than 1 µm for the axial displacement measurement. Because of elevated attenuation of ultrasound at high frequencies, special consideration was necessary to design a face-plate permitting efficient ultrasound transmission into the specimen and relative uniformity of the compression. Best results were obtained using a thin plastic film to cover a specially shaped slit in the face-plate. Local tissue strain maps were constructed by applying a cross-correlation tracking method to signals obtained at the same site at different compression levels. The speed of sound in the tissue specimen (1589.8 ± 7.8 m s-1 for cartilage and 1532.4 ± 4.4 m s-1 for skin) was simultaneously measured during the compression test. Preliminary results demonstrated that this ultrasound elastomicroscopy technique was able to map deformations of the skin and articular cartilage specimens to high resolution, in the order of 50 µm. This system can also be potentially used for the assessment of other biological tissues, bioengineered tissues or biomaterials with fine structures.

  12. Influence of the Drilling Mud Formulation Process on the Bacterial Communities in Thermogenic Natural Gas Wells of the Barnett Shale▿†

    Science.gov (United States)

    Struchtemeyer, Christopher G.; Davis, James P.; Elshahed, Mostafa S.

    2011-01-01

    The Barnett Shale in north central Texas contains natural gas generated by high temperatures (120 to 150°C) during the Mississippian Period (300 to 350 million years ago). In spite of the thermogenic origin of this gas, biogenic sulfide production and microbiologically induced corrosion have been observed at several natural gas wells in this formation. It was hypothesized that microorganisms in drilling muds were responsible for these deleterious effects. Here we collected drilling water and drilling mud samples from seven wells in the Barnett Shale during the drilling process. Using quantitative real-time PCR and microbial enumerations, we show that the addition of mud components to drilling water increased total bacterial numbers, as well as the numbers of culturable aerobic heterotrophs, acid producers, and sulfate reducers. The addition of sterile drilling muds to microcosms that contained drilling water stimulated sulfide production. Pyrosequencing-based phylogenetic surveys of the microbial communities in drilling waters and drilling muds showed a marked transition from typical freshwater communities to less diverse communities dominated by Firmicutes and Gammaproteobacteria. The community shifts observed reflected changes in temperature, pH, oxygen availability, and concentrations of sulfate, sulfonate, and carbon additives associated with the mud formulation process. Finally, several of the phylotypes observed in drilling muds belonged to lineages that were thought to be indigenous to marine and terrestrial fossil fuel formations. Our results suggest a possible alternative exogenous origin of such phylotypes via enrichment and introduction to oil and natural gas reservoirs during the drilling process. PMID:21602366

  13. High Dietary Fructose: Direct or Indirect Dangerous Factors Disturbing Tissue and Organ Functions

    Directory of Open Access Journals (Sweden)

    Dong-Mei Zhang

    2017-03-01

    Full Text Available High dietary fructose is a major contributor to insulin resistance and metabolic syndrome, disturbing tissue and organ functions. Fructose is mainly absorbed into systemic circulation by glucose transporter 2 (GLUT2 and GLUT5, and metabolized in liver to produce glucose, lactate, triglyceride (TG, free fatty acid (FFA, uric acid (UA and methylglyoxal (MG. Its extrahepatic absorption and metabolism also take place. High levels of these metabolites are the direct dangerous factors. During fructose metabolism, ATP depletion occurs and induces oxidative stress and inflammatory response, disturbing functions of local tissues and organs to overproduce inflammatory cytokine, adiponectin, leptin and endotoxin, which act as indirect dangerous factors. Fructose and its metabolites directly and/or indirectly cause oxidative stress, chronic inflammation, endothelial dysfunction, autophagy and increased intestinal permeability, and then further aggravate the metabolic syndrome with tissue and organ dysfunctions. Therefore, this review addresses fructose-induced metabolic syndrome, and the disturbance effects of direct and/or indirect dangerous factors on the functions of liver, adipose, pancreas islet, skeletal muscle, kidney, heart, brain and small intestine. It is important to find the potential correlations between direct and/or indirect risk factors and healthy problems under excess dietary fructose consumption.

  14. Fusing visual and clinical information for lung tissue classification in high-resolution computed tomography.

    Science.gov (United States)

    Depeursinge, Adrien; Racoceanu, Daniel; Iavindrasana, Jimison; Cohen, Gilles; Platon, Alexandra; Poletti, Pierre-Alexandre; Müller, Henning

    2010-09-01

    We investigate the influence of the clinical context of high-resolution computed tomography (HRCT) images of the chest on tissue classification. 2D regions of interest in HRCT axial slices from patients affected with an interstitial lung disease are automatically classified into five classes of lung tissue. Relevance of the clinical parameters is studied before fusing them with visual attributes. Two multimedia fusion techniques are compared: early versus late fusion. Early fusion concatenates features in one single vector, yielding a true multimedia feature space. Late fusion consisting of the combination of the probability outputs of two support vector machines. The late fusion scheme allowed a maximum of 84% correct predictions of testing instances among the five classes of lung tissue. This represents a significant improvement of 10% compared to a pure visual-based classification. Moreover, the late fusion scheme showed high robustness to the number of clinical parameters used, which suggests that it is appropriate for mining clinical attributes with missing values in clinical routine. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  15. High-contrast en bloc staining of neuronal tissue for field emission scanning electron microscopy.

    Science.gov (United States)

    Tapia, Juan Carlos; Kasthuri, Narayanan; Hayworth, Kenneth J; Schalek, Richard; Lichtman, Jeff W; Smith, Stephen J; Buchanan, JoAnn

    2012-01-12

    Conventional heavy metal poststaining methods on thin sections lend contrast but often cause contamination. To avoid this problem, we tested several en bloc staining techniques to contrast tissue in serial sections mounted on solid substrates for examination by field emission scanning electron microscopy (FESEM). Because FESEM section imaging requires that specimens have higher contrast and greater electrical conductivity than transmission electron microscopy (TEM) samples, our technique uses osmium impregnation (OTO) to make the samples conductive while heavily staining membranes for segmentation studies. Combining this step with other classic heavy metal en bloc stains, including uranyl acetate (UA), lead aspartate, copper sulfate and lead citrate, produced clean, highly contrasted TEM and scanning electron microscopy (SEM) samples of insect, fish and mammalian nervous systems. This protocol takes 7-15 d to prepare resin-embedded tissue, cut sections and produce serial section images.

  16. Use of flow cytometry for high-throughput cell population estimates in brain tissue

    Science.gov (United States)

    Young, Nicole A.; Flaherty, David K.; Airey, David C.; Varlan, Peter; Aworunse, Feyi; Kaas, Jon H.; Collins, Christine E.

    2012-01-01

    The large size of primate brains is an impediment to obtaining high-resolution cell number maps of the cortex in humans and non-human primates. We present a rapid, flow cytometry-based cell counting method that can be used to estimate cell numbers from homogenized brain tissue samples comprising the entire cortical sheet. The new method, called the flow fractionator, is based on the isotropic fractionator (IF) method (Herculano-Houzel and Lent, 2005), but substitutes flow cytometry analysis for manual, microscope analysis using a Neubauer counting chamber. We show that our flow cytometry-based method for total cell estimation in homogenized brain tissue provides comparable data to that obtained using a counting chamber on a microscope. The advantages of the flow fractionator over existing methods are improved precision of cell number estimates and improved speed of analysis. PMID:22798947

  17. High-definition Fourier transform infrared spectroscopic imaging of prostate tissue

    Science.gov (United States)

    Wrobel, Tomasz P.; Kwak, Jin Tae; Kadjacsy-Balla, Andre; Bhargava, Rohit

    2016-03-01

    Histopathology forms the gold standard for cancer diagnosis and therapy, and generally relies on manual examination of microscopic structural morphology within tissue. Fourier-Transform Infrared (FT-IR) imaging is an emerging vibrational spectroscopic imaging technique, especially in a High-Definition (HD) format, that provides the spatial specificity of microscopy at magnifications used in diagnostic surgical pathology. While it has been shown for standard imaging that IR absorption by tissue creates a strong signal where the spectrum at each pixel is a quantitative "fingerprint" of the molecular composition of the sample, here we show that this fingerprint also enables direct digital pathology without the need for stains or dyes for HD imaging. An assessment of the potential of HD imaging to improve diagnostic pathology accuracy is presented.

  18. High rates of detection of respiratory viruses in tonsillar tissues from children with chronic adenotonsillar disease.

    Directory of Open Access Journals (Sweden)

    Jose Luiz Proenca-Modena

    Full Text Available Chronic tonsillar diseases are an important health problem, leading to large numbers of surgical procedures worldwide. Little is known about pathogenesis of these diseases. In order to investigate the role of respiratory viruses in chronic adenotonsillar diseases, we developed a cross-sectional study to determine the rates of viral detections of common respiratory viruses detected by TaqMan real time PCR (qPCR in nasopharyngeal secretions, tonsillar tissues and peripheral blood from 121 children with chronic tonsillar diseases, without symptoms of acute respiratory infections. At least one respiratory virus was detected in 97.5% of patients. The viral co-infection rate was 69.5%. The most frequently detected viruses were human adenovirus in 47.1%, human enterovirus in 40.5%, human rhinovirus in 38%, human bocavirus in 29.8%, human metapneumovirus in 17.4% and human respiratory syncytial virus in 15.7%. Results of qPCR varied widely between sample sites: human adenovirus, human bocavirus and human enterovirus were predominantly detected in tissues, while human rhinovirus was more frequently detected in secretions. Rates of virus detection were remarkably high in tonsil tissues: over 85% in adenoids and close to 70% in palatine tonsils. In addition, overall virus detection rates were higher in more hypertrophic than in smaller adenoids (p = 0.05, and in the particular case of human enteroviruses, they were detected more frequently (p = 0.05 in larger palatine tonsils than in smaller ones. While persistence/latency of DNA viruses in tonsillar tissues has been documented, such is not the case of RNA viruses. Respiratory viruses are highly prevalent in adenoids and palatine tonsils of patients with chronic tonsillar diseases, and persistence of these viruses in tonsils may stimulate chronic inflammation and play a role in the pathogenesis of these diseases.

  19. High rates of detection of respiratory viruses in tonsillar tissues from children with chronic adenotonsillar disease.

    Science.gov (United States)

    Proenca-Modena, Jose Luiz; Pereira Valera, Fabiana Cardoso; Jacob, Marcos Gerhardinger; Buzatto, Guilherme Pietrucci; Saturno, Tamara Honorato; Lopes, Lucia; Souza, Jamila Mendonça; Escremim Paula, Flavia; Silva, Maria Lucia; Carenzi, Lucas Rodrigues; Tamashiro, Edwin; Arruda, Eurico; Anselmo-Lima, Wilma Terezinha

    2012-01-01

    Chronic tonsillar diseases are an important health problem, leading to large numbers of surgical procedures worldwide. Little is known about pathogenesis of these diseases. In order to investigate the role of respiratory viruses in chronic adenotonsillar diseases, we developed a cross-sectional study to determine the rates of viral detections of common respiratory viruses detected by TaqMan real time PCR (qPCR) in nasopharyngeal secretions, tonsillar tissues and peripheral blood from 121 children with chronic tonsillar diseases, without symptoms of acute respiratory infections. At least one respiratory virus was detected in 97.5% of patients. The viral co-infection rate was 69.5%. The most frequently detected viruses were human adenovirus in 47.1%, human enterovirus in 40.5%, human rhinovirus in 38%, human bocavirus in 29.8%, human metapneumovirus in 17.4% and human respiratory syncytial virus in 15.7%. Results of qPCR varied widely between sample sites: human adenovirus, human bocavirus and human enterovirus were predominantly detected in tissues, while human rhinovirus was more frequently detected in secretions. Rates of virus detection were remarkably high in tonsil tissues: over 85% in adenoids and close to 70% in palatine tonsils. In addition, overall virus detection rates were higher in more hypertrophic than in smaller adenoids (p = 0.05), and in the particular case of human enteroviruses, they were detected more frequently (p = 0.05) in larger palatine tonsils than in smaller ones. While persistence/latency of DNA viruses in tonsillar tissues has been documented, such is not the case of RNA viruses. Respiratory viruses are highly prevalent in adenoids and palatine tonsils of patients with chronic tonsillar diseases, and persistence of these viruses in tonsils may stimulate chronic inflammation and play a role in the pathogenesis of these diseases.

  20. A highly organized three-dimensional alginate scaffold for cartilage tissue engineering prepared by microfluidic technology.

    Science.gov (United States)

    Wang, Chen-Chie; Yang, Kai-Chiang; Lin, Keng-Hui; Liu, Hwa-Chang; Lin, Feng-Huei

    2011-10-01

    Osteoarthritis is a degenerative disease and frequently involves the knee, hip and phalangeal joints. Current treatments used in small cartilage defects including multiple drilling, abrasion arthroplasty, mosaicplasty, and autogenous chondrocyte transplantation, however, there are problems needed to be solved. The standard treatment for severe osteoarthritis is total joint arthroplasty. The disadvantages of this surgery are the possibility of implant loosening. Therefore, tissue engineering for cartilage regeneration has become a promising topic. We have developed a new method to produce a highly organized single polymer (alginate) scaffold using microfluidic device. Scanning electron microscope and confocal fluoroscope examinations showed that the scaffold has a regular interconnected porous structure in the scale of 250 μm and high porosity. The scaffold is effective in chondrocyte culture; the cell viability test (WST-1 assay), cell toxicity (lactate dehydrogenase assay), cell survival rate, extracellular matrix production (glycosaminoglycans contents), cell proliferation (DNA quantification), and gene expression (real-time PCR) all revealed good results for chondrocyte culture. The chondrocytes can maintain normal phenotypes, highly express aggrecan and type II collagen, and secrete a great deal of extracellular matrix when seeded in the alginate scaffold. This study demonstrated that a highly organized alginate scaffold can be prepared with an economical microfluidic device, and this scaffold is effective in cartilage tissue engineering. Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.

  1. High levels of DNA polymerase β mRNA corresponding with the high activity in Graves' thyroid tissue.

    Science.gov (United States)

    Hayakawa, N; Sato, Y; Nagasaka, A; Mano, Y; Nagasaka, T; Nakai, A; Iwase, K; Yoshida, S

    2017-04-01

    High DNA polymerase β activity has been observed in the thyroid tissue of patients with Graves' disease (Nagasaka et al. in Metabolism 37:1051-1054, 1988). This fact aroused our interest in whether the alteration of DNA polymerase β activity depends on DNA polymerase β (DNA poly β) mRNA levels, which may be modulated by thyroid-stimulating hormone (TSH) or thyroid-stimulating substances, i.e. TSH receptor antibody (TRAb). Addition of TSH or TRAb to primary cultures of Graves' disease thyroid cells for 4 h led to no increase in DNA poly β mRNA levels. In contrast, thyroid hormone synthesizing enzyme, peroxidase, mRNA levels increased fivefold after coculture with TSH and TRAb, even though DNA poly β activity and mRNA levels are already significantly higher in Graves' disease thyroid tissues, compared with normal thyroid tissue. These results indicate that DNA poly β expression in Graves' disease thyroid cells may be maximally activated or plateau in response to thyroid-stimulating immunoglobulins, or that the activation of to poly β expression may occur via pathways other than the G protein and cyclic AMP system.

  2. High-fat diet leads to tissue-specific changes reflecting risk factors for diseases in DBA/2J mice

    Science.gov (United States)

    Hageman, Rachael S.; Wagener, Asja; Hantschel, Claudia; Svenson, Karen L.; Churchill, Gary A.

    2010-01-01

    The aim of this study was to characterize the responses of individual tissues to high-fat feeding as a function of mass, fat composition, and transcript abundance. We examined a panel of eight tissues [5 white adipose tissues (WAT), brown adipose tissue (BAT), liver, muscle] obtained from DBA/2J mice on either a standard breeding diet (SBD) or a high-fat diet (HFD). HFD led to weight gain, decreased insulin sensitivity, and tissue-specific responses, including inflammation, in these mice. The dietary fatty acids were partially metabolized and converted in both liver and fat tissues. Saturated fatty acids (SFA) were converted in the liver to monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA), and oleic acid (C18:1) was the preferred MUFA for storage of excess energy in all tissues of HFD-fed mice. Transcriptional changes largely reflected the tissue-specific fat deposition. SFA were negatively correlated with genes in the collagen family and processes involving the extracellular matrix. We propose a novel role of the tryptophan hydroxylase 2 (Tph2) gene in adipose tissues of diet-induced obesity. Tissue-specific responses to HFD were identified. Liver steatosis was evident in HFD-fed mice. Gonadal, retroperitoneal and subcutaneous adipose tissue and BAT exhibited severe inflammatory and immune responses. Mesenteric adipose tissue was the most metabolically active adipose tissue. Gluteal adipose tissue had the highest mass gain but was sluggish in its metabolism. In HFD conditions, BAT functioned largely like WAT in its role as a depot for excess energy, whereas WAT played a role in thermogenesis. PMID:20215417

  3. An UPLC-MS/MS method for highly sensitive high-throughput analysis of phytohormones in plant tissues

    Directory of Open Access Journals (Sweden)

    Balcke Gerd Ulrich

    2012-11-01

    Full Text Available Abstract Background Phytohormones are the key metabolites participating in the regulation of multiple functions of plant organism. Among them, jasmonates, as well as abscisic and salicylic acids are responsible for triggering and modulating plant reactions targeted against pathogens and herbivores, as well as resistance to abiotic stress (drought, UV-irradiation and mechanical wounding. These factors induce dramatic changes in phytohormone biosynthesis and transport leading to rapid local and systemic stress responses. Understanding of underlying mechanisms is of principle interest for scientists working in various areas of plant biology. However, highly sensitive, precise and high-throughput methods for quantification of these phytohormones in small samples of plant tissues are still missing. Results Here we present an LC-MS/MS method for fast and highly sensitive determination of jasmonates, abscisic and salicylic acids. A single-step sample preparation procedure based on mixed-mode solid phase extraction was efficiently combined with essential improvements in mobile phase composition yielding higher efficiency of chromatographic separation and MS-sensitivity. This strategy resulted in dramatic increase in overall sensitivity, allowing successful determination of phytohormones in small (less than 50 mg of fresh weight tissue samples. The method was completely validated in terms of analyte recovery, sensitivity, linearity and precision. Additionally, it was cross-validated with a well-established GC-MS-based procedure and its applicability to a variety of plant species and organs was verified. Conclusion The method can be applied for the analyses of target phytohormones in small tissue samples obtained from any plant species and/or plant part relying on any commercially available (even less sensitive tandem mass spectrometry instrumentation.

  4. An UPLC-MS/MS method for highly sensitive high-throughput analysis of phytohormones in plant tissues

    Science.gov (United States)

    2012-01-01

    Background Phytohormones are the key metabolites participating in the regulation of multiple functions of plant organism. Among them, jasmonates, as well as abscisic and salicylic acids are responsible for triggering and modulating plant reactions targeted against pathogens and herbivores, as well as resistance to abiotic stress (drought, UV-irradiation and mechanical wounding). These factors induce dramatic changes in phytohormone biosynthesis and transport leading to rapid local and systemic stress responses. Understanding of underlying mechanisms is of principle interest for scientists working in various areas of plant biology. However, highly sensitive, precise and high-throughput methods for quantification of these phytohormones in small samples of plant tissues are still missing. Results Here we present an LC-MS/MS method for fast and highly sensitive determination of jasmonates, abscisic and salicylic acids. A single-step sample preparation procedure based on mixed-mode solid phase extraction was efficiently combined with essential improvements in mobile phase composition yielding higher efficiency of chromatographic separation and MS-sensitivity. This strategy resulted in dramatic increase in overall sensitivity, allowing successful determination of phytohormones in small (less than 50 mg of fresh weight) tissue samples. The method was completely validated in terms of analyte recovery, sensitivity, linearity and precision. Additionally, it was cross-validated with a well-established GC-MS-based procedure and its applicability to a variety of plant species and organs was verified. Conclusion The method can be applied for the analyses of target phytohormones in small tissue samples obtained from any plant species and/or plant part relying on any commercially available (even less sensitive) tandem mass spectrometry instrumentation. PMID:23173950

  5. Role of the autonomic nervous system in the thermogenic response to food in lean individuals.

    Science.gov (United States)

    De Jonge, L; Garrel, D R

    1997-05-01

    The aim of this study was to determine the role of the autonomic nervous system (ANS) in obligatory and facultative components of the thermogenic response to food (TRF). Nineteen lean, healthy subjects participated in this study, which comprised two protocols, each exploring one component of the ANS. In the first experimental group, propranolol (prime: 80 micrograms/kg; continuous: 1 microgram.kg-1.min-1) was infused intravenously to inhibit sympathetic nervous activity (SNA), whereas in the second group atropine (prime: 5 micrograms/kg; continuous: 5 micrograms.kg-1.min-1) was used to inhibit parasympathetic nervous activity (PNA). The TRF was measured on four occasions: 1) after oral ingestion of a breakfast, during 0.9% NaCl perfusion, 2) after oral ingestion of the same breakfast, during the perfusion of one of the drugs, 3) after intragastric injection of a pureed form of the same meal as in part 1, during 0.9% NaCl perfusion, and 4) after intragastric feeding, during the administration of one of the drugs. Energy expenditure was measured by indirect calorimetry for 30 min before and 6 h after ingestion of the meal. Facultative TRF was defined as the difference between oral and intragastric TRF. Intragastric feeding significantly reduced TRF in both studies: 6.6 +/- 1.0 vs. 8.7 +/- 0.8% of the ingested energy in the SNA study and 5.5 +/- 1.6 vs. 7.4 +/- 3.1% in the PNA study. During propranolol infusion, TRF was significantly lower than it was during saline infusion after oral feeding (6.9 +/- 1.0% vs. 8.7 +/- 0.8% of ingested energy) but not after intragastric feeding. During atropine administration, TRF was reduced after both oral and intragastric feeding, although statistical significance was not reached in the latter. Atropine administration decreased gastric emptying (measured with an isotopic method) 2 h postingestion by 50%. These results show that the SNA is necessary for the facultative component of TRF to occur in humans. The role of the PNA appears

  6. Central regulation of brown adipose tissue thermogenesis and energy homeostasis dependent on food availability.

    Science.gov (United States)

    Nakamura, Yoshiko; Nakamura, Kazuhiro

    2017-12-05

    Energy homeostasis of mammals is maintained by balancing energy expenditure within the body and energy intake through feeding. Several lines of evidence indicate that brown adipose tissue (BAT), a sympathetically activated thermogenic organ, turns excess energy into heat to maintain the energy balance in rodents and humans, in addition to its thermoregulatory role for the defense of body core temperature in cold environments. Elucidating the central circuit mechanism controlling BAT thermogenesis dependent on nutritional conditions and food availability in relation to energy homeostasis is essential to understand the etiology of symptoms caused by energy imbalance, such as obesity. The central thermogenic command outflow to BAT descends through an excitatory neural pathway mediated by hypothalamic, medullary and spinal sites. This sympathoexcitatory thermogenic drive is controlled by tonic GABAergic inhibitory signaling from the thermoregulatory center in the preoptic area, whose tone is altered by body core and cutaneous thermosensory inputs. This circuit controlling BAT thermogenesis for cold defense also functions for the development of fever and psychological stress-induced hyperthermia, indicating its important role in the defense from a variety of environmental stressors. When food is unavailable, hunger-driven neural signaling from the hypothalamus activates GABAergic neurons in the medullary reticular formation, which then block the sympathoexcitatory thermogenic outflow to BAT to reduce energy expenditure and simultaneously command the masticatory motor system to promote food intake-effectively commanding responses to survive starvation. This article reviews the central mechanism controlling BAT thermogenesis in relation to the regulation of energy and thermal homeostasis dependent on food availability.

  7. Centimeter-deep tissue fluorescence microscopic imaging with high signal-to-noise ratio and picomole sensitivity

    CERN Document Server

    Cheng, Bingbing; Wei, Ming-Yuan; Pei, Yanbo; DSouza, Francis; Nguyen, Kytai T; Hong, Yi; Tang, Liping; Yuan, Baohong

    2015-01-01

    Fluorescence microscopic imaging in centimeter-deep tissue has been highly sought-after for many years because much interesting in vivo micro-information, such as microcirculation, tumor angiogenesis, and metastasis, may deeply locate in tissue. In this study, for the first time this goal has been achieved in 3-centimeter deep tissue with high signal-to-noise ratio (SNR) and picomole sensitivity under radiation safety thresholds. These results are demonstrated not only in tissue-mimic phantoms but also in actual tissues, such as porcine muscle, ex vivo mouse liver, ex vivo spleen, and in vivo mouse tissue. These results are achieved based on three unique technologies: excellent near infrared ultrasound-switchable fluorescence (USF) contrast agents, a sensitive USF imaging system, and an effective correlation method. Multiplex USF fluorescence imaging is also achieved. It is useful to simultaneously image multiple targets and observe their interactions. This work opens the door for future studies of centimeter...

  8. High Resolution Magic Angle Spinning 1H-NMR Metabolic Profiling of Nanoliter Biological Tissues at High Magnetic Field

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Ju; Hu, Jian Z.; Burton, Sarah D.; Hoyt, David W.

    2013-03-05

    It is demonstrated that a high resolution magic angle spinning 1H-NMR spectrum of biological tissue samples with volumes as small as 150 nanoliters, or 0.15 mg in weight, can be acquired in a few minutes at 21.1 T magnetic field using a commercial 1.6 mm fast-MAS probe with minor modification of the MAS rotor. The strategies of sealing the samples inside the MAS rotor to avoid fluid leakage as well as the ways of optimizing the signal to noise are discussed.

  9. Preparation and characterization of highly porous, biodegradable polyurethane scaffolds for soft tissue applications

    Science.gov (United States)

    Guan, Jianjun; Fujimoto, Kazuro L.; Sacks, Michael S.; Wagner, William R.

    2010-01-01

    In the engineering of soft tissues, scaffolds with high elastance and strength coupled with controllable biodegradable properties are necessary. To fulfill such design criteria we have previously synthesized two kinds of biodegradable polyurethaneureas, namely poly(ester urethane)urea (PEUU) and poly(ether ester urethane)urea (PEEUU) from polycaprolactone, polycaprolactone-b-polyethylene glycol-b-polycaprolactone, 1,4-diisocyanatobutane and putrescine. PEUU and PEEUU were further fabricated into scaffolds by thermally induced phase separation using dimethyl sulfoxide (DMSO) as a solvent. The effect of polymer solution concentration, quenching temperature and polymer type on pore morphology and porosity was investigated. Scaffolds were obtained with open and interconnected pores having sizes ranging from several μm to more than 150 μm and porosities of 80–97%. By changing the polymer solution concentration or quenching temperature, scaffolds with random or oriented tubular pores could be obtained. The PEUU scaffolds were flexible with breaking strains of 214% and higher, and tensile strengths of approximately 1.0 MPa, whereas the PEEUU scaffolds generally had lower strengths and breaking strains. Scaffold degradation in aqueous buffer was related to the porosity and polymer hydrophilicity. Smooth muscle cells were filtration seeded in the scaffolds and it was shown that both scaffolds supported cell adhesion and growth, with smooth muscle cells growing more extensively in the PEEUU scaffold. These biodegradable and flexible scaffolds demonstrate potential for future application as cell scaffolds in cardiovascular tissue engineering or other soft tissue applications. PMID:15626443

  10. Adipose Tissue CLK2 Promotes Energy Expenditure during High-Fat Diet Intermittent Fasting.

    Science.gov (United States)

    Hatting, Maximilian; Rines, Amy K; Luo, Chi; Tabata, Mitsuhisa; Sharabi, Kfir; Hall, Jessica A; Verdeguer, Francisco; Trautwein, Christian; Puigserver, Pere

    2017-02-07

    A promising approach to treating obesity is to increase diet-induced thermogenesis in brown adipose tissue (BAT), but the regulation of this process remains unclear. Here we find that CDC-like kinase 2 (CLK2) is expressed in BAT and upregulated upon refeeding. Mice lacking CLK2 in adipose tissue exhibit exacerbated obesity and decreased energy expenditure during high-fat diet intermittent fasting. Additionally, tissue oxygen consumption and protein levels of UCP1 are reduced in CLK2-deficient BAT. Phosphorylation of CREB, a transcriptional activator of UCP1, is markedly decreased in BAT cells lacking CLK2 due to enhanced CREB dephosphorylation. Mechanistically, CREB dephosphorylation is rescued by the inhibition of PP2A, a phosphatase that targets CREB. Our results suggest that CLK2 is a regulatory component of diet-induced thermogenesis in BAT through increased CREB-dependent expression of UCP1. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Extraction of high-quality DNA from ethanol-preserved tropical plant tissues.

    Science.gov (United States)

    Bressan, Eduardo A; Rossi, Mônica L; Gerald, Lee T S; Figueira, Antonio

    2014-04-24

    Proper conservation of plant samples, especially during remote field collection, is essential to assure quality of extracted DNA. Tropical plant species contain considerable amounts of secondary compounds, such as polysaccharides, phenols, and latex, which affect DNA quality during extraction. The suitability of ethanol (96% v/v) as a preservative solution prior to DNA extraction was evaluated using leaves of Jatropha curcas and other tropical species. Total DNA extracted from leaf samples stored in liquid nitrogen or ethanol from J. curcas and other tropical species (Theobroma cacao, Coffea arabica, Ricinus communis, Saccharum spp., and Solanum lycopersicon) was similar in quality, with high-molecular-weight DNA visualized by gel electrophoresis. DNA quality was confirmed by digestion with EcoRI or HindIII and by amplification of the ribosomal gene internal transcribed spacer region. Leaf tissue of J. curcas was analyzed by light and transmission electron microscopy before and after exposure to ethanol. Our results indicate that leaf samples can be successfully preserved in ethanol for long periods (30 days) as a viable method for fixation and conservation of DNA from leaves. The success of this technique is likely due to reduction or inactivation of secondary metabolites that could contaminate or degrade genomic DNA. Tissue conservation in 96% ethanol represents an attractive low-cost alternative to commonly used methods for preservation of samples for DNA extraction. This technique yields DNA of equivalent quality to that obtained from fresh or frozen tissue.

  12. Synthesis of highly interconnected 3D scaffold from Arothron stellatus skin collagen for tissue engineering application.

    Science.gov (United States)

    Ramanathan, Giriprasath; Singaravelu, Sivakumar; Raja, M D; Sivagnanam, Uma Tiruchirapalli

    2015-11-01

    The substrate which is avidly used for tissue engineering applications should have good mechanical and biocompatible properties, and all these parameters are often considered as essential for dermal reformation. Highly interconnected three dimensional (3D) wound dressing material with enhanced structural integrity was synthesized from Arothron stellatus fish skin (AsFS) collagen for tissue engineering applications. The synthesized 3D collagen sponge (COL-SPG) was further characterized by different physicochemical methods. The scanning electron microscopy analysis of the material demonstrated that well interconnected pores with homogeneous microstructure on the surface aids higher swelling index and that the material also possessed good mechanical properties with a Young's modulus of 0.89±0.2 MPa. Biocompatibility of the 3D COL-SPG showed 92% growth for both NIH 3T3 fibroblasts and keratinocytes. Overall, the study revealed that synthesized 3D COL-SPG from fish skin will act as a promising wound dressing in skin tissue engineering. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Highly specific sites of prolactin binding in benign and malignant breast tissue.

    Science.gov (United States)

    Agarwal, P K; Tandon, S; Agrawal, A K; Kumar, S

    1989-12-01

    An immunocytochemical method involving the application of polyclonal antisera to human prolactin (PRL) followed by a highly sensitive and a modified version of dinitrophenyl (DNP) hapten sandwich staining procedure using anti-DNP IgM monoclonal antibody has been used to detect PRL binding in benign and malignant breast tissue. The technique was applied to 5 microns thick sections of paraffin embedded formalin fixed tissue. Out of 107 breast biopsies 40 were carcinomas, 41 were fibroadenomas, 18 were benign cystic disease and 8 were gynaecomastia. In cases of carcinoma positive staining was observed in 82.5% cases whereas in fibroadenoma the positivity was in 57% cases only. The positive reaction in fibroadenoma was mainly due to the presence of apocrine metaplasia associated with the tumour. Also PRL was present in greater proportion in post menopausal patients as compared to premenopausal cancer patients. These findings suggest the presence of specific PRL binding sites in breast tissue. The staining was restricted to epithelial cells and background staining of the stroma was minimally seen in these cases. Positively stained breast carcinoma may represent an apocrine subset of the carcinoma.

  14. Experimental high-intensity focused ultrasound lesion formation in cardiac tissue

    Science.gov (United States)

    Muratore, Robert; Kalisz, Andrew; Lee, Paul; Lizzi, Frederic; Fujikura, Kana; Otsuka, Ryo; Homma, Shunichi

    2004-05-01

    High-intensity focused ultrasound (HIFU) (4.5-7.5 MHz) was used to form lesions in cardiac tissue, with an ultimate objective of treating conditions such as hypertrophic cardiomyopathy and ventricular tachycardia. Ultrasound attenuation coefficients were experimentally determined in vitro for calf myocardial tissue, both muscle and pericardial fat. These coefficients were employed in computational models of linear beam propagation, tissue heating profiles and thermal lesion formation for a variety of focused transducers. Modeling was performed for continuous and pulsed exposures. These models suggested initial power levels and exposure durations for in vitro experiments on calf ventricles and septa and ex vivo experiments on canine whole hearts. Repeatability of lesion size and placement was studied as power and exposure parameters varied around the initial values. With these experimental results, power and exposure parameters were selected to create lesions in vivo in canine ventricles and septa in open-chest, anesthetized dogs. Pulsed exposures were synchronized to cardiac and respiration cycles to ensure accurate placement of the lesions. These initial in vivo experiments showed that HIFU treatments in the beating heart are feasible; they also identified refinements that are now being implemented for better control of lesion size and placement. [Work supported by NCI and NHLBI Grant 5R01 CA84588.

  15. Advanced CLARITY for rapid and high-resolution imaging of intact tissues.

    Science.gov (United States)

    Tomer, Raju; Ye, Li; Hsueh, Brian; Deisseroth, Karl

    2014-07-01

    CLARITY is a method for chemical transformation of intact biological tissues into a hydrogel-tissue hybrid, which becomes amenable to interrogation with light and macromolecular labels while retaining fine structure and native biological molecules. This emerging accessibility of information from large intact samples has created both new opportunities and new challenges. Here we describe protocols spanning multiple dimensions of the CLARITY workflow, ranging from simple, reliable and efficient lipid removal without electrophoretic instrumentation (passive CLARITY) to optimized objectives and integration with light-sheet optics (CLARITY-optimized light-sheet microscopy (COLM)) for accelerating data collection from clarified samples by several orders of magnitude while maintaining or increasing quality and resolution. The entire protocol takes from 7-28 d to complete for an adult mouse brain, including hydrogel embedding, full lipid removal, whole-brain antibody staining (which, if needed, accounts for 7-10 of the days), and whole-brain high-resolution imaging; timing within this window depends on the choice of lipid removal options, on the size of the tissue, and on the number and type of immunostaining rounds performed. This protocol has been successfully applied to the study of adult mouse, adult zebrafish and adult human brains, and it may find many other applications in the structural and molecular analysis of large assembled biological systems.

  16. A high-resolution model for soft tissue deformation based on point primitives.

    Science.gov (United States)

    Zou, Yanni; Liu, Peter X

    2017-09-01

    In order to achieve a high degree of visual realism in surgery simulation, we propose a new model, which is based on point primitives and continuous elastic mechanics theory, for soft tissue deformation, tearing and/or cutting. The model can be described as a two-step local high-resolution strategy. First, appropriate volumetric data are sampled and assigned with proper physical properties. Second, sparsely sampled points in non-deformed regions and densely-sampled points in the deformed zone are selected and evaluated. By using a meshless deformation model based on point primitives for all volumetric data, the affine transform matrix of collision points can be computed. The new positions of neighboring points in the collided surface can be then calculated, and more details in the local deformed zone can be obtained for rendering. Technical details about the derivations of the proposed model as well as its implementation are given. The visual effects and computation cost of the proposed model are evaluated and compared with conventional primitives-based methods. Experimental results show that the proposed model provides users (trainees) with improved visual feedback while the computational cost is at the same magnitude of other similar methods. The proposed method is especially suitable for the simulation of soft tissue deformation and tearing because no grid information needs to be maintained. It can simulate soft tissue deformation in a high degree of authenticity with real-time performance. It could be considered implemented in the development of a mixed reality application of neurosurgery simulators in the future. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. An in vitro scratch tendon tissue injury model: effects of high frequency low magnitude loading.

    Science.gov (United States)

    Adekanmbi, Isaiah; Zargar, Nasim; Hulley, Philippa

    2017-03-01

    The healing process of ruptured tendons is suboptimal, taking months to achieve tissue with inferior properties to healthy tendon. Mechanical loading has been shown to positively influence tendon healing. However, high frequency low magnitude (HFLM) loads, which have shown promise in maintaining healthy tendon properties, have not been studied with in vitro injury models. Here, we present and validate an in vitro scratch tendon tissue injury model to investigate effects of HFLM loading on the properties of injured rat tail tendon fascicles (RTTFs). A longitudinal tendon tear was simulated using a needle aseptically to scratch a defined length along individual RTTFs. Tissue viability, biomechanical, and biochemical parameters were investigated before and 7 days after culture . The effects of static, HFLM (20 Hz), and low frequency (1 Hz) cyclic loading or no load were also investigated. Tendon viability was confirmed in damaged RTTFs after 7 days of culture, and the effects of a 0.77 ± 0.06 cm scratch on the mechanical property (tangent modulus) and tissue metabolism in damaged tendons were consistent, showing significant damage severity compared with intact tendons. Damaged tendon fascicles receiving HFLM (20 Hz) loads displayed significantly higher mean tangent modulus than unloaded damaged tendons (212.7 ± 14.94 v 92.7 ± 15.59 MPa), and damaged tendons receiving static loading (117.9 ± 10.65 MPa). HFLM stimulation maintained metabolic activity in 7-day cultured damaged tendons at similar levels to fresh tendons immediately following damage. Only damaged tendons receiving HFLM loads showed significantly higher metabolism than unloaded damaged tendons (relative fluorescence units -7021 ± 635.9 v 3745.1 ± 641.7). These validation data support the use of the custom-made in vitro injury model for investigating the potential of HFLM loading interventions in treating damaged tendons.

  18. Feasibility analysis of high resolution tissue image registration using 3-D synthetic data.

    Science.gov (United States)

    Sharma, Yachna; Moffitt, Richard A; Stokes, Todd H; Chaudry, Qaiser; Wang, May D

    2011-01-01

    Registration of high-resolution tissue images is a critical step in the 3D analysis of protein expression. Because the distance between images (~4-5μm thickness of a tissue section) is nearly the size of the objects of interest (~10-20μm cancer cell nucleus), a given object is often not present in both of two adjacent images. Without consistent correspondence of objects between images, registration becomes a difficult task. This work assesses the feasibility of current registration techniques for such images. We generated high resolution synthetic 3-D image data sets emulating the constraints in real data. We applied multiple registration methods to the synthetic image data sets and assessed the registration performance of three techniques (i.e., mutual information (MI), kernel density estimate (KDE) method [1], and principal component analysis (PCA)) at various slice thicknesses (with increments of 1μm) in order to quantify the limitations of each method. Our analysis shows that PCA, when combined with the KDE method based on nuclei centers, aligns images corresponding to 5μm thick sections with acceptable accuracy. We also note that registration error increases rapidly with increasing distance between images, and that the choice of feature points which are conserved between slices improves performance. We used simulation to help select appropriate features and methods for image registration by estimating best-case-scenario errors for given data constraints in histological images. The results of this study suggest that much of the difficulty of stained tissue registration can be reduced to the problem of accurately identifying feature points, such as the center of nuclei.

  19. Feasibility analysis of high resolution tissue image registration using 3-D synthetic data

    Directory of Open Access Journals (Sweden)

    Yachna Sharma

    2011-01-01

    Full Text Available Background: Registration of high-resolution tissue images is a critical step in the 3D analysis of protein expression. Because the distance between images (~4-5μm thickness of a tissue section is nearly the size of the objects of interest (~10-20μm cancer cell nucleus, a given object is often not present in both of two adjacent images. Without consistent correspondence of objects between images, registration becomes a difficult task. This work assesses the feasibility of current registration techniques for such images. Methods: We generated high resolution synthetic 3-D image data sets emulating the constraints in real data. We applied multiple registration methods to the synthetic image data sets and assessed the registration performance of three techniques (i.e., mutual information (MI, kernel density estimate (KDE method [1], and principal component analysis (PCA at various slice thicknesses (with increments of 1μm in order to quantify the limitations of each method. Results: Our analysis shows that PCA, when combined with the KDE method based on nuclei centers, aligns images corresponding to 5μm thick sections with acceptable accuracy. We also note that registration error increases rapidly with increasing distance between images, and that the choice of feature points which are conserved between slices improves performance. Conclusions: We used simulation to help select appropriate features and methods for image registration by estimating best-case-scenario errors for given data constraints in histological images. The results of this study suggest that much of the difficulty of stained tissue registration can be reduced to the problem of accurately identifying feature points, such as the center of nuclei.

  20. Analyzing the Function of Cartilage Replacements: A Laboratory Activity to Teach High School Students Chemical and Tissue Engineering Concepts

    Science.gov (United States)

    Renner, Julie N.; Emady, Heather N.; Galas, Richards J., Jr.; Zhange, Rong; Baertsch, Chelsey D.; Liu, Julie C.

    2013-01-01

    A cartilage tissue engineering laboratory activity was developed as part of the Exciting Discoveries for Girls in Engineering (EDGE) Summer Camp sponsored by the Women In Engineering Program (WIEP) at Purdue University. Our goal was to increase awareness of chemical engineering and tissue engineering in female high school students through a…

  1. High expression of B7-H6 in human glioma tissues promotes tumor progression

    Science.gov (United States)

    Jiang, Tianwei; Wu, Wei; Zhang, Huasheng; Zhang, Xiangsheng; Zhang, Dingding; Wang, Qiang; Huang, Lei; Wang, Ye; Hang, Chunhua

    2017-01-01

    B7-H6, a new member of B7-family ligand, also known as NCR3LG1, plays an important role in NK cells mediated immune responses. Many studies have shown that it is highly expressed in various human cancers, and its expression levels are significantly associated with cancer patients’ clinicopathological parameters and postoperative prognoses. But, still the exact role of B7-H6 expression in human glioma remains elusive. In the present study, we have characterized the B7-H6 expression in the human glioma tissues as well as glioma cell lines, U87 and U251. We observed that B7-H6 was highly expressed in the human glioma tissues, and its expression was significantly associated with cancer progression. By using the RNA interference technology, we successfully ablated B7-H6 expression in human glioma cell lines to further study its contribution towards various biological features of this malignancy. Our study identified that the B7-H6 knockdown in U87 and U251 glioma cells significantly suppressed cell proliferation, migration, invasion, and enhanced apoptosis along with induction of cell cycle arrest. It thus suggested that B7-H6 play an important role in the regulation of the biological behavior of these glioma cells. However, the detailed mechanism of B7-H6 mediated regulation of glioma cancer cell transformation and its prognostic value merits further investigation. PMID:28415577

  2. Highly Concentrated Alginate-Gellan Gum Composites for 3D Plotting of Complex Tissue Engineering Scaffolds

    Directory of Open Access Journals (Sweden)

    Ashwini Rahul Akkineni

    2016-04-01

    Full Text Available In tissue engineering, additive manufacturing (AM technologies have brought considerable progress as they allow the fabrication of three-dimensional (3D structures with defined architecture. 3D plotting is a versatile, extrusion-based AM technology suitable for processing a wide range of biomaterials including hydrogels. In this study, composites of highly concentrated alginate and gellan gum were prepared in order to combine the excellent printing properties of alginate with the favorable gelling characteristics of gellan gum. Mixtures of 16.7 wt % alginate and 2 or 3 wt % gellan gum were found applicable for 3D plotting. Characterization of the resulting composite scaffolds revealed an increased stiffness in the wet state (15%–20% higher Young’s modulus and significantly lower volume swelling in cell culture medium compared to pure alginate scaffolds (~10% vs. ~23%. Cytocompatibility experiments with human mesenchymal stem cells (hMSC revealed that cell attachment was improved—the seeding efficiency was ~2.5–3.5 times higher on the composites than on pure alginate. Additionally, the composites were shown to support hMSC proliferation and early osteogenic differentiation. In conclusion, print fidelity of highly concentrated alginate-gellan gum composites was comparable to those of pure alginate; after plotting and crosslinking, the scaffolds possessed improved qualities regarding shape fidelity, mechanical strength, and initial cell attachment making them attractive for tissue engineering applications.

  3. Photoacoustic contrast imaging of biological tissues with nanodiamonds fabricated for high near-infrared absorbance.

    Science.gov (United States)

    Zhang, Ti; Cui, Huizhong; Fang, Chia-Yi; Su, Long-Jyun; Ren, Shenqiang; Chang, Huan-Cheng; Yang, Xinmai; Forrest, M Laird

    2013-02-01

    Radiation-damaged nanodiamonds (DNDs) are potentially ideal optical contrast agents for photoacoustic (PA) imaging in biological tissues due to their low toxicity and high optical absorbance. PA imaging contrast agents have been limited to quantum dots and gold particles, since most existing carbon-based nanoparticles, including fluorescent nanodiamonds, do not have sufficient optical absorption in the near-infrared (NIR) range. A new DND by He+ ion beam irradiation with very high NIR absorption was synthesized. These DNDs produced a 71-fold higher PA signal on a molar basis than similarly dimensioned gold nanorods, and 7.1 fmol of DNDs injected into rodents could be clearly imaged 3 mm below the skin surface with PA signal enhancement of 567% using an 820-nm laser wavelength.

  4. Phase contrast enhanced high resolution X-ray imaging and tomography of soft tissue

    Energy Technology Data Exchange (ETDEWEB)

    Jakubek, Jan [Institute of Experimental and Applied Physis, Czech Technical Universtiy in Prague, Horska 3a/22, 128 00 Prague 2 (Czech Republic); Granja, Carlos [Institute of Experimental and Applied Physis, Czech Technical Universtiy in Prague, Horska 3a/22, 128 00 Prague 2 (Czech Republic)]. E-mail: carlos.granja@utef.cvut.cz; Dammer, Jiri [Institute of Experimental and Applied Physis, Czech Technical Universtiy in Prague, Horska 3a/22, 128 00 Prague 2 (Czech Republic); Hanus, Robert [Institute of Organic Chemistry and Biochemistry, Academy of Sciences, CZ-166 10 Prague 6 (Czech Republic); Holy, Tomas [Institute of Experimental and Applied Physis, Czech Technical Universtiy in Prague, Horska 3a/22, 128 00 Prague 2 (Czech Republic); Pospisil, Stanislav [Institute of Experimental and Applied Physis, Czech Technical Universtiy in Prague, Horska 3a/22, 128 00 Prague 2 (Czech Republic); Tykva, Richard [Institute of Organic Chemistry and Biochemistry, Academy of Sciences, CZ-166 10 Prague 6 (Czech Republic); Uher, Josef [Institute of Experimental and Applied Physis, Czech Technical Universtiy in Prague, Horska 3a/22, 128 00 Prague 2 (Czech Republic); Vykydal, Zdenek [Institute of Experimental and Applied Physis, Czech Technical Universtiy in Prague, Horska 3a/22, 128 00 Prague 2 (Czech Republic)

    2007-02-01

    A tabletop system for digital high resolution and high sensitivity X-ray micro-radiography has been developed for small-animal and soft-tissue imaging. The system is based on a micro-focus X-ray tube and the semiconductor hybrid position sensitive Medipix2 pixel detector. Transmission radiography imaging, conventionally based only on absorption, is enhanced by exploiting phase-shift effects induced in the X-ray beam traversing the sample. Phase contrast imaging is realized by object edge enhancement. DAQ is done by a novel fully integrated USB-based readout with online image generation. Improved signal reconstruction techniques make use of advanced statistical data analysis, enhanced beam hardening correction and direct thickness calibration of individual pixels. 2D and 3D micro-tomography images of several biological samples demonstrate the applicability of the system for biological and medical purposes including in-vivo and time dependent physiological studies in the life sciences.

  5. Adjuvant high dose rate brachytherapy for soft tissue sarcomas: initial experience report

    Directory of Open Access Journals (Sweden)

    Robert Whitfield

    2011-03-01

    Full Text Available Purpose: Adjuvant high-dose-rate brachytherapy (HDRBT offers advantages over low dose rate brachytherapy (LDRBT, although there are little data on local tumor control and treatment related toxicity. We report outcome in patients with primary, recurrent, and metastatic extremity and superficial trunk soft tissue sarcoma. Material and methods: Eleven patients (12 sites with intermediate or high grade sarcoma were treated with adjuvant HDRBT following surgical resection. Patients were treated at 3.4 Gy fractions delivered twice daily to a total dose of 34 Gy (1 patient received 9 fractions. Results: With median follow-up of 20.8 months, 1 patient developed a local recurrence. 2-year local control and overall survival are 89% and 71%, respectively. Wound complications occurred in 3 sites. Two of the wound complications developed in the area of previous external beam radiotherapy (EBRT. Conclusion: Surgical resection followed by HDRBT is associated with excellent early local tumor control and acceptable wound complication.

  6. The effects of Magnetic Resonance Imaging-guided High-Intensity Focused Ultrasound ablation on human cadaver breast tissue.

    Science.gov (United States)

    Merckel, Laura G; Deckers, Roel; Baron, Paul; Bleys, Ronald L A W; van Diest, Paul J; Moonen, Chrit T W; Mali, Willem P Th M; van den Bosch, Maurice A A J; Bartels, Lambertus W

    2013-10-05

    Magnetic Resonance Imaging-guided High-Intensity Focused Ultrasound (MR-HIFU) is a promising technique for non-invasive breast tumor ablation. The purpose of this study was to investigate the effects of HIFU ablation and thermal exposure on ex vivo human breast tissue. HIFU ablations were performed in three unembalmed cadaveric breast specimens using a clinical MR-HIFU system. Sonications were performed in fibroglandular and adipose tissue. During HIFU ablation, time-resolved anatomical MR images were acquired to monitor macroscopic tissue changes. Furthermore, the breast tissue temperature was measured using a thermocouple to investigate heating and cooling under HIFU exposure. After HIFU ablation, breast tissue samples were excised and prepared for histopathological analysis. In addition, thermal exposure experiments were performed to distinguish between different levels of thermal damage using immunohistochemical staining. Irreversible macroscopic deformations up to 3.7 mm were observed upon HIFU ablation both in fibroglandular and in adipose tissue. No relationship was found between the sonication power or the maximum tissue temperature and the size of the deformations. Temperature measurements after HIFU ablation showed a slow decline in breast tissue temperature. Histopathological analysis of sonicated regions demonstrated ablated tissue and morphologically complete cell death. After thermal exposure, samples exposed to three different temperatures could readily be distinguished. In conclusion, the irreversible macroscopic tissue deformations in ex vivo human breast tissue observed during HIFU ablation suggest that it might be relevant to monitor tissue deformations during MR-HIFU treatments. Furthermore, the slow decrease in breast tissue temperature after HIFU ablation increases the risk of heat accumulation between successive sonications. Since cell death was inflicted after already 5 minutes at 75°C, MR-HIFU may find a place in non-invasive treatment of

  7. Ultrasound-guided tissue fractionation by high intensity focused ultrasound in an in vivo porcine liver model.

    Science.gov (United States)

    Khokhlova, Tatiana D; Wang, Yak-Nam; Simon, Julianna C; Cunitz, Bryan W; Starr, Frank; Paun, Marla; Crum, Lawrence A; Bailey, Michael R; Khokhlova, Vera A

    2014-06-03

    The clinical use of high intensity focused ultrasound (HIFU) therapy for noninvasive tissue ablation has been recently gaining momentum. In HIFU, ultrasound energy from an extracorporeal source is focused within the body to ablate tissue at the focus while leaving the surrounding organs and tissues unaffected. Most HIFU therapies are designed to use heating effects resulting from the absorption of ultrasound by tissue to create a thermally coagulated treatment volume. Although this approach is often successful, it has its limitations, such as the heat sink effect caused by the presence of a large blood vessel near the treatment area or heating of the ribs in the transcostal applications. HIFU-induced bubbles provide an alternative means to destroy the target tissue by mechanical disruption or, at its extreme, local fractionation of tissue within the focal region. Here, we demonstrate the feasibility of a recently developed approach to HIFU-induced ultrasound-guided tissue fractionation in an in vivo pig model. In this approach, termed boiling histotripsy, a millimeter-sized boiling bubble is generated by ultrasound and further interacts with the ultrasound field to fractionate porcine liver tissue into subcellular debris without inducing further thermal effects. Tissue selectivity, demonstrated by boiling histotripsy, allows for the treatment of tissue immediately adjacent to major blood vessels and other connective tissue structures. Furthermore, boiling histotripsy would benefit the clinical applications, in which it is important to accelerate resorption or passage of the ablated tissue volume, diminish pressure on the surrounding organs that causes discomfort, or insert openings between tissues.

  8. In-house Manual Construction of High-Density and High-Quality Tissue Microarrays by Using Homemade Recipient Agarose-Paraffin Blocks.

    Science.gov (United States)

    Kim, Kyu Ho; Choi, Suk Jin; Choi, Yeon Il; Kim, Lucia; Park, In Suh; Han, Jee Young; Kim, Joon Mee; Chu, Young Chae

    2013-06-01

    Self-made tissue punches can be effectively used to punch holes in blank recipient paraffin blocks and extract tissue cores from the donor paraffin blocks for the low-cost construction of tissue microarrays (TMAs). However, variable degrees of section distortion and loss of the tissue cores can occurs during cutting of the TMAs, posing technical problems for in-house manual construction of high-density TMAs. We aimed to update the method for in-house manual TMA construction to improve the quality of high-density TMAs. Blocks of agarose gel were subjected to the standard tissue processing and embedding procedure to prepare recipient agarose-paraffin blocks. The self-made tissue punches and recipient agarose-paraffin blocks were used to construct TMAs, which were completely melted and re-embedded in paraffin to make finished TMA blocks. The donor tissue cores were completely integrated into the surrounding paraffin of the recipient blocks. This method enabled us to construct high-density TMAs with significantly less section distortion or loss of tissue cores during microtomy. Simple and inexpensive construction of high-density and high-quality TMAs can be warranted by using paraffinized agarose gels as recipient blocks.

  9. Plant tissues in 3D via X-ray tomography: simple contrasting methods allow high resolution imaging.

    Directory of Open Access Journals (Sweden)

    Yannick M Staedler

    Full Text Available Computed tomography remains strongly underused in plant sciences despite its high potential in delivering detailed 3D phenotypical information because of the low X-ray absorption of most plant tissues. Existing protocols to study soft tissues display poor performance, especially when compared to those used on animals. More efficient protocols to study plant material are therefore needed. Flowers of Arabidopsis thaliana and Marcgravia caudata were immersed in a selection of contrasting agents used to treat samples for transmission electron microscopy. Grayscale values for floral tissues and background were measured as a function of time. Contrast was quantified via a contrast index. The thick buds of Marcgravia were scanned to determine which contrasting agents best penetrate thick tissues. The highest contrast increase with cytoplasm-rich tissues was obtained with phosphotungstate, whereas osmium tetroxide and bismuth tatrate displayed the highest contrast increase with vacuolated tissues. Phosphotungstate also displayed the best sample penetration. Furthermore, infiltration with phosphotungstate allowed imaging of all plants parts at a high resolution of 3 µm, which approaches the maximum resolution of our equipment: 1.5 µm. The high affinity of phosphotungstate for vasculature, cytoplasm-rich tissue, and pollen causes these tissues to absorb more X-rays than the surrounding tissues, which, in turn, makes these tissues appear brighter on the scan data. Tissues with different brightness can then be virtually dissected from each other by selecting the bracket of grayscale to be visualized. Promising directions for the future include in silico phenotyping and developmental studies of plant inner parts (e.g., ovules, vasculature, pollen, and cell nuclei via virtual dissection as well as correlations of quantitative phenotypes with omics datasets. Therefore, this work represents a crucial improvement of previous methods, allowing new directions of

  10. Plant tissues in 3D via X-ray tomography: simple contrasting methods allow high resolution imaging.

    Science.gov (United States)

    Staedler, Yannick M; Masson, David; Schönenberger, Jürg

    2013-01-01

    Computed tomography remains strongly underused in plant sciences despite its high potential in delivering detailed 3D phenotypical information because of the low X-ray absorption of most plant tissues. Existing protocols to study soft tissues display poor performance, especially when compared to those used on animals. More efficient protocols to study plant material are therefore needed. Flowers of Arabidopsis thaliana and Marcgravia caudata were immersed in a selection of contrasting agents used to treat samples for transmission electron microscopy. Grayscale values for floral tissues and background were measured as a function of time. Contrast was quantified via a contrast index. The thick buds of Marcgravia were scanned to determine which contrasting agents best penetrate thick tissues. The highest contrast increase with cytoplasm-rich tissues was obtained with phosphotungstate, whereas osmium tetroxide and bismuth tatrate displayed the highest contrast increase with vacuolated tissues. Phosphotungstate also displayed the best sample penetration. Furthermore, infiltration with phosphotungstate allowed imaging of all plants parts at a high resolution of 3 µm, which approaches the maximum resolution of our equipment: 1.5 µm. The high affinity of phosphotungstate for vasculature, cytoplasm-rich tissue, and pollen causes these tissues to absorb more X-rays than the surrounding tissues, which, in turn, makes these tissues appear brighter on the scan data. Tissues with different brightness can then be virtually dissected from each other by selecting the bracket of grayscale to be visualized. Promising directions for the future include in silico phenotyping and developmental studies of plant inner parts (e.g., ovules, vasculature, pollen, and cell nuclei) via virtual dissection as well as correlations of quantitative phenotypes with omics datasets. Therefore, this work represents a crucial improvement of previous methods, allowing new directions of research to be

  11. High levels of xanthine oxidoreductase in rat endothelial, epithelial and connective tissue cells. A relation between localization and function?

    NARCIS (Netherlands)

    Kooij, A.; Bosch, K. S.; Frederiks, W. M.; van Noorden, C. J.

    1992-01-01

    The localization of xanthine oxidoreductase activity was investigated in unfixed cryostat sections of various rat tissues by an enzyme histochemical method which specifically demonstrates both the dehydrogenase and oxidase forms of xanthine oxidoreductase. High activity was found in epithelial cells

  12. Influence of High-Fat Diet on Bone Tissue: An Experimental Study in Growing Rats.

    Science.gov (United States)

    Rezende Yanagihara, G; Carminati Shimano, R; Atsuko Tida, J; Suzuki Yamanaka, J; Yasuyo Fukada, S; Mardegan Issa, J P; Shimano, A C; Tavares, J M

    2017-01-01

    The relationship between obesity and bone tissue remains contradictory, especially when the effect of high-fat diet is assessed in experimental models. The aim of this study was to evaluate the effects of high-fat diet on bone metabolism of growing rats. Twenty weaned female Wistar rats were equally divided into two groups: SD (standard diet) and HFD (high-fat diet with 60 % of energy as fat). After five weeks of the two diets, the rats were euthanized, and the liver, blood and bones extracted. The liver was analysed for malondialdehyde (MDA) and reduced glutathione (GSH) concentrations. Blood was analysed by the ELISA method for osteoprotegerin (OPG) and tumour necrosis factor ligand superfamily member 11 (TNFSF11/RANKL). The bone tissue was analysed by dual-energy X-ray absorptiometry (DXA), mechanical tests, computed microtomography, histological quantitative analysis and scanning electron microscopy. The gene expressions of PPAR-γ Runx-2, RANKL and Cathepsin-K were also evaluated. HFD caused an increase in the MDA concentration, indicating oxidative stress. It also increased the expression of PPAR-γ, which is the gene that is related to adipocyte differentiation. There was an increase in BMD of the tibia of animals fed with the HFD, but other microstructural and mechanical properties were maintained unaltered. In addition, there were no changes in the gene expressions related to the differentiation of osteoblasts and osteoclasts, as well as no changes to the biochemical markers of bone formation and bone resorption. Liver and gene parameters are changed in response to the HFD. However, although there was an increase in BMD, the microstructure and function of the bone did not change after a 5-week HFD.

  13. High sensitivity non-invasive detection of calcifications deep inside biological tissue using Transmission Raman Spectroscopy.

    Science.gov (United States)

    Ghita, Adrian; Matousek, Pavel; Stone, Nick

    2018-01-01

    The aim of this research was to develop a novel approach to probe non-invasively the composition of inorganic chemicals buried deep in large volume biological samples. The method is based on advanced Transmission Raman Spectroscopy (TRS) permitting chemical specific detection within a large sampling volume. The approach could be beneficial to chemical identification of the breast calcifications detected during mammographic X-ray procedures. The chemical composition of a breast calcification reflects the pathology of the surrounding tissue, malignant or benign and potentially the grade of malignancy. However, this information is not available from mammography, leading to excisional biopsy and histopathological assessment for a definitive diagnosis. Here we present, for the first time, a design of a new high performance deep Raman instrument and demonstrate its capability to detect type II calcifications (calcium hydroxyapatite) at clinically relevant concentrations and depths of around 40 mm in phantom tissue. This is around double the penetration depth achieved with our previous instrument design and around two orders of magnitude higher than that possible when using conventional Raman spectroscopy. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. High-density polytetrafluoroethylene membranes in guided bone and tissue regeneration procedures: a literature review.

    Science.gov (United States)

    Carbonell, J M; Martín, I Sanz; Santos, A; Pujol, A; Sanz-Moliner, J D; Nart, J

    2014-01-01

    Expanded polytetrafluoroethylene (e-PTFE) has been used successfully as a membrane barrier for regeneration procedures. However, when exposed to the oral cavity, its high porosity increases the risk of early infection, which can affect surgical outcomes. An alternative to e-PTFE is non-expanded and dense polytetrafluoroethylene (n-PFTE), which results in lower levels of early infection following surgical procedures. The aim of this literature review was to analyze and describe the available literature on n-PFTE, report the indications for use, advantages, disadvantages, surgical protocols, and complications. The medical databases Medline-PubMed and Cochrane Library were searched and supplemented with a hand search for reports published between 1980 and May 2012 on n-PTFE membranes. The search strategy was limited to animal, human, and in vitro studies in dental journals published in English. Twenty-four articles that analyzed the use of n-PTFE as a barrier membrane for guided tissue regeneration and guided bone regeneration around teeth and implants were identified: two in vitro studies, seven experimental studies, and 15 clinical studies. There is limited clinical and histological evidence for the use of n-PTFE membranes at present, with some indications in guided tissue regeneration and guided bone regeneration in immediate implants and fresh extraction sockets. Copyright © 2013 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  15. Assessment of subcutaneous adipose tissue using ultrasound in highly trained junior rowers.

    Science.gov (United States)

    Kelso, Anne; Trájer, Emese; Machus, Katja; Treff, Gunnar; Müller, Wolfram; Steinacker, Jürgen M

    2017-06-01

    There is a large variety of body fat (BF) measurements, which differ in validity and reliability. The aim of this study was to measure subcutaneous adipose tissue (SAT) using ultrasound (US) in highly trained junior rowers in a field setting, establish fat patterning profiles, and compare the profiles between male and female athletes. Skinfold thickness (SKF) measurements were also taken and compared to US measurements. Sixteen athletes participated. US measurements were taken at eight sites and reported as a sum of SAT (D): DExcl (without embedded structures) and DIncl (including embedded structures). SKF was measured at three sites and reported as a sum of adipose tissue thickness (SUMSKF). Mean SAT thickness (DIncl) was 27.6 ± 12.4 mm for males and 65.5 ± 11.8 mm for females. Females had significantly more embedded structures than males (P = .016). Significant correlations were found (P < .001, r = 0.92) comparing SUMSKF to DIncl and between SKF and US measurements at the thigh site (P < .001, r = 0.86). US is a suitable tool to measure BF in the field testing of athletes and enables measurements of SAT with an accuracy and reliability not reached before. The sum of thicknesses (DIncl or DExcl) can be used to represent subcutaneous fat based on accurate measurements of uncompressed SAT thicknesses.

  16. High-throughput tissue extraction protocol for NMR- and MS-based metabolomics.

    Science.gov (United States)

    Wu, Huifeng; Southam, Andrew D; Hines, Adam; Viant, Mark R

    2008-01-15

    In metabolomics, tissues typically are extracted by grinding in liquid nitrogen followed by the stepwise addition of solvents. This is time-consuming and difficult to automate, and the multiple steps can introduce variability. Here we optimize tissue extraction methods compatible with high-throughput, reproducible nuclear magnetic resonance (NMR) spectroscopy- and mass spectrometry (MS)-based metabolomics. Previously, we concluded that methanol/chloroform/water extraction is preferable for metabolomics, and we further optimized this here using fish liver and an automated Precellys 24 bead-based homogenizer, allowing rapid extraction of multiple samples without carryover. We compared three solvent addition strategies: stepwise, two-step, and all solvents simultaneously. Then we evaluated strategies for improved partitioning of metabolites between solvent phases, including the addition of extra water and different partition times. Polar extracts were analyzed by NMR and principal components analysis, and the two-step approach was preferable based on lipid partitioning, reproducibility, yield, and throughput. Longer partitioning or extra water increased yield and decreased lipids in the polar phase but caused metabolic decay in these extracts. Overall, we conclude that the two-step method with extra water provides good quality data but that the two-step method with 10 min partitioning provides a more accurate snapshot of the metabolome. Finally, when validating the two-step strategy using NMR and MS metabolomics, we showed that technical variability was considerably smaller than biological variability.

  17. Towards High-Resolution Tissue Imaging Using Nanospray Desorption Electrospray Ionization Mass Spectrometry Coupled to Shear Force Microscopy

    Science.gov (United States)

    Nguyen, Son N.; Sontag, Ryan L.; Carson, James P.; Corley, Richard A.; Ansong, Charles; Laskin, Julia

    2017-07-01

    Constant mode ambient mass spectrometry imaging (MSI) of tissue sections with high lateral resolution of better than 10 μm was performed by combining shear force microscopy with nanospray desorption electrospray ionization (nano-DESI). Shear force microscopy enabled precise control of the distance between the sample and nano-DESI probe during MSI experiments and provided information on sample topography. Proof-of-concept experiments were performed using lung and brain tissue sections representing spongy and dense tissues, respectively. Topography images obtained using shear force microscopy were comparable to the results obtained using contact profilometry over the same region of the tissue section. Variations in tissue height were found to be dependent on the tissue type and were in the range of 0-5 μm for lung tissue and 0-3 μm for brain tissue sections. Ion images of phospholipids obtained in this study are in good agreement with literature data. Normalization of nano-DESI MSI images to the signal of the internal standard added to the extraction solvent allowed us to construct high-resolution ion images free of matrix effects. [Figure not available: see fulltext.

  18. Effects of high-carbohydrate diets on lipogenesis in rat interscapular brown adipose tissue

    OpenAIRE

    Weaire, P. John; Kanagasabai, Tazeen F.

    1982-01-01

    Cycloplasmic preparations from brown and white adipose tissues were assayed for three lipogenic enzymes throughout a programme of starvation followed by refeeding on either a normal or a white-bread diet. In the brown adipose tissue of rats fed on a white-bread diet the three enzymes were elevated to levels significantly higher than those in white adipose tissue.

  19. RNA-Sequencing of Drosophila melanogaster Head Tissue on High-Sugar and High-Fat Diets

    Science.gov (United States)

    Hemphill, Wayne; Rivera, Osvaldo; Talbert, Matthew

    2017-01-01

    Obesity has been shown to increase risk for cardiovascular disease and type-2 diabetes. In addition, it has been implicated in aggravation of neurological conditions such as Alzheimer’s. In the model organism Drosophila melanogaster, a physiological state mimicking diet-induced obesity can be induced by subjecting fruit flies to a solid medium disproportionately higher in sugar than protein, or that has been supplemented with a rich source of saturated fat. These flies can exhibit increased circulating glucose levels, increased triglyceride content, insulin-like peptide resistance, and behavior indicative of neurological decline. We subjected flies to variants of the high-sugar diet, high-fat diet, or normal (control) diet, followed by a total RNA extraction from fly heads of each diet group for the purpose of Poly-A selected RNA-Sequencing. Our objective was to identify the effects of obesogenic diets on transcriptome patterns, how they differed between obesogenic diets, and identify genes that may relate to pathogenesis accompanying an obesity-like state. Gene ontology analysis indicated an overrepresentation of affected genes associated with immunity, metabolism, and hemocyanin in the high-fat diet group, and CHK, cell cycle activity, and DNA binding and transcription in the high-sugar diet group. Our results also indicate differences in the effects of the high-fat diet and high-sugar diet on expression profiles in head tissue of flies, despite the reportedly similar phenotypic impacts of the diets. The impacted genes, and how they may relate to pathogenesis in the Drosophila obesity-like state, warrant further experimental investigation. PMID:29141990

  20. RNA-Sequencing of Drosophila melanogaster Head Tissue on High-Sugar and High-Fat Diets

    Directory of Open Access Journals (Sweden)

    Wayne Hemphill

    2018-01-01

    Full Text Available Obesity has been shown to increase risk for cardiovascular disease and type-2 diabetes. In addition, it has been implicated in aggravation of neurological conditions such as Alzheimer’s. In the model organism Drosophila melanogaster, a physiological state mimicking diet-induced obesity can be induced by subjecting fruit flies to a solid medium disproportionately higher in sugar than protein, or that has been supplemented with a rich source of saturated fat. These flies can exhibit increased circulating glucose levels, increased triglyceride content, insulin-like peptide resistance, and behavior indicative of neurological decline. We subjected flies to variants of the high-sugar diet, high-fat diet, or normal (control diet, followed by a total RNA extraction from fly heads of each diet group for the purpose of Poly-A selected RNA-Sequencing. Our objective was to identify the effects of obesogenic diets on transcriptome patterns, how they differed between obesogenic diets, and identify genes that may relate to pathogenesis accompanying an obesity-like state. Gene ontology analysis indicated an overrepresentation of affected genes associated with immunity, metabolism, and hemocyanin in the high-fat diet group, and CHK, cell cycle activity, and DNA binding and transcription in the high-sugar diet group. Our results also indicate differences in the effects of the high-fat diet and high-sugar diet on expression profiles in head tissue of flies, despite the reportedly similar phenotypic impacts of the diets. The impacted genes, and how they may relate to pathogenesis in the Drosophila obesity-like state, warrant further experimental investigation.

  1. Responsiveness to thyroid hormone and to ambient temperature underlies differences between brown adipose tissue and skeletal muscle thermogenesis in a mouse model of diet-induced obesity.

    Science.gov (United States)

    Ueta, Cintia B; Olivares, Emerson L; Bianco, Antonio C

    2011-09-01

    Thyroid hormone accelerates energy expenditure (EE) and is critical for cold-induced thermogenesis. To define the metabolic role played by thyroid hormone in the dissipation of calories from diet, hypothyroid mice were studied for 60 d in a comprehensive lab animal monitoring system. Hypothyroidism decreased caloric intake and body fat while down-regulating genes in the skeletal muscle but not brown adipose tissue thermogenic programs, without affecting daily EE. Only at thermoneutrality (30 C) did hypothyroid mice exhibit slower rate of EE, indicating a metabolic response to hypothyroidism that depends on ambient temperature. A byproduct of this mechanism is that at room temperature (22 C), hypothyroid mice are protected against diet-induced obesity, i.e. only at thermoneutrality did hypothyroid mice become obese when placed on a high-fat diet (HFD). This is in contrast to euthyroid controls, which on a HFD gained more body weight and fat at any temperature while activating the brown adipose tissue and accelerating daily EE but not the skeletal muscle thermogenic program. In the liver of euthyroid controls, HFD caused an approximately 5-fold increase in triglyceride content and expression of key metabolic genes, whereas acclimatization to 30 C cut triglyceride content by half and normalized gene expression. However, in hypothyroid mice, HFD-induced changes in liver persisted at 30 C, resulting in marked liver steatosis. Acclimatization to thermoneutrality dramatically improves glucose homeostasis, but this was not affected by hypothyroidism. In conclusion, hypothyroid mice are metabolically sensitive to environmental temperature, constituting a mechanism that defines resistance to diet-induced obesity and hepatic lipid metabolism.

  2. Freeze frame analysis on high speed cinematography of Nd/YAG laser explosions in ocular tissues.

    Science.gov (United States)

    Vernon, S A; Cheng, H

    1986-01-01

    High speed colour cinematography at 400 frames per second was used to photograph both single and train burst Nd/YAG laser applications in ox eyes at threshold energy levels. Measurements of the extent and speed of particle scatter and tissue distortion from the acoustic transient were made from a sequential freeze frame analysis of the films. Particles were observed to travel over 8 mm from the site of Nd/YAG application 20 milliseconds after a single pulse at initial speeds in excess of 20 km/h. The use of train bursts of pulses was seen to increase the number of particles scattered and project the wavefront of particles further from the point of laser application. Images PMID:3754458

  3. High-resolution NMR spectroscopy of biological tissues usingprojected Magic Angle Spinning

    Energy Technology Data Exchange (ETDEWEB)

    Martin, Rachel W.; Jachmann, Rebecca C.; Sakellariou, Dimitris; Nielsen, Ulla Gro; Pines, Alexander

    2005-01-27

    High-resolution NMR spectra of materials subject toanisotropic broadening are usually obtained by rotating the sample aboutthe magic angle, which is 54.7 degrees to the static magnetic field. Inprojected Magic Angle Spinning (p-MAS), the sample is spun about twoangles, neither of which is the magic angle. This provides a method ofobtaining isotropic spectra while spinning at shallow angles. The p-MASexperiment may be used in situations where spinning the sample at themagic angle is not possible due to geometric or other constraints,allowing the choice of spinning angle to be determined by factors such asthe shape of the sample, rather than by the spin physics. The applicationof this technique to bovine tissue samples is demonstrated as a proof ofprinciple for future biological or medical applications.

  4. Differential Effects of High-Fish Oil and High-Lard Diets on Cells and Cytokines Involved in the Inflammatory Process in Rat Insulin-Sensitive Tissues

    Directory of Open Access Journals (Sweden)

    Lillà Lionetti

    2014-02-01

    Full Text Available Dietary fat sources may differentially affect the development of inflammation in insulin-sensitive tissues during chronic overfeeding. Considering the anti-inflammatory properties of ω-3 fatty acids, this study aimed to compare the effects of chronic high-fish oil and high-lard diets on obesity-related inflammation by evaluating serum and tissue adipokine levels and histological features in insulin-sensitive tissues (white adipose tissue, skeletal muscle and liver. As expected, a high-lard diet induced systemic and peripheral inflammation and insulin resistance. Conversely, compared with a high-lard diet, a high-fish oil diet resulted in a lower degree of systemic inflammation and insulin resistance that were associated with a lower adipocyte diameter as well as lower immunoreactivity for transforming growth factor β 1 (TGFβ1 in white adipose tissue. A high-fish oil diet also resulted in a lower ectopic lipid depot, inflammation degree and insulin resistance in the skeletal muscle and liver. Moreover, a high-fish oil diet attenuated hepatic stellate cell activation and fibrogenesis in the liver, as indicated by the smooth muscle α-actin (α-SMA and TGFβ1 levels. The replacement of lard (saturated fatty acids with fish oil (ω-3 fatty acids in chronic high-fat feeding attenuated the development of systemic and tissue inflammation.

  5. A TMA de-arraying method for high throughput biomarker discovery in tissue research.

    Directory of Open Access Journals (Sweden)

    Yinhai Wang

    Full Text Available BACKGROUND: Tissue MicroArrays (TMAs represent a potential high-throughput platform for the analysis and discovery of tissue biomarkers. As TMA slides are produced manually and subject to processing and sectioning artefacts, the layout of TMA cores on the final slide and subsequent digital scan (TMA digital slide is often disturbed making it difficult to associate cores with their original position in the planned TMA map. Additionally, the individual cores can be greatly altered and contain numerous irregularities such as missing cores, grid rotation and stretching. These factors demand the development of a robust method for de-arraying TMAs which identifies each TMA core, and assigns them to their appropriate coordinates on the constructed TMA slide. METHODOLOGY: This study presents a robust TMA de-arraying method consisting of three functional phases: TMA core segmentation, gridding and mapping. The segmentation of TMA cores uses a set of morphological operations to identify each TMA core. Gridding then utilises a Delaunay Triangulation based method to find the row and column indices of each TMA core. Finally, mapping correlates each TMA core from a high resolution TMA whole slide image with its name within a TMAMap. CONCLUSION: This study describes a genuine robust TMA de-arraying algorithm for the rapid identification of TMA cores from digital slides. The result of this de-arraying algorithm allows the easy partition of each TMA core for further processing. Based on a test group of 19 TMA slides (3129 cores, 99.84% of cores were segmented successfully, 99.81% of cores were gridded correctly and 99.96% of cores were mapped with their correct names via TMAMaps. The gridding of TMA cores were also extensively tested using a set of 113 pseudo slide (13,536 cores with a variety of irregular grid layouts including missing cores, rotation and stretching. 100% of the cores were gridded correctly.

  6. A TMA de-arraying method for high throughput biomarker discovery in tissue research.

    Science.gov (United States)

    Wang, Yinhai; Savage, Kienan; Grills, Claire; McCavigan, Andrena; James, Jacqueline A; Fennell, Dean A; Hamilton, Peter W

    2011-01-01

    Tissue MicroArrays (TMAs) represent a potential high-throughput platform for the analysis and discovery of tissue biomarkers. As TMA slides are produced manually and subject to processing and sectioning artefacts, the layout of TMA cores on the final slide and subsequent digital scan (TMA digital slide) is often disturbed making it difficult to associate cores with their original position in the planned TMA map. Additionally, the individual cores can be greatly altered and contain numerous irregularities such as missing cores, grid rotation and stretching. These factors demand the development of a robust method for de-arraying TMAs which identifies each TMA core, and assigns them to their appropriate coordinates on the constructed TMA slide. This study presents a robust TMA de-arraying method consisting of three functional phases: TMA core segmentation, gridding and mapping. The segmentation of TMA cores uses a set of morphological operations to identify each TMA core. Gridding then utilises a Delaunay Triangulation based method to find the row and column indices of each TMA core. Finally, mapping correlates each TMA core from a high resolution TMA whole slide image with its name within a TMAMap. This study describes a genuine robust TMA de-arraying algorithm for the rapid identification of TMA cores from digital slides. The result of this de-arraying algorithm allows the easy partition of each TMA core for further processing. Based on a test group of 19 TMA slides (3129 cores), 99.84% of cores were segmented successfully, 99.81% of cores were gridded correctly and 99.96% of cores were mapped with their correct names via TMAMaps. The gridding of TMA cores were also extensively tested using a set of 113 pseudo slide (13,536 cores) with a variety of irregular grid layouts including missing cores, rotation and stretching. 100% of the cores were gridded correctly.

  7. Partial shrinkage of venous tissues near valves using High Intensity Focused Ultrasound

    Science.gov (United States)

    Pichardo, Samuel; Curiel, Laura; Milleret, René; Pichot, Olivier; Lacoste, François; Chapelon, Jean-Yves

    2005-03-01

    The cross-section of a vein can be reduced by exposing the collagen of the vein wall to high temperature (85° C) for a few seconds. Partial shrinkage of the vein is appropriate for correcting deformations of valvular tissues that can cause the abnormal blood reflux which is the main cause of varicose veins and Superficial Venous Insufficiency. Due to its suitability for inducing localized heating, High Intensity Focused Ultrasound (HIFU) is a good method for correcting valvular tissue. In the present study, the feasibility of using HIFU for inducing partial shrinkage of the saphenous vein wall is demonstrated. The position and size of valvular deformations are well suited to being heated and, consequently, reduced with HIFU. The resulting shrinkage of deformations should restore normal function of the valve. An experimental protocol was used in which several in vitro segments of human saphenous vein were exposed with a monochromatic signal produced by a real-time imaging HIFU probe. The probe has a focal length of 45 mm, a diameter of 52.5 mm and operates at 3 MHz. Ultrasonic imaging, obtained with an 8-MHz 128-element linear array placed at the centre of the HIFU probe, was used to target the vein. The segment was inserted in a porcine muscle sample, and both were placed into a PVC cylinder. Individual sonications of the vein wall were performed for acoustic power values ranging between 8.75 and 35 W at a constant sonication duration of 5 s. Different durations ranging between 3 and 7 s at constant power were also tested. Finally, a long duration of 18 s was tested while the focal point was displaced along the vein wall at a speed of 0.5 mm/s. Results showed that shrinkage of the vein wall was observed using echographic and macroscopic analysis. In particular, the vein diameter was reduced by 15% for a sonication-duration of 18 s with continuous displacement of the focal point. Results showed that HIFU is suitable for partial shrinkage of the saphenous vein and

  8. Photoacoustic detection and optical spectroscopy of high-intensity focused ultrasound-induced thermal lesions in biologic tissue

    Energy Technology Data Exchange (ETDEWEB)

    Alhamami, Mosa; Kolios, Michael C.; Tavakkoli, Jahan, E-mail: jtavakkoli@ryerson.ca [Department of Physics, Ryerson University, 350 Victoria Street, Toronto, Ontario M5B 2K3 (Canada)

    2014-05-15

    Purpose: The aims of this study are: (a) to investigate the capability of photoacoustic (PA) method in detecting high-intensity focused ultrasound (HIFU) treatments in muscle tissuesin vitro; and (b) to determine the optical properties of HIFU-treated and native tissues in order to assist in the interpretation of the observed contrast in PA detection of HIFU treatments. Methods: A single-element, spherically concaved HIFU transducer with a centre frequency of 1 MHz was utilized to create thermal lesions in chicken breast tissuesin vitro. To investigate the detectability of HIFU treatments photoacoustically, PA detection was performed at 720 and 845 nm on seven HIFU-treated tissue samples. Within each tissue sample, PA signals were acquired from 22 locations equally divided between two regions of interest within two volumes in tissue – a HIFU-treated volume and an untreated volume. Optical spectroscopy was then carried out on 10 HIFU-treated chicken breast specimens in the wavelength range of 500–900 nm, in 1-nm increments, using a spectrophotometer with an integrating sphere attachment. The authors’ optical spectroscopy raw data (total transmittance and diffuse reflectance) were used to obtain the optical absorption and reduced scattering coefficients of HIFU-induced thermal lesions and native tissues by employing the inverse adding-doubling method. The aforementioned interaction coefficients were subsequently used to calculate the effective attenuation coefficient and light penetration depth of HIFU-treated and native tissues in the wavelength range of 500–900 nm. Results: HIFU-treated tissues produced greater PA signals than native tissues at 720 and 845 nm. At 720 nm, the averaged ratio of the peak-to-peak PA signal amplitude of HIFU-treated tissue to that of native tissue was 3.68 ± 0.25 (mean ± standard error of the mean). At 845 nm, the averaged ratio of the peak-to-peak PA signal amplitude of HIFU-treated tissue to that of native tissue was 3.75

  9. The energy budget, thermogenic capacity and behavior in Swiss mice exposed to a consecutive decrease in temperatures.

    Science.gov (United States)

    Zhao, Zhi-Jun; Chi, Qing-Sheng; Cao, Jing; Han, Ying-Dong

    2010-12-01

    The limitation on sustainable energy intake (SusEI) is important because it establishes the upper energetic limit on the ability of animals to disperse, survive and reproduce. However, there are still arguments about what factors impose that limitation. Thermoregulation in cold environments imposes great energy demands on small mammals. A cold-exposed animal has been suggested to be a model suitable for testing these factors. Here, we examined the changes in food intake and digestible energy intake (DEI) as measures of SusEI, thermogenic capacity and behavioral patterns in Swiss mice exposed to consecutively lower ambient temperatures from 23 to -15°C. Cold-exposed mice showed significant decreases in body mass, fat content of the carcass and body temperature, and increases in DEI compared with controls. The time spent on feeding significantly increased with decreasing temperatures, and time spent on general activity decreased following cold exposure. Resting metabolic rate, nonshivering thermogenesis and serum tri-iodothyronine levels significantly increased in mice exposed to lower temperatures in comparison with controls, whereas these thermogenic variables were not significantly different between 0 and -15°C. It might suggest that SusEI in cold exposed Swiss mice was constrained peripherally by the capacity to produce heat and also by the ability to dissipate body heat, but to a different extent. Moderate cold exposure might result in a relaxation of the heat dissipation limit (HDL), allowing the animals to increase food intake to meet cold stress. When animals are exposed to severe cold, the thermogenenic capacity might reach a ceiling, failing to compensate for the heat loss and which would finally result in lower body temperature and considerable weight loss. This might indicate that the HDL was set at a higher level than peripheral limits for Swiss mice exposed to a consecutive decrease in ambient temperatures.

  10. Affinity imaging mass spectrometry (AIMS): high-throughput screening for specific small molecule interactions with frozen tissue sections.

    Science.gov (United States)

    Yoshimi, T; Kawabata, S; Taira, S; Okuno, A; Mikawa, R; Murayama, S; Tanaka, K; Takikawa, O

    2015-11-07

    A novel screening system, using affinity imaging mass spectrometry (AIMS), has been developed to identify protein aggregates or organ structures in unfixed human tissue. Frozen tissue sections are positioned on small (millimetre-scale) stainless steel chips and incubated with an extensive library of small molecules. Candidate molecules showing specific affinity for the tissue section are identified by imaging mass spectrometry (IMS). As an example application, we screened over a thousand compounds against Alzheimer's disease (AD) brain tissue and identified several compounds with high affinity for AD brain sections containing tau deposits compared to age-matched controls. It should also be possible to use AIMS to isolate chemical compounds with affinity for tissue structures or components that have been extensively modified by events such as oxidation, phosphorylation, acetylation, aggregation, racemization or truncation, for example, due to aging. It may also be applicable to biomarker screening programs.

  11. The gene expression profile of non-cultured, highly purified human adipose tissue pericytes: Transcriptomic evidence that pericytes are stem cells in human adipose tissue

    Energy Technology Data Exchange (ETDEWEB)

    Silva Meirelles, Lindolfo da, E-mail: lindolfomeirelles@gmail.com [Center for Cell-Based Therapy (CEPID/FAPESP), Regional Center for Hemotherapy of Ribeirão Preto, University of São Paulo, Rua Tenente Catão Roxo 2501, 14051-140 Ribeirão Preto, SP (Brazil); Laboratory for Stem Cells and Tissue Engineering, PPGBioSaúde, Lutheran University of Brazil, Av. Farroupilha 8001, 92425-900 Canoas, RS (Brazil); Deus Wagatsuma, Virgínia Mara de; Malta, Tathiane Maistro; Bonini Palma, Patrícia Viana [Center for Cell-Based Therapy (CEPID/FAPESP), Regional Center for Hemotherapy of Ribeirão Preto, University of São Paulo, Rua Tenente Catão Roxo 2501, 14051-140 Ribeirão Preto, SP (Brazil); Araújo, Amélia Goes; Panepucci, Rodrigo Alexandre [Laboratory of Large-Scale Functional Biology (LLSFBio), Regional Center for Hemotherapy of Ribeirão Preto, University of São Paulo, Rua Tenente Catão Roxo 2501, 14051-140 Ribeirão Preto, SP (Brazil); and others

    2016-12-10

    Pericytes (PCs) are a subset of perivascular cells that can give rise to mesenchymal stromal cells (MSCs) when culture-expanded, and are postulated to give rise to MSC-like cells during tissue repair in vivo. PCs have been suggested to behave as stem cells (SCs) in situ in animal models, although evidence for this role in humans is lacking. Here, we analyzed the transcriptomes of highly purified, non-cultured adipose tissue (AT)-derived PCs (ATPCs) to detect gene expression changes that occur as they acquire MSC characteristics in vitro, and evaluated the hypothesis that human ATPCs exhibit a gene expression profile compatible with an AT SC phenotype. The results showed ATPCs are non-proliferative and express genes characteristic not only of PCs, but also of AT stem/progenitor cells. Additional analyses defined a gene expression signature for ATPCs, and revealed putative novel ATPC markers. Almost all AT stem/progenitor cell genes differentially expressed by ATPCs were not expressed by ATMSCs or culture-expanded ATPCs. Genes expressed by ATMSCs but not by ATPCs were also identified. These findings strengthen the hypothesis that PCs are SCs in vascularized tissues, highlight gene expression changes they undergo as they assume an MSC phenotype, and provide new insights into PC biology. - Highlights: • Non-cultured adipose tissue-derived human pericytes (ncATPCs) exhibit a distinctive gene expression signature. • ncATPCs express key adipose tissue stem cell genes previously described in vivo in mice. • ncATPCs express message for anti-proliferative and antiangiogenic molecules. • Most ncATPC-specific transcripts are absent in culture-expanded pericytes or ATMSCs • Gene expression changes ncATPCs undergo as they acquire a cultured ATMSC phenotype are pointed out.

  12. Fabrication of highly aligned fibrous scaffolds for tissue regeneration by centrifugal spinning technology

    Energy Technology Data Exchange (ETDEWEB)

    Loordhuswamy, Amalorpava Mary [Department of Textile Technology, Anna University, Chennai 600025 (India); Krishnaswamy, Venkat Raghavan; Korrapati, Purna Sai [Department of Biomaterials, CSIR-Central Leather Research Institute, Chennai 600020 (India); Thinakaran, Senthilram [Department of Textile Technology, Anna University, Chennai 600025 (India); Rengaswami, Giri Dev Venkateshwarapuram, E-mail: vrgiridev@yahoo.com [Department of Textile Technology, Anna University, Chennai 600025 (India)

    2014-09-01

    Centrifugal spinning (C-Spin) is an emerging technology which uses centrifugal force to produce ultrafine fibers. Being a voltage free technique it can overcome the limitations of electrospinning. Owing to the unique characteristic features such as high surface area to volume ratio, porosity, mechanical strength and fiber alignment, centrifugal spun (C-spun) fibrous mat has a wide range of scope in various biomedical applications. Higher degree of fiber alignment can be effortlessly achieved by the C-Spin process. In order to prove the versatility of C-Spin system with respect to fiber alignment, Polycaprolactone (PCL) and gelatin were spun taking them as model polymers. The morphological analysis revealed that highly aligned ultrafine fibers with smooth surface are achieved by C-Spinning. Hydrophilicity, porosity and mechanical property results confirm that the C-spun mat is more suitable for tissue engineering applications. In vitro and in vivo experiments proved that the scaffolds are biocompatible and can be efficiently used as a wound dressing material. - Highlights: • Highly aligned PCL/gelatin fibrous scaffolds were prepared by C-Spinning system. • Degree of fiber alignment was influenced by the proportion of gelatin in the blends. • Direction of cell growth was parallel to the direction of fiber alignment. • C-Spun matrices can efficiently accelerate faster wound healing.

  13. Tissue contaminants and associated transcriptional response in trout liver from high elevation lakes of Washington

    Science.gov (United States)

    Moran, P.W.; Aluru, N.; Black, R.W.; Vijayan, M.M.

    2007-01-01

    The consistent cold temperatures and large amount of precipitation in the Olympic and Cascade ranges of Washington State are thought to enhance atmospheric deposition of contaminants. However, little is known about contaminant levels in organisms residing in these remote high elevation lakes. We measured total mercury and 28 organochlorine compounds in trout collected from 14 remote lakes in the Olympic, Mt. Rainer, and North Cascades National Parks. Mercury was detected in trout from all lakes sampled (15 to 262 ??g/kg ww), while two organochlorines, total polychlorinated biphenyls (tPCB) and dichlorodiphenyldichloroethylene (DDE), were also detected in these fish tissues (response in the liver using a custom-made low-density targeted rainbow trout cDNA microarray. We detected significant differences in liver transcriptional response, including significant changes in metabolic, endocrine, and immune-related genes, in fish collected from Wilcox Lake compared to Skymo Lake. Overall, our results suggest that local urban areas contribute to the observed contaminant patterns in these high elevation lakes, while the transcriptional changes point to a biological response associated with exposure to these contaminants in fish. Specifically, the gene expression pattern leads us to hypothesize a role for mercury in disrupting the metabolic and reproductive pathways in fish from high elevation lakes in western Washington. ?? 2007 American Chemical Society.

  14. Central efferent pathways mediating skin cooling-evoked sympathetic thermogenesis in brown adipose tissue

    OpenAIRE

    Nakamura, Kazuhiro; Morrison, Shaun F.

    2006-01-01

    Control of thermoregulatory effectors by the autonomic nervous system is a critical component of rapid cold-defense responses, which are triggered by thermal information from the skin. However, the central autonomic mechanism driving thermoregulatory effector responses to skin thermal signals remains to be determined. Here, we examined the involvement of several autonomic brain regions in sympathetic thermogenic responses in brown adipose tissue (BAT) to skin cooling in urethane-chloralose-an...

  15. Brown adipose tissue in the treatment of obesity and diabetes: Are we hot enough?

    OpenAIRE

    Tan, Chong Yew; Ishikawa, Ko; Virtue, Samuel; Vidal‐Puig, Antonio

    2011-01-01

    Abstract The identification of functional brown adipose tissue in human adults has intensified interest in exploiting thermogenic energy expenditure for the purpose of weight management. However, food intake and energy expenditure are tightly regulated and it is generally accepted that variation in one component results in compensatory changes in the other. In the context of weight loss, additional biological adaptations occur in an attempt to further limit weight loss. In the present review,...

  16. Effects of Endogenous Oxytocin Receptor Signaling in Nucleus Tractus Solitarius on Satiation-Mediated Feeding and Thermogenic Control in Male Rats.

    Science.gov (United States)

    Ong, Zhi Yi; Bongiorno, Diana M; Hernando, Mary Ann; Grill, Harvey J

    2017-09-01

    Central oxytocin receptor (OT-R) signaling reduces food intake and increases energy expenditure, but the central sites and mechanisms mediating these effects are unresolved. We showed previously that pharmacological activation of OT-R in hindbrain/nucleus tractus solitarius (NTS) amplifies the intake-inhibitory effects of gastrointestinal (GI) satiation signals. Unexplored were the energetic effects of hindbrain OT-R agonism and the physiological relevance of NTS OT-R signaling on food intake and energy expenditure control. Using a virally mediated OT-R knockdown (KD) strategy and a range of behavioral paradigms, this study examined the role of endogenous NTS OT-R signaling on satiation-mediated food intake inhibition and thermogenic control. Results showed that, compared with controls, NTS OT-R KD rats consumed larger meals, were less responsive to the intake-inhibitory effects of a self-ingested preload, and consumed more chow following a 24-hour fast. These data indicate that NTS OT-R signaling is necessary for normal satiation control. Whereas both control and NTS OT-R KD rats increased core temperature following high-fat diet maintenance (relative to chow maintenance), the percent increase in core temperature was greater in control compared with NTS OT-R KD rats during the light cycle. Hindbrain oxytocin agonist delivery increased core temperature in both control and NTS OT-R KD rats and the percent increase relative to vehicle treatment was not significantly different between groups. Together, data reveal a critical role for endogenous NTS OT-R signaling in mediating the intake-inhibitory effects of endogenous GI satiation signals and in diet-induced thermogenesis. Copyright © 2017 Endocrine Society.

  17. Differential expression of aquaporin 7 in adipose tissue of lean and obese high fat consumers.

    Science.gov (United States)

    Marrades, M Pilar; Milagro, Fermin I; Martínez, J Alfredo; Moreno-Aliaga, Maria J

    2006-01-20

    Aquaporin 7 (AQP7) is an aquaglyceroprotein responsible for the secretion and uptake of glycerol from the adipocyte. The modulation of the expression of this membrane transport protein might play an important role in the susceptibility to the development of obesity. The aim of the present study was to compare the AQP7 gene expression in subcutaneous abdominal fat in lean vs. obese high fat intakers with a similar daily physical activity pattern. Twelve young men, 6 lean (BMI=23.2+/-0.4kg/m(2)) and 6 obese (35.0+/-1.1kg/m(2)) with a similar habitual dietary intake of fat (45.5+/-2.5 vs. 43.5+/-1.7% daily energy from fat for lean and obese, respectively) and physical activity (16.0+/-5.7 vs. 17.2+/-5.1 METsh/week for lean and obese, respectively), were recruited. Subcutaneous abdominal fat biopsies were obtained and total RNA was extracted and purified. Pools of RNA from lean and obese individuals were probed into Affymetrix GeneChip Human U133A. The microarray analysis revealed that AQP7 gene was down-regulated in obese compared to lean subjects. The results of the microarray analysis were confirmed by real-time PCR studies. In summary, our data show that the AQP7 gene is differentially expressed in adipose tissue of lean and obese individuals. The down-regulation of the AQP7 gene could be implicated in the susceptibility to obesity by reducing glycerol release and promoting the accumulation of lipids in the adipose tissue.

  18. ISDoT: in situ decellularization of tissues for high-resolution imaging and proteomic analysis of native extracellular matrix

    DEFF Research Database (Denmark)

    Mayorca-Guiliani, Alejandro E.; Madsen, Chris D.; Cox, Thomas R.

    2017-01-01

    decellularization of tissues (ISDoT), it allows whole organs to be decellularized, leaving native ECM architecture intact. These three-dimensional decellularized tissues can be studied using high-resolution fluorescence and second harmonic imaging, and can be used for quantitative proteomic interrogation of the ECM....... We also describe differing patterns of basement-membrane organization surrounding different types of blood vessels in healthy and diseased tissues. The ISDoT procedure allows for the study of native ECM structure under normal and pathological conditions in unprecedented detail....

  19. Glycemic index differences of high-fat diets modulate primarily lipid metabolism in murine adipose tissue.

    Science.gov (United States)

    van Schothorst, Evert M; Bunschoten, Annelies; Verlinde, Eline; Schrauwen, Patrick; Keijer, Jaap

    2011-08-16

    A low vs. high glycemic index of a high-fat (HF) diet (LGI and HGI, respectively) significantly retarded adverse health effects in adult male C57BL/6J mice, as shown recently (Van Schothorst EM, Bunschoten A, Schrauwen P, Mensink RP, Keijer J. FASEB J 23: 1092-1101, 2009). The LGI diet enhanced whole body insulin sensitivity and repressed HF diet-induced body and white adipose tissue (WAT) weight gain, resulting in significantly reduced serum leptin and resistin levels and increased adiponectin levels. We questioned how WAT is modulated and characterized the molecular mechanisms underlying the glycemic index-mediated effects using whole genome microarrays. This showed that the LGI diet mainly exerts its beneficial effects via substrate metabolism, especially fatty acid metabolism. In addition, cell adhesion and cytoskeleton remodeling showed reduced expression, in line with lower WAT mass. An important transcription factor showing enhanced expression is PPAR-γ. Furthermore, serum levels of triglycerides, total cholesterol, and HDL- and LDL-cholesterol were all significantly reduced by LGI diet, and simultaneously muscle insulin sensitivity was significantly increased as analyzed by protein kinase B/Akt phosphorylation. Cumulatively, even though these mice were fed an HF diet, the LGI diet induced significantly favorable changes in metabolism in WAT. These effects suggest a partial overlap with pharmacological approaches by thiazolidinediones to treat insulin resistance and statins for hypercholesterolemia. It is therefore tempting to speculate that such a dietary approach might beneficially support pharmacological treatment of insulin resistance or hypercholesterolemia in humans.

  20. Tissue storage and control of cholesterol metabolism in man on high cholesterol diets.

    Science.gov (United States)

    Quintão, E C; Brumer, S; Stechhahn, K

    1977-03-01

    The possibility of accumulation of tissue cholesterol in human beings submitted to high cholesterol feeding was investigated in liver biopsies and through fecal sterol balance studies. Feeding to 10 individuals 3.1 to 3.4 g/day of cholesterol for 3 weeks raised the mean serum level from 293 to 349 mg/100 ml, namely 19%, whereas the liver cholesterol content was 417 mg/100 g of wet weight. In 10 control cases eating 0.1--0.4 g/day of cholesterol serum cholesterol remained stable throughout the experimental period and the liver cholesterol content was 256 mg/100 g. Difference of liver colesterol level between the two groups was 62%. In 7 patients submitted to two periods of balance investigation on a cholesterol-free synthetic formula diet respectively prior to (PI) and after (PIII) eating the high cholesterol solid food from 4 to 15 weeks (PII), fecal steroid excretion in PIII exceeded PI in 3 patients. Such data are a direct evidence for the existence of an efficient system to release acutely stored cholesterol. In one patient bile acid excretion accounted for the difference between PIII and PI.

  1. Raspberry seed flour attenuates high-sucrose diet-mediated hepatic stress and adipose tissue inflammation.

    Science.gov (United States)

    Kang, Inhae; Espín, Juan Carlos; Carr, Timothy P; Tomás-Barberán, Francisco A; Chung, Soonkyu

    2016-06-01

    Chronic intake of high sucrose (HS) diet exacerbates high-fat (HF) diet-induced obesity and its associated metabolic complications. Previously, we have demonstrated that ellagic acid (EA), an abundant polyphenol found in some fruits and nuts, exerts distinct lipid-lowering characteristics in hepatocytes and adipocytes. In this study, we hypothesized that EA supplementation inhibits HS diet-mediated hepatic toxicity and its accompanied metabolic dysregulation. To test this hypothesis, C57BL/6 male mice were randomly assigned to three isocaloric HF diets (41% calories from fat) containing either no-sucrose (HF), high-sucrose (HFHS), or high-sucrose plus EA (HFHS-R) from raspberry seed flour (RSF, equivalent to 0.03% of EA), and fed for 12weeks. The inclusion of EA from RSF significantly improved HFHS diet-mediated dyslipidemia and restored glucose homeostasis levels similar to the HF diet-fed mice. Despite marginal difference in hepatic triglyceride content, the addition of EA substantially reversed the activation of endoplasmic reticulum (ER) stress and oxidative damage triggered by HFHS diet in the liver. These effects of EA were further confirmed in human hepatoma cells by reducing ER stress and reactive oxygen species (ROS) production. Moreover, HFHS-R diet significantly decreased visceral adipocyte hypertrophy and adipose tissue inflammation evidenced by reduced proinflammatory gene expression and macrophage infiltration. In summary, EA supplementation from RSF was effective in reducing HFHS diet-mediated metabolic complication by attenuating hepatic ER and oxidative stresses as well as adipocyte inflammation. Our results suggest that the inclusion of EA in diets may normalize metabolic insults triggered by HS consumption. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Plant Tissues in 3D via X-Ray Tomography: Simple Contrasting Methods Allow High Resolution Imaging

    OpenAIRE

    Yannick M Staedler; David Masson; Jürg Schönenberger

    2013-01-01

    Computed tomography remains strongly underused in plant sciences despite its high potential in delivering detailed 3D phenotypical information because of the low X-ray absorption of most plant tissues. Existing protocols to study soft tissues display poor performance, especially when compared to those used on animals. More efficient protocols to study plant material are therefore needed. Flowers of Arabidopsis thaliana and Marcgravia caudata were immersed in a selection of contrasting agents ...

  3. Survival of Highly Pathogenic Avian Influenza H5N1 Virus in Tissues Derived from Experimentally Infected Chickens.

    Science.gov (United States)

    Yamamoto, Yu; Nakamura, Kikuyasu; Mase, Masaji

    2017-08-15

    Eurasian lineage highly pathogenic avian influenza (HPAI) H5N1 virus has been a severe threat to the poultry industry since its emergence in 1996. The carcass or tissues derived from infected birds may present the risk of the virus spreading to humans, animals, and the surrounding environment. In this study, we investigated the survival of the virus in feather, muscle, and liver tissues collected from six chickens (Gallus gallus) experimentally infected with HPAI H5N1 virus. The tissues were stored at +4°C or +20°C, and viral isolation was performed at different times for 360 days. The maximum periods for viral survival were observed in samples stored at +4°C in all tissue types and were 240 days in feather tissues, 160 days in muscle, and 20 days in liver. The viral infectivity at +20°C was maintained for a maximum of 30 days in the feather tissues, 20 days in muscle, and 3 days in liver. The viral inactivation rates partly overlapped in the feather and muscle tissues at the two temperatures. The virus was inactivated rapidly in the liver. Our experimental results indicate that the tissue type and temperature can greatly influence the survival of HPAI H5N1 virus in the tissues of infected chickens.IMPORTANCE Highly pathogenic avian influenza virus of the H5N1 subtype can cause massive losses of poultry, and people need to handle a large number of chicken carcasses contaminated with the virus at outbreak sites. This study evaluated how long the virus can keep its infectivity in the three types of tissues derived from chickens infected with the virus. Our experimental results indicate that the virus can survive in tissues for a specific period of time depending on the tissue type and temperature. Our results are valuable for better understanding of viral ecology in the environment and for reducing the risk of the virus spreading via bird tissues contaminated with the virus. Copyright © 2017 American Society for Microbiology.

  4. High-load preconditioning of human soft tissue hamstring grafts: An in vitro biomechanical analysis.

    Science.gov (United States)

    Lockwood, W Charles; Marchetti, Daniel Cole; Dahl, Kimi D; Mikula, Jacob D; Williams, Brady T; Kheir, Matthew M; Turnbull, Travis Lee; LaPrade, Robert F

    2017-01-01

    and the high-load/short-duration protocol, implying that the latter could potentially induce the same viscoelastic changes in soft tissue grafts as the current clinical practice. The "600 N for 20 s" preconditioning protocol may provide similar postoperative results as the clinical protocol, "89 N for 15 min", and also reduce or maintain operative time. A high-load preconditioning protocol that reduces graft elongation may benefit patients undergoing ACL reconstruction, especially for cases of failed primary reconstruction, genu recurvatum, and increased tibial slope, where maintaining graft length is imperative to restore knee stability.

  5. Next Generation Molecular Histology Using Highly Multiplexed Ion Beam Imaging (MIBI) of Breast Cancer Tissue Specimens for Enhanced Clinical Guidance

    Science.gov (United States)

    2016-07-01

    AWARD NUMBER: W81XWH- 14-1-0192 TITLE: Next-Generation Molecular Histology Using Highly Multiplexed Ion Beam Imaging (MIBI) of Breast Cancer...DATES COVERED 4. TITLE AND SUBTITLE Next-Generation Molecular Histology Using Highly Multiplexed Ion Beam Imaging (MIBI) of Breast Cancer Tissue

  6. Adipose tissue biglycan as a potential anti-inflammatory target of sodium salicylate in mice fed a high fat diet

    Directory of Open Access Journals (Sweden)

    Adapala Venkata J

    2012-04-01

    Full Text Available Abstract Background Inflammation in adipose tissue (AT during obesity causes impaired AT function. Although multiple extracellular matrix (ECM proteins are expressed in AT their potential role in adipose tissue inflammation is unclear. Biglycan, a pro-inflammatory ECM gene, is highly enriched in adipose tissue. However, whether it is correlated with adipose tissue inflammation is unknown. We provide evidence in support of a strong association between biglycan expression and inflammatory status of adipose tissue. Methods C57BL6 mice were fed either a control (10% fat calories or a high fat diet (HFD (60% fat calories for 8 weeks. Adipose tissue was analyzed for the expression of biglycan, IL-6 and TNFα. Biglycan knockout or wild type were also fed a high fat diet for 8 weeks and the expression of inflammatory genes in the mesenteric adipose tissue was examined. To test anti-inflammatory treatment on biglycan expression, a group of mice were fed either the low fat or high fat diet for eight weeks supplemented with either saline or sodium salicylate @ 25mg/100ml in their drinking water. Results Mice on HFD had an increase in ECM genes (BGN and COL1A1, inflammatory genes (IL-6 and TNFα in both the subcutaneous and epididymal depots. However, correlation analysis only shows a positive correlation between biglycan, IL-6 and TNFα expression. In addition, lower expression of IL-6 and CD68 was found in the mesenteric adipose tissue of biglycan knockout mice compared to the wild type. Sodium salicylate treatment reduced subcutaneous adipose tissue expression of BGN, COL1A1, and COL6A1 and a concurrent downregulation of TNFα and IL-6 and TLR4 expression. Salicylate also lowered the serum TGFβ1 levels. Conclusion Biglycan expression correlates with adipose tissue inflammation, especially in the subcutaneous depot compared to the epididymal depot. This is supported by the greater effect of sodium salicylate in attenuating both inflammatory and ECM gene

  7. High-throughput immunophenotyping of 43 ferret lymphomas using tissue microarray technology

    DEFF Research Database (Denmark)

    Hammer, Anne Sofie; Williams, B.; Dietz, H.H.

    2007-01-01

    To validate the use of the tissue microarray (TMA) method for immunophenotyping of ferret lymphomas, a TMA was constructed containing duplicate 1-mm cores sampled from 112 paraffin-embedded lymphoma tissue specimens obtained from 43 ferret lymphoma cases. Immunohistochemical (IHC) expression of CD3...

  8. Gender-Specific Mechanisms Underlying the Amelioration of High-Fat Diet-Induced Glucose Intolerance in B-Cell-Activating Factor Deficient Mice.

    Directory of Open Access Journals (Sweden)

    Bobae Kim

    Full Text Available It has recently been found that B cell activating factor (BAFF plays an important role in the regulation of energy homeostasis. We also have previously reported that BAFF deficiency reverses high-fat (HF diet-induced glucose intolerance by potentiating adipose tissue function. In the present study, we found that BAFF deficient (BAFF-/- mice exhibit gender-specific differences in protection against diet-induced glucose intolerance, and aimed to characterize the gender-dependent molecular alterations in energy metabolism. Under HF feeding conditions, serum BAFF level of female wild-type (WT mice was considerably higher than that of male mice. Despite increased body weight gain, both male and female BAFF-/- mice showed significantly improved glucose tolerance compared to their WT counterparts. Expressions of genes involved in glucose transport, thermogenesis and lipid oxidation were up-regulated in brown adipose tissues of both male and female BAFF-/- mice. Interestingly, the expression of thermogenic genes in subcutaneous adipose tissue was significantly enhanced in female BAFF-/- compared to WT mice, but the difference was not observed between male BAFF-/- and WT mice. The enhanced thermogenic program was confirmed by higher protein levels of UCP1 and irisin in female BAFF-/- than in WT mice. Additionally, adiponectin production in white adipose tissues and AMPK phosphorylation in subcutaneous adipose tissue were also significantly elevated in female BAFF-/- compared to WT mice, but not in male BAFF-/- mice. Our findings define a comprehensive scenario for the enhancing effect of BAFF depletion on glucose tolerance wherein the underlying mechanism is, at least in part, gender-specific, and suggest that gender difference should be considered as an important factor in the use of BAFF blockade as a therapeutic approach for the prevention and treatment of type 2 diabetes.

  9. The apoptotic effect of a high dose of toluene on liver tissue during the acute phase: an experimental study.

    Science.gov (United States)

    Ayan, Murat; Tas, Ufuk; Sogut, Erkan; Kuloglu, Tuncay; Cayli, Sevil; Kocaman, Nevin; Karaca, Zafer Ismail; Sahin, Mehmet

    2013-09-01

    The aim of this study is to investigate the acute toxic effects of high-dose toluene and its mechanisms on the liver tissue of toluene-treated rats. In this study, 16 adult male Wistar albino rats (200-220 g) were divided into two equal groups. Group I was used as a control group, while group II was exposed to high dose of toluene, 5200 mg/kg (6 ml/kg per gavage). After the 3-hour experimental period, blood samples and liver tissues were taken from the euthanized animals. Serum aspartate and alanine aminotransferase levels were assayed. Liver tissues were fixed in 10% neutral formalin, then embedded in paraffin and sectioned (5 μm thickness). Sections were stained with hematoxylin and eosin for histopathological examination. A terminal transferase dUTP nick end labeling assay was also done for the determination of apoptosis in liver tissues. For the determination of Bax and caspase-3 immunoreactivity, the sections were stained using avidin-biotin-peroxidase immunohistochemical method. The level of plasma transaminase was found to be increased in toluene administered rats. Additionally, slight degeneration of hepatocyte and mononuclear cell infiltration was observed in the liver tissue sections and a high (+++) immunoreactivity for Bax and caspase-3 protein was observed in the toluene group. This study showed that the high dose of toluene triggers apoptosis in the liver of rats via the mitochondrial pathway in acute period.

  10. Automated breast tissue density assessment using high order regional texture descriptors in mammography

    Science.gov (United States)

    Law, Yan Nei; Lieng, Monica Keiko; Li, Jingmei; Khoo, David Aik-Aun

    2014-03-01

    Breast cancer is the most common cancer and second leading cause of cancer death among women in the US. The relative survival rate is lower among women with a more advanced stage at diagnosis. Early detection through screening is vital. Mammography is the most widely used and only proven screening method for reliably and effectively detecting abnormal breast tissues. In particular, mammographic density is one of the strongest breast cancer risk factors, after age and gender, and can be used to assess the future risk of disease before individuals become symptomatic. A reliable method for automatic density assessment would be beneficial and could assist radiologists in the evaluation of mammograms. To address this problem, we propose a density classification method which uses statistical features from different parts of the breast. Our method is composed of three parts: breast region identification, feature extraction and building ensemble classifiers for density assessment. It explores the potential of the features extracted from second and higher order statistical information for mammographic density classification. We further investigate the registration of bilateral pairs and time-series of mammograms. The experimental results on 322 mammograms demonstrate that (1) a classifier using features from dense regions has higher discriminative power than a classifier using only features from the whole breast region; (2) these high-order features can be effectively combined to boost the classification accuracy; (3) a classifier using these statistical features from dense regions achieves 75% accuracy, which is a significant improvement from 70% accuracy obtained by the existing approaches.

  11. High throughput micro-well generation of hepatocyte micro-aggregates for tissue engineering.

    Science.gov (United States)

    Gevaert, Elien; Dollé, Laurent; Billiet, Thomas; Dubruel, Peter; van Grunsven, Leo; van Apeldoorn, Aart; Cornelissen, Ria

    2014-01-01

    The main challenge in hepatic tissue engineering is the fast dedifferentiation of primary hepatocytes in vitro. One successful approach to maintain hepatocyte phenotype on the longer term is the cultivation of cells as aggregates. This paper demonstrates the use of an agarose micro-well chip for the high throughput generation of hepatocyte aggregates, uniform in size. In our study we observed that aggregation of hepatocytes had a beneficial effect on the expression of certain hepatocyte specific markers. Moreover we observed that the beneficial effect was dependent on the aggregate dimensions, indicating that aggregate parameters should be carefully considered. In a second part of the study, the selected aggregates were immobilized by encapsulation in methacrylamide-modified gelatin. Phenotype evaluations revealed that a stable hepatocyte phenotype could be maintained during 21 days when encapsulated in the hydrogel. In conclusion we have demonstrated the beneficial use of micro-well chips for hepatocyte aggregation and the size-dependent effects on hepatocyte phenotype. We also pointed out that methacrylamide-modified gelatin is suitable for the encapsulation of these aggregates.

  12. High throughput micro-well generation of hepatocyte micro-aggregates for tissue engineering.

    Directory of Open Access Journals (Sweden)

    Elien Gevaert

    Full Text Available The main challenge in hepatic tissue engineering is the fast dedifferentiation of primary hepatocytes in vitro. One successful approach to maintain hepatocyte phenotype on the longer term is the cultivation of cells as aggregates. This paper demonstrates the use of an agarose micro-well chip for the high throughput generation of hepatocyte aggregates, uniform in size. In our study we observed that aggregation of hepatocytes had a beneficial effect on the expression of certain hepatocyte specific markers. Moreover we observed that the beneficial effect was dependent on the aggregate dimensions, indicating that aggregate parameters should be carefully considered. In a second part of the study, the selected aggregates were immobilized by encapsulation in methacrylamide-modified gelatin. Phenotype evaluations revealed that a stable hepatocyte phenotype could be maintained during 21 days when encapsulated in the hydrogel. In conclusion we have demonstrated the beneficial use of micro-well chips for hepatocyte aggregation and the size-dependent effects on hepatocyte phenotype. We also pointed out that methacrylamide-modified gelatin is suitable for the encapsulation of these aggregates.

  13. Simultaneous observation of cavitation bubbles generated in biological tissue by high-speed optical and acoustic imaging methods

    Science.gov (United States)

    Suzuki, Kai; Iwasaki, Ryosuke; Takagi, Ryo; Yoshizawa, Shin; Umemura, Shin-ichiro

    2017-07-01

    Acoustic cavitation bubbles are useful for enhancing the heating effect in high-intensity focused ultrasound (HIFU) treatment. Many studies were conducted to investigate the behavior of such bubbles in tissue-mimicking materials, such as a transparent gel phantom; however, the detailed behavior in tissue was still unclear owing to the difficulty in optical observation. In this study, a new biological phantom was developed to observe cavitation bubbles generated in an optically shallow area of tissue. Two imaging methods, high-speed photography using light scattering and high-speed ultrasonic imaging, were used for detecting the behavior of the bubbles simultaneously. The results agreed well with each other for the area of bubble formation and the temporal change in the region of bubbles, suggesting that both methods are useful for visualizing the bubbles.

  14. Fabrication and design of bioactive agent coated, highly-aligned electrospun matrices for nerve tissue engineering: Preparation, characterization and application

    Science.gov (United States)

    Lee, Sang Jin; Heo, Min; Lee, Donghyun; Heo, Dong Nyoung; Lim, Ho-Nam; Kwon, Il Keun

    2017-12-01

    In this study, we designed highly-aligned thermoplastic polycarbonate urethane (PCU) fibrous scaffolds coated with bioactive compounds, such as Poly-L-Lysine (PLL) and Poly-L-Ornithine (PLO), to enhance cellular adhesion and directivity. These products were characterized by scanning electron microscope (SEM) analysis which demonstrated that highly aligned fiber strands were formed without beads when coated onto a mandrel rotating at 1800 rpm. During in vitro cell test, PLO-coated, aligned PCU scaffolds were found to have significantly higher proliferation rates than PLL coated and bare PCU scaffolds. Interestingly, dental pulp stem cells (DPSCs) were observed to stretch along the longitudinal axis parallel to the cell direction on highly aligned scaffolds. These results clearly confirm that our strategy may suggest a useful paradigm by inducing neural tissue repair as a means to remodeling and healing of tissue for restorative procedures in neural tissue engineering.

  15. The Effects of High Doses of Nandrolone Decanoate on Cardiac Muscle Tissue

    Directory of Open Access Journals (Sweden)

    Sretenovic Jasmina

    2016-12-01

    Full Text Available In recent decades, steroid abuse has become very popular and widespread among professional and recreational athletes. The aim of this study was to examine the chronic effects of training combined with high doses of nandrolone decanoate on cardiac muscle tissue. The study included 32 Wistar albino rats divided into 4 groups: control (T-N-, steroid (T-N+, exercisetraining (T+N- and exercise plus steroid (T+N+ groups. The T+N- and T+N+ group swam for 4 weeks, 1 hour per day, 5 days per week. The N+ (nandrolone positive groups received nandrolone decanoate (20 mg/kg once per week, subcutaneously. After 4 weeks of training, the rats were sacrificed. Heart biopsy specimens were routinely fixed and embedded in paraffin. Fivemicrometre thick sections were stained with haematoxylin and eosin (H/E and Masson-Trichrome dyes. Captured microscopic images were processed by special software for image analysis to quantify results. Our results showed that the combination of nandrolone and training causes left ventricular wall thickening of 30%. Average cardiac muscle cell longitudinal diameter was increased by 6% in the T-N+ group, by 16% in the T+N- group and by 25% in the T+N+ group. The cross sectional muscle cell area was increased in the T+N+ group by 33%. Heart collagen content was increased in the nandrolone group compared to the control group by 261%. Collagen content was decreased in the T+N+ group by 34%. High doses of AAS induced left ventricle hypertrophy and excessive heart collagen deposition.

  16. High dietary sodium intake increases white adipose tissue mass and plasma leptin in rats.

    Science.gov (United States)

    Fonseca-Alaniz, Miriam H; Brito, Luciana C; Borges-Silva, Cristina N; Takada, Julie; Andreotti, Sandra; Lima, Fabio B

    2007-09-01

    Salt restriction has been reported to increase white adipose tissue (WAT) mass in rodents. The objective of this study was to investigate the effect of different sodium content diets on the lipogenic and lipolytic activities of WAT. Male Wistar rats were fed on normal-sodium (NS; 0.5% Na(+)), high-sodium (HS; 3.12% Na(+)), or low-sodium (LS; 0.06% Na(+)) diets for 3, 6, and 9 weeks after weaning. Blood pressure (BP) was measured using a computerized tail-cuff system. At the end of each period, rats were killed and blood samples were collected for leptin determinations. The WAT from abdominal and inguinal subcutaneous (SC), periepididymal (PE) and retroperitoneal (RP) depots was weighed and processed for adipocyte isolation, rate measurement of lipolysis and d-[U-(14)C]-glucose incorporation into lipids, glucose-6-phosphate dehydrogenase (G6PDH) and malic enzyme activity evaluation, and determination of G6PDH and leptin mRNA expression. After 6 weeks, HS diet significantly increased BP; SC, PE, and RP WAT masses; PE adipocyte size; plasma leptin concentration; G6PDH activity in SC WAT; and PE depots and malic activity only in SC WAT. The leptin levels correlated positively with WAT masses and adipocyte size. An increase in the basal and isoproterenol-stimulated lipolysis and in the ability to incorporate glucose into lipids was observed in isolated adipocytes from HS rats. HS diet induced higher adiposity characterized by high plasma leptin concentration and adipocyte hypertrophy, probably due to an increased lipogenic capacity of WAT.

  17. Use of flow cytometry for high-throughput cell population estimates in fixed brain tissue

    Directory of Open Access Journals (Sweden)

    Nicole A Young

    2012-07-01

    Full Text Available The numbers and types of cells in an area of cortex define its function. Therefore it is essential to characterize the numbers and distributions of total cells in areas of the cortex, as well as to identify numbers of subclasses of neurons and glial cells. To date, the large size of the primate brain and the lack of innovation in cell counting methods have been a roadblock to obtaining high-resolution maps of cell and neuron density across the cortex in humans and non-human primates. Stereological counting methods and the isotropic fractionator are valuable tools for estimating cell numbers, but are better suited to smaller, well-defined brain structures or to cortex as a whole. In the present study, we have extended our flow-cytometry based counting method, the flow fractionator (Collins et al., 2010a, to include high-throughput total cell population estimates in homogenized cortical samples. We demonstrate that our method produces consistent, accurate and repeatable cell estimates quickly. The estimates we report are in excellent agreement with estimates for the same samples obtained using a Neubauer chamber and a fluorescence microscope. We show that our flow cytometry-based method for total cell estimation in homogenized brain tissue is more efficient and more precise than manual counting methods. The addition of automated nuclei counting to our flow fractionator method allows for a fully automated, rapid characterization of total cells and neuronal and non-neuronal populations in human and non-human primate brains, providing valuable data to further our understanding of the functional organization of normal, aging and diseased brains.

  18. Disparate effects of feeding on core body and adipose tissue temperatures in animals selectively bred for Nervous or Calm temperament.

    Science.gov (United States)

    Henry, Belinda A; Blache, Dominique; Rao, Alexandra; Clarke, Iain J; Maloney, Shane K

    2010-09-01

    In addition to homeostatic regulation of body mass, nonhomeostatic factors impact on energy balance. Herein we describe effects of temperament on adipose and core body temperatures in sheep. Animals were genetically selected for Nervous or Calm traits. We characterized the effects of 1) high- and low-energy intake and maintenance feeding, 2) meal anticipation, and 3) adrenocorticotropin challenge on core body and adipose temperatures. Temperature measurements (5 min) were made using a thermistor inserted into the carotid artery (core body) and a probe in the retroperitoneal fat. An imposed feeding window was used to establish postprandial elevations in temperature. Fat tissue was taken from retroperitoneal and subcutaneous regions for real-time PCR analyses. We demonstrate that innate differences in temperament impact on adipose and core body temperatures in response to various dietary and evocative stimuli. In response to homeostatic cues (low-energy intake and maintenance feeding) core body temperature tended to be higher in Calm compared with Nervous animals. In contrast, in response to nonhomeostatic cues, Nervous animals had higher anticipatory thermogenic responses than Calm animals. Expression of uncoupling protein (UCP)-1 and -2 mRNA were higher in retroperitoneal tissue than in subcutaneous tissue, but UCP3 and leptin mRNA levels were similar at both sites; expression of these genes was similar in Nervous and Calm animals. There were no differences in stress responsiveness. We conclude that temperament differentially influences adipose thermogenesis and the regulation of core body temperature in responses to both homeostatic and nonhomeostatic stimuli.

  19. Imaging of the skin and subcutaneous tissue using classical and high-frequency ultrasonographies in anti-cellulite therapy.

    Science.gov (United States)

    Mlosek, Robert Krzysztof; Dębowska, Renata Maria; Lewandowski, Marcin; Malinowska, Sylwia; Nowicki, Andrzej; Eris, Irena

    2011-11-01

    The development of ultrasonography allowed for skin imaging used in dermatology and esthetic medicine. By means of classic and high-frequency ultrasonographies, changes within the dermis and subcutaneous tissue can be presented. The aim of this study was to show the possibilities of applying classic and high-frequency ultrasonographies in esthetic dermatology based on monitoring various types of anti-cellulite therapies. Sixty-one women with cellulite were assigned to two smaller groups. One group was using anti-cellulite cream and the second group was a placebo group. The ultrasound examination was carried out before the initiation and after the completion of the treatment and evaluated epidermal echoes, the thickness of the subcutaneous tissue and the dermis, dermis echogenicity, the length and surface area of the subcutaneous tissue fascicles growing into the dermis, and the presence or absence of edemas. After the completion of the treatment, a statistically significant difference was observed. The most useful parameters were as follows: the thickness of the subcutaneous tissue, echogenicity, the surface area and length of the subcutaneous tissue, as well as the presence of edemas. The discussed changes were not observed in the placebo group. Classic and high-frequency ultrasonographies are useful methods for monitoring anti-cellulite therapies. © 2011 John Wiley & Sons A/S.

  20. Elucidation of the effects of a high fat diet on trace elements in rabbit tissues using atomic absorption spectroscopy.

    Science.gov (United States)

    Abdelhalim, Mohamed Anwar K; Alhadlaq, Hisham A; Moussa, Sherif Abdelmottaleb

    2010-01-12

    The mechanism of atherogenesis is not yet fully understood despite intense study in this area. The effects of high fat diet (HFD) on the changes of trace elements [iron (Fe), copper (Cu) and zinc (Zn)] in several tissues of rabbits have not been documented before. Thus, the aim of this study was to elucidate the changes in trace elements in several tissues of rabbits fed on HFD for a period of feeding of 10 weeks. The HFD group was fed a NOR rabbit chow supplemented with 1.0% cholesterol plus 1.0% olive oil. Fe, Cu and Zn concentrations were measured in four types of tissue from control and HFD rabbits using atomic absorption spectroscopy (AAS). Comparing HFD rabbits to control rabbits, we found that the highest percentage change of increase of Fe was 95% in lung tissue, while the lowest percentage change of increase of Fe was 7% in kidney tissue; the highest percentage change of decrease of Cu was 16% in aortic tissue, while the lowest percentage change of decrease of Cu was 6% in kidney tissue; and the highest percentage change of decrease of Zn was 71% in kidney tissue, while the lowest percentage change of decrease of Zn was 8% in lung tissue. These results suggest that Fe plays a major role in atherogenesis; it may accelerate the process of atherosclerosis probably through the production of free radicals, deposition and absorption of intracellular and extracellular lipids in the intima, connective tissue formation, smooth muscle proliferation, lower matrix degradation capacity and increased plaque stability. Furthermore, inducing anemia in HFD rabbits may delay or inhibit the progression of atherosclerosis. Cu plays a minor role in atherogenesis and Cu supplements may inhibit the progression of atherogenesis, perhaps by reducing the migration of smooth muscle cells from the media to the intima. Zn plays a major role in atherogenesis and that it may act as an endogenous protective factor against atherosclerosis perhaps by reducing lesion Fe content

  1. Contribution of skeletal muscle and adipose tissue to adrenaline-induced thermogenesis in man

    DEFF Research Database (Denmark)

    Simonsen, L; Stallknecht, B; Bülow, J

    1993-01-01

    subcutaneous adipose tissue metabolism was investigated. In both series Fick's principle was applied. Intravenous infusion increased blood flow, glucose uptake and oxygen uptake in both skeletal muscle and adipose tissue. It is concluded that skeletal muscle contributes about 40% and adipose tissue about 5......Elevated plasma adrenaline is known to increase whole body energy expenditure. We studied the thermogenic effect and the effects on substrate utilization in man during infusion of adrenaline. Two series were performed: in one series skeletal muscle metabolism was investigated and in another series...

  2. The thermogenic effect of leptin is dependent on a distinct population of prolactin-releasing peptide neurons in the dorsomedial hypothalamus.

    Science.gov (United States)

    Dodd, Garron T; Worth, Amy A; Nunn, Nicolas; Korpal, Aaron K; Bechtold, David A; Allison, Margaret B; Myers, Martin G; Statnick, Michael A; Luckman, Simon M

    2014-10-07

    Leptin is a critical regulator of metabolism, which acts on brain receptors (Lepr) to reduce energy intake and increase energy expenditure. Some of the cellular pathways mediating leptin's anorectic actions are identified, but those mediating the thermogenic effects have proven more difficult to decipher. We define a population of neurons in the dorsomedial hypothalamic nucleus (DMH) containing the RFamide PrRP, which is activated by leptin. Disruption of Lepr selectively in these cells blocks thermogenic responses to leptin and causes obesity. A separate population of leptin-insensitive PrRP neurons in the brainstem is required, instead, for the satiating actions of the gut-derived hormone cholecystokinin (CCK). Global deletion of PrRP (in a loxSTOPlox-PrRP mouse) results in obesity and attenuated responses to leptin and CCK. Cre-recombinase-mediated reactivation of PrRP in brainstem rescues the anorectic actions of CCK, but reactivation in the hypothalamus is required to re-establish the thermogenic effect of leptin. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  3. High intensity interval training improves liver and adipose tissue insulin sensitivity

    Directory of Open Access Journals (Sweden)

    Katarina Marcinko

    2015-12-01

    Conclusions: These data indicate that HIIT lowers blood glucose levels by improving adipose and liver insulin sensitivity independently of changes in adiposity, adipose tissue inflammation, liver lipid content or AMPK phosphorylation of ACC.

  4. Toward high-speed 3D nonlinear soft tissue deformation simulations using Abaqus software.

    Science.gov (United States)

    Idkaidek, Ashraf; Jasiuk, Iwona

    2015-12-01

    We aim to achieve a fast and accurate three-dimensional (3D) simulation of a porcine liver deformation under a surgical tool pressure using the commercial finite element software Abaqus. The liver geometry is obtained using magnetic resonance imaging, and a nonlinear constitutive law is employed to capture large deformations of the tissue. Effects of implicit versus explicit analysis schemes, element type, and mesh density on computation time are studied. We find that Abaqus explicit and implicit solvers are capable of simulating nonlinear soft tissue deformations accurately using first-order tetrahedral elements in a relatively short time by optimizing the element size. This study provides new insights and guidance on accurate and relatively fast nonlinear soft tissue simulations. Such simulations can provide force feedback during robotic surgery and allow visualization of tissue deformations for surgery planning and training of surgical residents.

  5. High quality tissue miniarray technique using a conventional TV/radio telescopic antenna.

    Science.gov (United States)

    Elkablawy, Mohamed A; Albasri, Abdulkader M

    2015-01-01

    The tissue microarray (TMA) is widely accepted as a fast and cost-effective research tool for in situ tissue analysis in modern pathology. However, the current automated and manual TMA techniques have some drawbacks restricting their productivity. Our study aimed to introduce an improved manual tissue miniarray (TmA) technique that is simple and readily applicable to a broad range of tissue samples. In this study, a conventional TV/radio telescopic antenna was used to punch tissue cores manually from donor paraffin embedded tissue blocks which were pre-incubated at 40oC. The cores were manually transferred, organized and attached to a standard block mould, and filled with liquid paraffin to construct TmA blocks without any use of recipient paraffin blocks. By using a conventional TV/radio antenna, it was possible to construct TmA paraffin blocks with variable formats of array size and number (2-mm x 42, 2.5-mm x 30, 3-mm x 24, 4-mm x 20 and 5-mm x 12 cores). Up to 2-mm x 84 cores could be mounted and stained on a standard microscopic slide by cutting two sections from two different blocks and mounting them beside each other. The technique was simple and caused minimal damage to the donor blocks. H and E and immunostained slides showed well-defined tissue morphology and array configuration. This technique is easy to reproduce, quick, inexpensive and creates uniform blocks with abundant tissues without specialized equipment. It was found to improve the stability of the cores within the paraffin block and facilitated no losses during cutting and immunostaining.

  6. Perceived Strengths and Weaknesses of Highly Realistic Training and Live Tissue Training for Navy Corpsmen

    Science.gov (United States)

    2015-04-08

    combat settings. Live tissue training involves the use of live specimens (typically pigs or goats) to practice specific medical procedures. The...trauma care training used by the military. In live tissue training, specimens (typically pigs or goats) are anesthetized, monitored, and finally...such as hemorrhage control and airway intubation, and provides immediate feedback about the quality of their care through their specimens’ vital signs

  7. Browning and graying: novel transcriptional regulators of brown and beige fat tissues and aging

    Directory of Open Access Journals (Sweden)

    Elisabetta eMueller

    2016-03-01

    Full Text Available Obesity represents a major risk factor for the development of a number of metabolic disorders, including cardiovascular disease and type 2 diabetes. Since the discovery that brown and beige fat cells exist in adult humans and contribute to energy expenditure, increasing interest has been devoted to the understanding of the molecular switches turning on calorie utilization. It has been reported that the ability of thermogenic tissues to burn energy declines during aging, possibly contributing to the development of metabolic dysfunction late in life. This review will focus on the recently identified transcriptional modulators of brown and beige cells and will discuss the potential impact of some of these thermogenic factors on age-associated metabolic disorders.

  8. A High-Dimensional Atlas of Human T Cell Diversity Reveals Tissue-Specific Trafficking and Cytokine Signatures.

    Science.gov (United States)

    Wong, Michael Thomas; Ong, David Eng Hui; Lim, Frances Sheau Huei; Teng, Karen Wei Weng; McGovern, Naomi; Narayanan, Sriram; Ho, Wen Qi; Cerny, Daniela; Tan, Henry Kun Kiaang; Anicete, Rosslyn; Tan, Bien Keem; Lim, Tony Kiat Hon; Chan, Chung Yip; Cheow, Peng Chung; Lee, Ser Yee; Takano, Angela; Tan, Eng-Huat; Tam, John Kit Chung; Tan, Ern Yu; Chan, Jerry Kok Yen; Fink, Katja; Bertoletti, Antonio; Ginhoux, Florent; Curotto de Lafaille, Maria Alicia; Newell, Evan William

    2016-08-16

    Depending on the tissue microenvironment, T cells can differentiate into highly diverse subsets expressing unique trafficking receptors and cytokines. Studies of human lymphocytes have primarily focused on a limited number of parameters in blood, representing an incomplete view of the human immune system. Here, we have utilized mass cytometry to simultaneously analyze T cell trafficking and functional markers across eight different human tissues, including blood, lymphoid, and non-lymphoid tissues. These data have revealed that combinatorial expression of trafficking receptors and cytokines better defines tissue specificity. Notably, we identified numerous T helper cell subsets with overlapping cytokine expression, but only specific cytokine combinations are secreted regardless of tissue type. This indicates that T cell lineages defined in mouse models cannot be clearly distinguished in humans. Overall, our data uncover a plethora of tissue immune signatures and provide a systemic map of how T cell phenotypes are altered throughout the human body. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. The Novel Gene CRNDE Encodes a Nuclear Peptide (CRNDEP Which Is Overexpressed in Highly Proliferating Tissues.

    Directory of Open Access Journals (Sweden)

    Lukasz Michal Szafron

    Full Text Available CRNDE, recently described as the lncRNA-coding gene, is overexpressed at RNA level in human malignancies. Its role in gametogenesis, cellular differentiation and pluripotency has been suggested as well. Herein, we aimed to verify our hypothesis that the CRNDE gene may encode a protein product, CRNDEP. By using bioinformatics methods, we identified the 84-amino acid ORF encoded by one of two CRNDE transcripts, previously described by our research team. This ORF was cloned into two expression vectors, subsequently utilized in localization studies in HeLa cells. We also developed a polyclonal antibody against CRNDEP. Its specificity was confirmed in immunohistochemical, cellular localization, Western blot and immunoprecipitation experiments, as well as by showing a statistically significant decrease of endogenous CRNDEP expression in the cells with transient shRNA-mediated knockdown of CRNDE. Endogenous CRNDEP localizes predominantly to the nucleus and its expression seems to be elevated in highly proliferating tissues, like the parabasal layer of the squamous epithelium, intestinal crypts or spermatocytes. After its artificial overexpression in HeLa cells, in a fusion with either the EGFP or DsRed Monomer fluorescent tag, CRNDEP seems to stimulate the formation of stress granules and localize to them. Although the exact role of CRNDEP is unknown, our preliminary results suggest that it may be involved in the regulation of the cell proliferation. Possibly, CRNDEP also participates in oxygen metabolism, considering our in silico results, and the correlation between its enforced overexpression and the formation of stress granules. This is the first report showing the existence of a peptide encoded by the CRNDE gene.

  10. Maternal high-fat diet exaggerates atherosclerosis in adult offspring by augmenting periaortic adipose tissue-specific proinflammatory response.

    Science.gov (United States)

    Wakana, Noriyuki; Irie, Daisuke; Kikai, Masakazu; Terada, Kensuke; Yamamoto, Keita; Kawahito, Hiroyuki; Kato, Taku; Ogata, Takehiro; Ueyama, Tomomi; Matoba, Satoaki; Yamada, Hiroyuki

    2015-03-01

    Maternal obesity elicits offspring's metabolic disorders via developmental modifications of visceral adipose tissue; however, its effect on atherogenesis remains undefined. Perivascular adipose tissue has recently been implicated in vascular remodeling and vasoreactivity. We hypothesize that developmental modifications of perivascular adipose tissue by maternal high-fat diet (HFD) exposure promotes atherosclerosis in adult offspring. Eight-week-old female apolipoprotein E-deficient mice were fed an HFD or normal diet (ND) during gestation and lactation. Offspring were fed a high-cholesterol diet from 8 weeks of age. Twenty-week-old male offspring of HFD-fed dams (O-HFD) showed a 2.1-fold increase in atherosclerotic lesion of the entire aorta compared with those of ND-fed dams (O-ND). Although mRNA expressions of interleukin-6, tumor necrosis factor, and monocyte chemotactic protein-1 and accumulation of macrophages in epididymal white adipose tissue were less in O-HFD than in O-ND, thoracic periaortic adipose tissue (tPAT) showed an exaggerated inflammatory response in O-HFD. Intra-abdominal transplantation of tPAT from 8-week-old O-HFD alongside the distal abdominal aorta exaggerated atherosclerosis development of the infrarenal aorta in recipient apolipoprotein E-deficient mice compared with tPAT from O-ND (210%, Patherosclerosis development in offspring by augmenting tPAT-specific inflammatory response proceeded by an increased expression of macrophage colony-stimulating factor. © 2015 American Heart Association, Inc.

  11. Label-free profiling of white adipose tissue of rats exhibiting high or low levels of intrinsic exercise capacity.

    Science.gov (United States)

    Bowden-Davies, Kelly; Connolly, Joanne; Burghardt, Paul; Koch, Lauren G; Britton, Steven L; Burniston, Jatin G

    2015-07-01

    Divergent selection has created rat phenotypes of high- and low-capacity runners (HCR and LCR, respectively) that have differences in aerobic capacity and correlated traits such as adiposity. We analyzed visceral adipose tissue of HCR and LCR using label-free high-definition MS (elevated energy) profiling. The running capacity of HCR was ninefold greater than LCR. Proteome profiling encompassed 448 proteins and detected 30 significant (p <0.05; false discovery rate <10%, calculated using q-values) differences. Approximately half of the proteins analyzed were of mitochondrial origin, but there were no significant differences in the abundance of proteins involved in aerobic metabolism. Instead, adipose tissue of LCR rats exhibited greater abundances of proteins associated with adipogenesis (e.g. cathepsin D), ER stress (e.g. 78 kDa glucose response protein), and inflammation (e.g. Ig gamma-2B chain C region). Whereas the abundance antioxidant enzymes such as superoxide dismutase [Cu-Zn] was greater in HCR tissue. Putative adipokines were also detected, in particular protein S100-B, was 431% more abundant in LCR adipose tissue. These findings reveal low running capacity is associated with a pathological profile in visceral adipose tissue proteome despite no detectable differences in mitochondrial protein abundance. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Novel Model-Based Inquiry of Ionic Bonding in Alginate Hydrogels Used in Tissue Engineering for High School Students

    Science.gov (United States)

    Bowles, Robby D.; Saroka, James M.; Archer, Shivaun D.; Bonassar, Lawrence J.

    2012-01-01

    Because of cost and time, it is difficult to relate to students how fundamental chemical principles are involved in cutting edge biomedical breakthroughs being reported in the national media. The laboratory exercise presented here is aimed at high school chemistry students and uses alginate hydrogels, a common material used in tissue engineering,…

  13. Comparison between high-frequency ultrasonography and histological assessment reveals weak correlation for measurements of scar tissue thickness.

    Science.gov (United States)

    Agabalyan, Natacha A; Su, Samuel; Sinha, Sarthak; Gabriel, Vincent

    2017-05-01

    Current methods for evaluating scar tissue volume following burns have shortcomings. The Vancouver Burn Scar scale is subjective, leading to a high variability in assessment. Although histological assessment via punch biopsy can discriminate between the different layers of skin, such an approach is invasive, inefficient, and detrimental to patient experience and wound healing. This study investigates the accuracy of high-frequency ultrasonography, a non-invasive alternative to histology, for measuring dermal and epidermal thickness in scar tissue. Scar thicknesses of 10 patients following burns were assessed using a 2-D high-frequency ultrasound probe. The scars were then biopsied using a circular 4mm punch biopsy for histological assessment. Dermal, epidermal, and total thickness of the scar tissue was measured using ultrasound and histology, and correlations between the two measurements were calculated. There was not a strong correlation between ultrasound measurement and histological analysis for epidermal, dermal, and total thickness (Spearman's rank correlation of -0.1223, -0.6242, and -0.6242) of scar tissue. Measurements of scar thickness using high-frequency ultrasonography did not recapitulate the in vivo dermal, epidermal and total thickness. Based on these findings, strategies for further optimization of 2-D ultrasonography is discussed before clinical and research use. Copyright © 2016 Elsevier Ltd and ISBI. All rights reserved.

  14. Practices and Procedures to Prevent the Transmission of Skin and Soft Tissue Infections in High School Athletes

    Science.gov (United States)

    Fritz, Stephanie A.; Long, Marcus; Gaebelein, Claude J.; Martin, Madeline S.; Hogan, Patrick G.; Yetter, John

    2012-01-01

    Skin and soft tissue infections (SSTIs) are frequent in student athletes and are often caused by community-associated methicillin-resistant "Staphylococcus aureus" (CA-MRSA). We evaluated the awareness of CA-MRSA among high school coaches and athletic directors in Missouri (n = 4,408) and evaluated hygiene practices affecting SSTI…

  15. Optimizing preservation protocols to extract high-quality RNA from different tissues of echinoderms for next-generation sequencing.

    Science.gov (United States)

    Pérez-Portela, Rocío; Riesgo, Ana

    2013-09-01

    Transcriptomic information provides fundamental insights into biological processes. Extraction of quality RNA is a challenging step, and preservation and extraction protocols need to be adjusted in many cases. Our objectives were to optimize preservation protocols for isolation of high-quality RNA from diverse echinoderm tissues and to compare the utility of parameters as absorbance ratios and RIN values to assess RNA quality. Three different tissues (gonad, oesophagus and coelomocytes) were selected from the sea urchin Arbacia lixula. Solid tissues were flash-frozen and stored at -80 °C until processed. Four preservation treatments were applied to coelomocytes: flash freezing and storage at -80 °C, RNAlater and storage at -20 °C, preservation in TRIzol reagent and storage at -80 °C and direct extraction with TRIzol from fresh cells. Extractions of total RNA were performed with a modified TRIzol protocol for all tissues. Our results showed high values of RNA quantity and quality for all tissues, showing nonsignificant differences among them. However, while flash freezing was effective for solid tissues, it was inadequate for coelomocytes because of the low quality of the RNA extractions. Coelomocytes preserved in RNAlater displayed large variability in RNA integrity and insufficient RNA amount for further isolation of mRNA. TRIzol was the most efficient system for stabilizing RNA which resulted on high RNA quality and quantity. We did not detect correlation between absorbance ratios and RNA integrity. The best strategies for assessing RNA integrity was the visualization of 18S rRNA and 28S rRNA bands in agarose gels and estimation of RIN values with Agilent Bioanalyzer chips. © 2013 John Wiley & Sons Ltd.

  16. Acute effects of ingesting a commercial thermogenic drink on changes in energy expenditure and markers of lipolysis

    Directory of Open Access Journals (Sweden)

    Stout Jeffrey R

    2008-02-01

    Full Text Available Abstract Background To determine the acute effects of ingesting a thermogenic drink (Celsius, Delray Beach, FL (TD on changes in metabolism and lipolysis. Methods Healthy college-aged male (23.2 ± 4.0 y, 177.2 ± 6.1 cm, 81.7 ± 11.3 kg, 22.8 ± 7.3 % fat; n = 30 and female (23.4 ± 3.1 y, 165.6 ± 8.7 cm, 62.1 ± 9.9 kg, 28.3 ± 7.4 % fat; n = 30 participants were matched according to height and weight to consume 336 ml of the TD or a non-caloric, non-caffeinated placebo (PLA. After a 12 h fast, participants reported for pre-consumption measures of height, weight, heart rate, blood pressure, resting energy expenditure (REE, respiratory exchange ratio (RER, glycerol and free-fatty acid (FFA concentrations. REE and RER were determined at 60, 120, and 180 min post-consumption. Serum glycerol and FFA concentrations were determined at 30, 60, 120 and 180 min post-consumption. Results When compared to PLA, TD significantly increased REE at 60, 120 and 180 min (p Conclusion Acute TD ingestion significantly increased REE, FFA and glycerol appearance. If sustained, these changes may help to promote weight loss and improve body composition; however, these findings are currently unknown as are the general safety and efficacy of prolonged consumption.

  17. The High Frequency Ultrasonic Diagnostic System for Hard and Soft Tissue Specific Assessments in Dentistry

    Science.gov (United States)

    Slak, Bartosz

    The numerical assessment of dental tissues is essential when selecting a relevant treatment protocol in the field of dentistry. This will have significant ramifications on the restoration quality of dental tissues. The aim of the research study presented in this thesis was to validate applicability and obtain non-invasively, quantitative data for hard and soft tissue thickness in dental applications. An ultrasonic system was developed and assembled for the purpose of these experiments. Numerous laboratory trials were conducted to validate system performance against traditional and destructive methods of assessment. Ultrasonic measurements were found to yield similar values to those obtained from invasive methods. Results obtained in these experiments have validated potentials of ultrasound as a supplementary diagnostic tool for dental healthcare.

  18. Microelement content of bone tissue in transosseous osteosynthesis by II-izarov technique in high-altitude destinations (experimental research

    OpenAIRE

    Erokhin A.N.; Isakov B.D.; Nakoskin A.N.

    2014-01-01

    Aim: To study the redistribution of microelements in bone tissue in osteosynthesis in high-altitude destinations. Material and Methods. The study was performed on 72 mongrel dogs of both sexes aged 1-3 years, weighing 10—15 kg. 32 dogs have been experienced the extended leg in lowlands destinations, another group of 32 animals have been experienced elongation produced in high-altitude destinations. The control group consisted of 8 healthy dogs of low mountain areas. The standard technique of ...

  19. Simultaneous isolation of high-quality DNA, RNA, miRNA and proteins from tissues for genomic applications.

    Science.gov (United States)

    Peña-Llopis, Samuel; Brugarolas, James

    2013-11-01

    Genomic technologies have revolutionized our understanding of complex Mendelian diseases and cancer. Solid tumors present several challenges for genomic analyses, such as tumor heterogeneity and tumor contamination with surrounding stroma and infiltrating lymphocytes. We developed a protocol to (i) select tissues of high cellular purity on the basis of histological analyses of immediately flanking sections and (ii) simultaneously extract genomic DNA (gDNA), mRNA, noncoding RNA (ncRNA; enriched in miRNA) and protein from the same tissues. After tissue selection, about 12-16 extractions of DNA, RNA or protein can be obtained per day. Compared with other similar approaches, this fast and reliable methodology allowed us to identify mutations in tumors with remarkable sensitivity and to perform integrative analyses of whole-genome and exome data sets, DNA copy numbers (by single-nucleotide polymorphism (SNP) arrays), gene expression data (by transcriptome profiling and quantitative PCR (qPCR)) and protein levels (by western blotting and immunohistochemical analysis) from the same samples. Although we focused on renal cell carcinoma, this protocol may be adapted with minor changes to any human or animal tissue to obtain high-quality and high-yield nucleic acids and proteins.

  20. Arterial and fat tissue inflammation are highly correlated: a prospective {sup 18}F-FDG PET/CT study

    Energy Technology Data Exchange (ETDEWEB)

    Bucerius, Jan [Mount Sinai School of Medicine, Translational and Molecular Imaging Institute, One Gustave L. Levy Place, P. O. Box 1234, New York, NY (United States); Mount Sinai School of Medicine, Department of Radiology, New York, NY (United States); Maastricht University Medical Center, Department of Nuclear Medicine, Maastricht (Netherlands); Maastricht University Medical Center, Cardiovascular Research Institute Maastricht (CARIM), Maastricht (Netherlands); University Hospital RWTH Aachen, Department of Nuclear Medicine, Aachen (Germany); Mani, Venkatesh; Fayad, Zahi A. [Mount Sinai School of Medicine, Translational and Molecular Imaging Institute, One Gustave L. Levy Place, P. O. Box 1234, New York, NY (United States); Mount Sinai School of Medicine, Department of Radiology, New York, NY (United States); Mount Sinai School of Medicine, Department of Cardiology, Zena and Michael A. Weiner Cardiovascular Institute and Marie-Josee and Henry R. Kravis Cardiovascular Health Center, New York, NY (United States); Wong, Stephanie; Moncrieff, Colin; Izquierdo-Garcia, David [Mount Sinai School of Medicine, Translational and Molecular Imaging Institute, One Gustave L. Levy Place, P. O. Box 1234, New York, NY (United States); Mount Sinai School of Medicine, Department of Radiology, New York, NY (United States); Machac, Josef [Mount Sinai School of Medicine, Division of Nuclear Medicine, Department of Radiology, New York, NY (United States); Fuster, Valentin [Mount Sinai School of Medicine, Department of Cardiology, Zena and Michael A. Weiner Cardiovascular Institute and Marie-Josee and Henry R. Kravis Cardiovascular Health Center, New York, NY (United States); The Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid (Spain); Farkouh, Michael E. [Mount Sinai School of Medicine, Department of Cardiology, Zena and Michael A. Weiner Cardiovascular Institute and Marie-Josee and Henry R. Kravis Cardiovascular Health Center, New York, NY (United States); Mount Sinai School of Medicine, Cardiovascular Imaging Clinical Trials Unit, New York, NY (United States); Rudd, James H.F. [University of Cambridge, Division of Cardiovascular Medicine, Cambridge (United Kingdom)

    2014-05-15

    There is evidence that the link between obesity and cardiovascular disease might relate to inflammation in both fat tissue and the arterial wall. {sup 18}F-FDG uptake on PET is a surrogate marker of vessel wall inflammation. The aim of the study was to measure FDG uptake in both regions using PET and identify links between adipose and arterial inflammation. Included in the study were 173 cardiovascular patients who were prospectively imaged with FDG PET/CT. Arterial FDG uptake was measured in the carotid arteries and ascending aorta. The same was done in fat tissue in the neck, the presternal region (both subcutaneous) and the pericardium. FDG uptake was quantified as average maximal target-to-background ratio ({sub mean}TBR{sub max}). Multivariate regression analyses were performed to identify significant associations between arterial and adipose tissue FDG uptake and clinical variables as given by the standardized correlation coefficient (β). FDG uptake values in all fat tissue regions were highly predictive of vascular FDG uptake in both the carotids (β 0.262, p < 0.0001, in the neck subcutaneous region) and aorta (β 0.22, p = 0.008, in the chest pericardial region; β 0.193, p = 0.019, in the chest subcutaneous region). Obesity was significantly associated with elevated FDG uptake in adipose tissue (β 0.470, p < 0.0001, in the neck subcutaneous region; β 0.619, p = 0.028, in the chest subcutaneous region; β 0.978, p = 0.035, in the chest pericardial region). FDG uptake in diverse fat tissue regions was significantly associated with arterial FDG uptake, a reasonable surrogate of inflammation. Increasing body weight significantly predicted the level of fatty inflammation. FDG PET therefore provides imaging evidence of an inflammatory link between fat tissue and the vasculature in patients with cardiovascular disease. (orig.)

  1. High Throughput Micro-Well Generation of Hepatocyte Micro-Aggregates for Tissue Engineering

    NARCIS (Netherlands)

    Gevaert, Elien; Dollé, Laurent; Billiet, Thomas; Dubruel, Peter; van Grunsven, Leo; van Apeldoorn, Aart A.; Cornelissen, Ria

    2014-01-01

    The main challenge in hepatic tissue engineering is the fast dedifferentiation of primary hepatocytes in vitro. One successful approach to maintain hepatocyte phenotype on the longer term is the cultivation of cells as aggregates. This paper demonstrates the use of an agarose micro-well chip for the

  2. MR thermometry in fat-containing tissues for MR-guided High Intensity Focused Ultrasound therapy

    NARCIS (Netherlands)

    Baron, Paul

    2015-01-01

    The aim of the research was to improve and apply MR thermometry techniques in fat -containing tissues. These thermometry methods were researched with the aim of applying them for MR-HIFU treatment of breast and liver tumors. The first study demonstrated the sensitivity of proton resonance frequency

  3. High-risk human papilloma virus in archival tissues of oral pathosis and normal oral mucosa

    Directory of Open Access Journals (Sweden)

    Raghu Dhanapal

    2015-01-01

    Full Text Available Objectives: Oral cancer ranks third among all cancers in the Indian population. Human papilloma virus (HPV plays a significant role in oral carcinogenesis. Population-based subtype variations are present in the HPV prevalence. This study gives an emphasis on the parameters to be considered in formalin fixed paraffin embedded tissues for polymerase chain reaction (PCR-based research work. Materials and Methods: Cross-sectional study on archival paraffin-embedded tissue samples of oral squamous cell carcinoma (OSCC, epithelial dysplasia, and normal oral mucosa surrounding impacted tooth was amplified by PCR for the E6 gene of HPV type 16 and E1 gene of HPV type 18. Results: HPV 18 was positive in three OSCC cases. There was no statistically significant association of the positivity of HPV with the age, gender or habit. The HPV positive patients had a tobacco habit and were of a younger age group. Conclusion: The presence of HPV in carcinomatous tissue highlights the possible role of HPV in carcinogenesis and archival paraffin embedded tissue specimen can be used for this analysis. Recent studies on genomic analyses have highlighted that the HPV positive tumors are a separate subgroup based on genomic sequencing. The results of a larger retrospective study will help further in our understanding of the role of HPV in carcinogenesis, this study could form the baseline for such follow-up studies.

  4. Short term, high fat-feeding induced changes in white adipose tissue gene expression are highly predictive for long term changes

    NARCIS (Netherlands)

    Voigt, A.; Agnew, K.; Schothorst, van E.M.; Keijer, J.; Klaus, S.

    2013-01-01

    Scope - We aimed to evaluate the predictability of short-term (5 days) changes in epididymal white adipose tissue (eWAT) gene expression for long-term (12 weeks) changes induced by high-fat diet (HFD) feeding. Methods and results - Mice were fed semisynthetic diets containing 10 (low-fat diet) or 40

  5. High-definition Fourier Transform Infrared (FT-IR) spectroscopic imaging of human tissue sections towards improving pathology.

    Science.gov (United States)

    Sreedhar, Hari; Varma, Vishal K; Nguyen, Peter L; Davidson, Bennett; Akkina, Sanjeev; Guzman, Grace; Setty, Suman; Kajdacsy-Balla, Andre; Walsh, Michael J

    2015-01-21

    High-definition Fourier Transform Infrared (FT-IR) spectroscopic imaging is an emerging approach to obtain detailed images that have associated biochemical information. FT-IR imaging of tissue is based on the principle that different regions of the mid-infrared are absorbed by different chemical bonds (e.g., C=O, C-H, N-H) within cells or tissue that can then be related to the presence and composition of biomolecules (e.g., lipids, DNA, glycogen, protein, collagen). In an FT-IR image, every pixel within the image comprises an entire Infrared (IR) spectrum that can give information on the biochemical status of the cells that can then be exploited for cell-type or disease-type classification. In this paper, we show: how to obtain IR images from human tissues using an FT-IR system, how to modify existing instrumentation to allow for high-definition imaging capabilities, and how to visualize FT-IR images. We then present some applications of FT-IR for pathology using the liver and kidney as examples. FT-IR imaging holds exciting applications in providing a novel route to obtain biochemical information from cells and tissue in an entirely label-free non-perturbing route towards giving new insight into biomolecular changes as part of disease processes. Additionally, this biochemical information can potentially allow for objective and automated analysis of certain aspects of disease diagnosis.

  6. [Rosuvastatin improves insulin sensitivity in overweight rats induced by high fat diet. Role of SIRT1 in adipose tissue].

    Science.gov (United States)

    Valero-Muñoz, María; Martín-Fernández, Beatriz; Ballesteros, Sandra; Cachofeiro, Victoria; Lahera, Vicente; de Las Heras, Natalia

    2014-01-01

    To study the effects of rosuvastatin on insulin resistance in overweight rats induced by high fat diet, as well as potential mediators. We used male Wistar rats fed with a standard diet (CT) or high fat diet (33.5% fat) (HFD); half of the animals HFD were treated with rosuvastatin (15mg/kg/day) (HFD+Rosu) for 7 weeks. HFD rats showed increased body, epididymal and lumbar adipose tissue weights. Treatment with Rosu did not modify body weight or the weight of the adipose packages in HFD rat. Plasma glucose and insulin levels and HOMA index were higher in HFD rats, and rosuvastatin treatment reduced them. Leptin/adiponectin ratio in plasma and lumbar adipose tissue were higher in HDF rats, and were reduced by rosuvastatin. SIRT-1, PPAR-γ and GLUT-4 protein expression in lumbar adipose tissue were lower in HFD rats and Rosu normalized expression of the three mediators. Rosuvastatin ameliorates insulin sensitivity induced by HFD in rats. This effect is mediated by several mechanisms including reduction of leptin and enhancement of SIRT-1, PPAR-γ and GLUT-4 expression in white adipose tissue. SIRT1 could be considered a major mediator of the beneficial effects of rosuvastatin on insulin sensitivity in overweight rats induced by diet. Copyright © 2013 Sociedad Española de Arteriosclerosis. Published by Elsevier España. All rights reserved.

  7. Nonlinear ultrasound propagation through layered liquid and tissue-equivalent media: computational and experimental results at high frequency.

    Science.gov (United States)

    Williams, Ross; Cherin, Emmanuel; Lam, Toby Y J; Tavakkoli, Jahangir; Zemp, Roger J; Foster, F Stuart

    2006-11-21

    Nonlinear propagation has been demonstrated to have a significant impact on ultrasound imaging. An efficient computational algorithm is presented to simulate nonlinear ultrasound propagation through layered liquid and tissue-equivalent media. Results are compared with hydrophone measurements. This study was undertaken to investigate the role of nonlinear propagation in high frequency ultrasound micro-imaging. The acoustic field of a focused transducer (20 MHz centre frequency, f-number 2.5) was simulated for layered media consisting of water and tissue-mimicking phantom, for several wide-bandwidth source pulses. The simulation model accounted for the effects of diffraction, attenuation and nonlinearity, with transmission and refraction at layer boundaries. The parameter of nonlinearity, B/A, of the water and tissue-mimicking phantom were assumed to be 5.2 and 7.4, respectively. The experimentally measured phantom B/A value found using a finite-amplitude insert-substitution method was shown to be 7.4 +/- 0.6. Relative amounts of measured second and third harmonic pressures as a function of the fundamental pressures at the focus were in good agreement with simulations. Agreement within 3% was found between measurements and simulations of the beam widths of the fundamental and second harmonic signals following propagation through the tissue phantom. The results demonstrate significant nonlinear propagation effects for high frequency imaging beams.

  8. A magnetic resonance imaging study on changes in rat mandibular bone marrow and pulp tissue after high-dose irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Wan; Lee, Byung Do [Dept. of Oral and Maxillofacial Radiology and Wonkwang Dental Research Institute, College of Dentistry, Wonkwang University, Iksan (Korea, Republic of); Lee, Kang Kyoo [Dept. of Radiation Oncology, School of Medicine, Wonkwang University, Iksan (Korea, Republic of); Koh, Kwang Joon [Dept. of Oral and Maxillofacial Radiology, School of Dentistry and Institute of Oral Bioscience, Chonbuk National University, Jeonju (Korea, Republic of)

    2014-03-15

    This study was designed to evaluate whether magnetic resonance imaging (MRI) is appropriate for detecting early changes in the mandibular bone marrow and pulp tissue of rats after high-dose irradiation. The right mandibles of Sprague-Dawley rats were irradiated with 10 Gy (Group 1, n=5) and 20 Gy (Group 2, n=5). Five non-irradiated animals were used as controls. The MR images of rat mandibles were obtained before irradiation and once a week until week 4 after irradiation. From the MR images, the signal intensity (SI) of the mandibular bone marrow and pulp tissue of the incisor was interpreted. The MR images were compared with the histopathologic findings. The SI of the mandibular bone marrow had decreased on T2-weighted MR images. There was little difference between Groups 1 and 2. The SI of the irradiated groups appeared to be lower than that of the control group. The histopathologic findings showed that the trabecular bone in the irradiated group had increased. The SI of the irradiated pulp tissue had decreased on T2-weighted MR images. However, the SI of the MR images in Group 2 was high in the atrophic pulp of the incisor apex at week 2 after irradiation. These patterns seen on MRI in rat bone marrow and pulp tissue were consistent with histopathologic findings. They may be useful to assess radiogenic sclerotic changes in rat mandibular bone marrow.

  9. High-definition Fourier Transform Infrared (FT-IR) Spectroscopic Imaging of Human Tissue Sections towards Improving Pathology

    Science.gov (United States)

    Nguyen, Peter L.; Davidson, Bennett; Akkina, Sanjeev; Guzman, Grace; Setty, Suman; Kajdacsy-Balla, Andre; Walsh, Michael J.

    2015-01-01

    High-definition Fourier Transform Infrared (FT-IR) spectroscopic imaging is an emerging approach to obtain detailed images that have associated biochemical information. FT-IR imaging of tissue is based on the principle that different regions of the mid-infrared are absorbed by different chemical bonds (e.g., C=O, C-H, N-H) within cells or tissue that can then be related to the presence and composition of biomolecules (e.g., lipids, DNA, glycogen, protein, collagen). In an FT-IR image, every pixel within the image comprises an entire Infrared (IR) spectrum that can give information on the biochemical status of the cells that can then be exploited for cell-type or disease-type classification. In this paper, we show: how to obtain IR images from human tissues using an FT-IR system, how to modify existing instrumentation to allow for high-definition imaging capabilities, and how to visualize FT-IR images. We then present some applications of FT-IR for pathology using the liver and kidney as examples. FT-IR imaging holds exciting applications in providing a novel route to obtain biochemical information from cells and tissue in an entirely label-free non-perturbing route towards giving new insight into biomolecular changes as part of disease processes. Additionally, this biochemical information can potentially allow for objective and automated analysis of certain aspects of disease diagnosis. PMID:25650759

  10. A high-throughput, homogeneous microplate assay for agents that kill mammalian tissue culture cells.

    Science.gov (United States)

    Pierce, Michael; Wang, Chunwei; Rebentisch, Matt; Endo, Mark; Stump, Mark; Kamb, Alexander

    2003-06-01

    Screens for cytostasis/cytoxicity have considerable value for the discovery of therapeutic agents and the investigation of the biology of apoptosis. For instance, genetic screens for proteins, protein fragments, peptides, RNAs, or chemicals that kill tissue culture cells may aid in identifying new cancer therapeutic targets. A microplate assay for cell death is needed to achieve throughputs sufficient to sift through thousands of agents from expression or chemical libraries. The authors describe a homogeneous assay for cell death in tissue culture cells compatible with 96- or 384-well plates. In combination with a previously described system for retroviral packaging and transduction, nearly 6000 expression library clones could be screened per week in a 96-well plate format. The screening system may also prove useful for chemical screens.

  11. A High-Strength, Absorbable, Antibacterial Knotless Tissue Control Device for Fascial Closure

    OpenAIRE

    Nawrocki, Jesse G.; Nonnenmann, Heather; Mooney, Mark; Sutton, Nadia; Schmitz, Niels-Derrek

    2017-01-01

    Purpose of Review This review provides an overview of the STRATAFIX? SYMMETRIC PDS? Plus Knotless Tissue Control Device design and performance characteristics and highlights the device?s relevance for use in gynecological procedures. Various device testing was conducted on tensile strength, fixation tab mass comparison to conventional suture knot tower, initiation stitch strength, and wound holding strength to highlight the STRATAFIX? SYMMETRIC PDS? Plus Device?s key product attributes that m...

  12. Gene expression identifies heterogeneity of metastatic propensity in high-grade soft tissue sarcomas

    DEFF Research Database (Denmark)

    Skubitz, Keith M; Francis, Princy; Skubitz, Amy P N

    2012-01-01

    Metastatic propensity of soft tissue sarcoma (STS) is heterogeneous and may be determined by gene expression patterns that do not correlate well with morphology. The authors have reported gene expression patterns that distinguish 2 broad classes of clear cell renal carcinoma (ccRCC-gene set......), and other patterns that can distinguish heterogeneity of serous ovarian carcinoma (OVCA-gene set) and aggressive fibromatosis (AF-gene set); however, clinical follow-up data were not available for these samples....

  13. Highly purified collagen coating enhances tissue adherence and integration properties of monofilament polypropylene meshes.

    Science.gov (United States)

    Siniscalchi, Rodrigo Teixeira; Melo, Marli; Palma, Paulo César Rodrigues; Dal Fabbro, Inácio Maria; Vidal, Benedicto de Campos; Riccetto, Cassio Luiz Zanettini

    2013-10-01

    Complications related to tissue integration of polypropylene implants used in the treatment of pelvic organ prolapse are relatively prevalent. Collagen, a biocompatible, less immunogenic material with modulating properties on the inflammatory process, may improve polypropylene integration. The objective was to study biomechanical and histological effects of monofilament polypropylene mesh coated with purified collagen gel. Forty rats were implanted with two fragments of polypropylene mesh in their abdominal walls (one on each side of the linea alba). One of the fragments had a collagen gel coating (group I) while the other one did not (group II). The animals were euthanized at 7, 14, 90, and 180 days after implantation and their abdominal walls were excised for analysis. The biomechanical study showed that mesh adherence to neighboring tissue increased significantly in group II (p Polypropylene mesh coated with purified collagen gel increases adherence to tissue, promotes a less intense and lasting inflammatory response and triggers a greater organization and packing arrangement of collagen fibers in the late phase of implantation.

  14. An injection molding process for manufacturing highly porous and interconnected biodegradable polymer matrices for use as tissue engineering scaffolds.

    Science.gov (United States)

    Kramschuster, Adam; Turng, Lih-Sheng

    2010-02-01

    In this research, injection molding was combined with a novel material combination, supercritical fluid processing, and particulate leaching techniques to produce highly porous and interconnected structures that have the potential to act as scaffolds for tissue engineering applications. The foamed structures, molded with polylactide (PLA) and polyvinyl alcohol (PVOH) with salt as the particulate, were processed without the aid of organic solvents, which can be detrimental to tissue growth. The pore size in the scaffolds is controlled by salt particulates and interconnectivity is achieved by the co-continuous blending morphology of biodegradable PLA matrix with water-soluble PVOH. Carbon dioxide (CO(2)) at the supercritical state is used to serve as a plasticizer, thereby imparting moldability of blends even with an ultra high salt particulate content, and allows the use of low processing temperatures, which are desirable for temperature-sensitive biodegradable polymers. Interconnected pores of approximately 200 microm in diameter and porosities of approximately 75% are reported and discussed.

  15. Seroma suppression using TissuGlu® in a high-risk patient post-mastectomy: a case report.

    Science.gov (United States)

    Eichler, Christian; Dahdouh, Faten; Sauerwald, Axel; Warm, Mathias

    2013-05-28

    Post-mastectomy seromas are a common problem in modern oncological surgery. Occurrence rates of up to 59% have been reported in the literature. High-risk patients, that is, those who have received previous surgeries, radiation or chemotherapy, present a particular challenge. Several surgical techniques, including progressive tension suture application, have shown promise. Noninvasive measures such as fibrin-based adhesives have thus far not been able to prevent seroma occurrence effectively. A recent study using a lysine-derived urethane adhesive named TissuGlu®, however, showed promising results in patients after abdominoplasty. We used TissuGlu® to treat a high-risk 64-year-old female patient with a history of breast cancer and severe post-mastectomy seroma. The postsurgical period showed successful seroma suppression, without any adverse effects or complications. This type of adhesive should be evaluated as an alternative, less-invasive option for preventing seroma in patients after a mastectomy.

  16. β-Type Zr-Nb-Ti biomedical materials with high plasticity and low modulus for hard tissue replacements.

    Science.gov (United States)

    Nie, Li; Zhan, Yongzhong; Hu, Tong; Chen, Xiaoxian; Wang, Chenghui

    2014-01-01

    In order to develop new biomedical materials for hard tissue replacements, Zr-20Nb-xTi (x=0, 3, 7, 11 and 15) alloys with required properties were designed and prepared by using the vacuum arc melting method for the first time. Phase analysis and microstructural observation showed that all the as cast samples consisted of equiaxed β-Zr phase. The mechanical properties and fracture behaviors of the Zr-20Nb-xTi alloys have been analyzed. It is found that these alloys exhibit high plasticity, moderate compressive strength (1044-1325MPa) and yield stress (854-1080MPa), high elastic energy (12-20MJ/m(3)) and low Young's modulus (28-31GPa). This good combination of mechanical properties makes them potential biomedical materials for hard tissue replacement. © 2013 Elsevier Ltd. All rights reserved.

  17. No-cost manual method for preparation of tissue microarrays having high quality comparable to semiautomated methods.

    Science.gov (United States)

    Foda, Abd Al-Rahman Mohammad

    2013-05-01

    Manual tissue microarray (TMA) construction had been introduced to avoid the high cost of automated and semiautomated techniques. The cheapest and simplest technique for constructing manual TMA was that of using mechanical pencil tips. This study was carried out to modify this method, aiming to raise its quality to reach that of expensive ones. Some modifications were introduced to Shebl's technique. Two conventional mechanical pencil tips of different diameters were used to construct the recipient blocks. A source of mild heat was used, and blocks were incubated at 38°C overnight. With our modifications, 3 high-density TMA blocks were constructed. We successfully performed immunostaining without substantial tissue loss. Our modifications increased the number of cores per block and improved the stability of the cores within the paraffin block. This new, modified technique is a good alternative for expensive machines in many laboratories.

  18. Effect of hydrodynamic cavitation in the tissue erosion by pulsed high-intensity focused ultrasound (pHIFU)

    Science.gov (United States)

    Zhou, Yufeng; Gao, Xiaobin Wilson

    2016-09-01

    High-intensity focused ultrasound (HIFU) is emerging as an effective therapeutic modality in clinics. Besides the thermal ablation, tissue disintegration is also possible because of the interaction between the distorted HIFU bursts and either bubble cloud or boiling bubble. Hydrodynamic cavitation is another type of cavitation and has been employed widely in industry, but its role in mechanical erosion to tissue is not clearly known. In this study, the bubble dynamics immediately after the termination of HIFU exposure in the transparent gel phantom was captured by high-speed photography, from which the bubble displacement towards the transducer and the changes of bubble size was quantitatively determined. The characteristics of hydrodynamic cavitation due to the release of the acoustic radiation force and relaxation of compressed surrounding medium were found to associate with the number of pulses delivered and HIFU parameters (i.e. pulse duration and pulse repetition frequency). Because of the initial big bubble (~1 mm), large bubble expansion (up to 1.76 folds), and quick bubble motion (up to ~1 m s-1) hydrodynamic cavitation is significant after HIFU exposure and may lead to mechanical erosion. The shielding effect of residual tiny bubbles would reduce the acoustic energy delivered to the pre-existing bubble at the focus and, subsequently, the hydrodynamic cavitation effect. Tadpole shape of mechanical erosion in ex vivo porcine kidney samples was similar to the contour of bubble dynamics in the gel. Liquefied tissue was observed to emit towards the transducer through the punctured tissue after HIFU exposure in the sonography. In summary, the release of HIFU exposure-induced hydrodynamic cavitation produces significant bubble expansion and motion, which may be another important mechanism of tissue erosion. Understanding its mechanism and optimizing the outcome would broaden and enhance HIFU applications.

  19. Effect of hydrodynamic cavitation in the tissue erosion by pulsed high-intensity focused ultrasound (pHIFU).

    Science.gov (United States)

    Zhou, Yufeng; Gao, Xiaobin Wilson

    2016-09-21

    High-intensity focused ultrasound (HIFU) is emerging as an effective therapeutic modality in clinics. Besides the thermal ablation, tissue disintegration is also possible because of the interaction between the distorted HIFU bursts and either bubble cloud or boiling bubble. Hydrodynamic cavitation is another type of cavitation and has been employed widely in industry, but its role in mechanical erosion to tissue is not clearly known. In this study, the bubble dynamics immediately after the termination of HIFU exposure in the transparent gel phantom was captured by high-speed photography, from which the bubble displacement towards the transducer and the changes of bubble size was quantitatively determined. The characteristics of hydrodynamic cavitation due to the release of the acoustic radiation force and relaxation of compressed surrounding medium were found to associate with the number of pulses delivered and HIFU parameters (i.e. pulse duration and pulse repetition frequency). Because of the initial big bubble (~1 mm), large bubble expansion (up to 1.76 folds), and quick bubble motion (up to ~1 m s-1) hydrodynamic cavitation is significant after HIFU exposure and may lead to mechanical erosion. The shielding effect of residual tiny bubbles would reduce the acoustic energy delivered to the pre-existing bubble at the focus and, subsequently, the hydrodynamic cavitation effect. Tadpole shape of mechanical erosion in ex vivo porcine kidney samples was similar to the contour of bubble dynamics in the gel. Liquefied tissue was observed to emit towards the transducer through the punctured tissue after HIFU exposure in the sonography. In summary, the release of HIFU exposure-induced hydrodynamic cavitation produces significant bubble expansion and motion, which may be another important mechanism of tissue erosion. Understanding its mechanism and optimizing the outcome would broaden and enhance HIFU applications.

  20. Fungal endophytes in above-ground tissues of desert plants: infrequent in culture, but highly diverse and distinctive symbionts

    OpenAIRE

    Massimo, Nicholas C.; Nandi Devan, MM; Arendt, Kayla R.; Wilch, Margaret H.; Riddle, Jakob M.; Furr, Susan H.; Steen, Cole; U?Ren, Jana M.; Sandberg, Dustin C.; Arnold, A. Elizabeth

    2015-01-01

    In hot deserts, plants cope with aridity, high temperatures, and nutrient-poor soils with morphological and biochemical adaptations that encompass intimate microbial symbioses. Whereas the root microbiomes of arid-land plants have received increasing attention, factors influencing assemblages of symbionts in above-ground tissues have not been evaluated for many woody plants that flourish in desert environments. We evaluated the diversity, host affiliations, and distributions of endophytic fun...

  1. Sca-1 expression identifies stem cells in the proximal region of prostatic ducts with high capacity to reconstitute prostatic tissue

    OpenAIRE

    Patricia E Burger; Xiong, Xiaozhong; Coetzee, Sandra; Salm, Sarah N.; Moscatelli, David; Goto, Ken; Wilson, E. Lynette

    2005-01-01

    We previously showed that prostatic stem cells are concentrated in the proximal regions of prostatic ducts. We now report that these stem cells can be purified from isolated proximal duct regions by virtue of their high expression of the cell surface protein stem cell antigen 1 (Sca-1). In an in vivo prostate reconstitution assay, the purified Sca-1-expressing cell population isolated from the proximal region of ducts was more effective in generating prostatic tissue than a comparable populat...

  2. [Comparison of thermogenic character of liver and muscle in Chinese bulbul Pycnonotus sinensis between summer and winter].

    Science.gov (United States)

    Zheng, Wei-Hong; Fang, Yuan-Yuan; Jiang, Xue-Hua; Zhang, Guo-Kai; Liu, Jin-Song

    2010-06-01

    Acclimatization to winter conditions is an essential prerequisite for survival of small passerines. Seasonal changes in a bird's physiology and behavior are considered to be part of an adaptive strategy for survival and reproductive success. Changes in photoperiod, ambient temperature and food availability trigger seasonal acclimatization in physiology and behavior of many birds. In the present study, seasonal adjustments in several physiological, hormonal, and biochemical markers were examined in wild-captured Chinese bulbuls (Pycnonotus sinensis) from the Zhejiang Province in China. Oxygen consumption was measured using the closed-circuit respirometer containing 3.6 L animal chambers. State-4 respiration in liver and muscle mitochondria was measured at 30 degree with a Clark electrode. The activities of cytochrome C oxidase (COX) in liver and muscle were measured polarographically at 30 degree using a Clark electrode. The protein content of mitochondria was determined by the Folin phenol method, with bovine serum albumin as standard. In winter sparrows had higher body mass and basal metabolic rate (BMR). The contents of mitochondrial protein in liver, and state-4 respiration and COX activity in liver and muscle increased significantly in winter. Circulating level of serum triiodothyronine (T3) was significantly higher in winter than in summer. Together, these data suggest that Chinese bulbuls mainly coped with cold by enhancing thermogenic capacities through increased activity of respiratory enzymes activities. The results support the view that prominent winter increases in BMR are manifestations of winter acclimatization in Chinese bulbuls and that seasonal variation in metabolism in bulbuls is similar to that in other small wintering birds.

  3. Differentiation of pre-existing trapped methane from thermogenic methane in an igneous-intruded coal by hydrous pyrolysis

    Science.gov (United States)

    Dias, Robert F.; Lewan, Michael D.; Birdwell, Justin E.; Kotarba, Maciej J.

    2014-01-01

    So as to better understand how the gas generation potential of coal changes with increasing rank, same-seam samples of bituminous coal from the Illinois Basin that were naturally matured to varying degrees by the intrusion of an igneous dike were subjected to hydrous pyrolysis (HP) conditions of 360 °C for 72 h. The accumulated methane in the reactor headspace was analyzed for δ13C and δ2H, and mol percent composition. Maximum methane production (9.7 mg/g TOC) occurred in the most immature samples (0.5 %Ro), waning to minimal methane values at 2.44 %Ro (0.67 mg/g TOC), and rebounding to 3.6 mg/g TOC methane in the most mature sample (6.76 %Ro). Methane from coal with the highest initial thermal maturity (6.76 %Ro) shows no isotopic dependence on the reactor water and has a microbial δ13C value of −61‰. However, methane from coal of minimal initial thermal maturity (0.5 %Ro) shows hydrogen isotopic dependence on the reaction water and has a δ13C value of −37‰. The gas released from coals under hydrous pyrolysis conditions represents a quantifiable mixture of ancient (270 Ma) methane (likely microbial) that was generated in situ and trapped within the rock during the rapid heating by the dike, and modern (laboratory) thermogenic methane that was generated from the indigenous organic matter due to thermal maturation induced by hydrous pyrolysis conditions. These findings provide an analytical framework for better assessment of natural gas sources and for differentiating generated gas from pre-existing trapped gas in coals of various ranks.

  4. Olive leaf extract from Sicilian cultivar reduced lipid accumulation by inducing thermogenic pathway during adipogenesis

    Directory of Open Access Journals (Sweden)

    Rosa ePalmeri

    2016-05-01

    Full Text Available Olive leaves contain a wide variety of phenolic compounds belonging to phenolic acids, phenolic alcohols, flavonoids and secoiridoids, and include also many other pharmacological active compounds. They could play an important role in human diet and health because of their ability to lower blood pressure, increase coronary arteries blood flow and decrease the risk of cardiovascular diseases. The aim of this study was to investigate the effect of Olive leaf extract (OLE from Sicilian cultivar on adipogenic differentiation of human adipose derived mesenchymal stem cells and its impact on lipid metabolism. We showed that OLE treatment during adipogenic differentiation reduces inflammation, lipid accumulation and induces thermogenesis by activation of uncoupling protein UCP1, SIRT-1, PPARα and Pgc-1α. Furthermore, OLE significantly decreases the expression of molecules involved in adipogenesis and upregulates the expression of mediators involved in thermogenesis and lipid metabolism. Taken together, our results suggest that OLE may promote the brown remodeling of white adipose tissue inducing thermogenesis and improving metabolic homeostasis.

  5. Frequent Hypermethylation of RASSF1A, TSLC1, High Viral Load of Epstein-Barr Virus DNA in Nasopharyngeal Carcinoma, Matched Tumor-Adjacent Tissues

    Directory of Open Access Journals (Sweden)

    Liang Zhou

    2005-09-01

    Full Text Available We examined the promoter hypermethylation of tumorsuppressor genes RASSF1A, TSLC1, quantitated EBV DNA load in nasopharyngeal carcinoma (NPC tissues (T tissues, matched tumor-adjacent tissues outside 0.5 cm (P tissues, outside 1.0 cm (Z tissues to evaluate the role of promoter hypermethylation of RASSF1A, TSLC1 as well as viral load in the pathogenesis of NPC. Methylation-specific polymerase chain reaction (PCR for RASSF1A, TSLC1, quantitative real-time PCR analysis of EBV DNA were performed on matched T, P, Z tissues (n = 28 as well as chronic nasopharyngitis tissues (n = 8. Hypermethylated RASSF1A was frequently detected in the T (82%, P tissues (75%, but less frequently in Z tissues (46%. The average quantities of EBV DNA (copies/μg DNA in matched T, P, Z tissues were 673,000, 90,000, 7000. The differences of promoter hypermethylation of RASSF1A, EBV viral load among T, P, Z tissues were statistically significant, with more frequent methylation, higher viral load detected when tissues examined were nearer to the NPC tissues. Our results suggest that aberrant hypermethylation of RASSF1A, high EBV load might be important events in NPC pathogenesis, they may be useful molecular diagnostic markers for this cancer.

  6. Coagulation and ablation patterns of high-intensity focused ultrasound on a tissue-mimicking phantom and cadaveric skin.

    Science.gov (United States)

    Kim, Hee-Jin; Kim, Han Gu; Zheng, Zhenlong; Park, Hyoun Jun; Yoon, Jeung Hyun; Oh, Wook; Lee, Cheol Woo; Cho, Sung Bin

    2015-12-01

    High-intensity focused ultrasound (HIFU) can be applied noninvasively to create focused zones of tissue coagulation on various skin layers. We performed a comparative study of HIFU, evaluating patterns of focused tissue coagulation and ablation upon application thereof. A tissue-mimicking (TM) phantom was prepared with bovine serum albumin and polyacrylamide hydrogel to evaluate the geometric patterns of HIFU-induced thermal injury zones (TIZs) for five different HIFU devices. Additionally, for each device, we investigated histologic patterns of HIFU-induced coagulation and ablation in serial sections of cadaveric skin of the face and neck. All HIFU devices generated remarkable TIZs in the TM phantom, with different geometric values of coagulation for each device. Most of the TIZs seemed to be separated into two or more tiny parts. In cadaveric skin, characteristic patterns of HIFU-induced ablation and coagulation were noted along the mid to lower dermis at the focal penetration depth of 3 mm and along subcutaneous fat to the superficial musculoaponeurotic system or the platysma muscle of the neck at 4.5 mm. Additionally, remarkable pre-focal areas of tissue coagulation were observed in the upper and mid dermis at the focal penetration depth of 3 mm and mid to lower dermis at 4.5 mm. For five HIFU devices, we outlined various patterns of HIFU-induced TIZ formation along pre-focal, focal, and post-focal areas of TM phantom and cadaveric skin of the face and neck.

  7. Sarcolipin knockout mice fed a high-fat diet exhibit altered indices of adipose tissue inflammation and remodeling.

    Science.gov (United States)

    MacPherson, Rebecca E K; Gamu, Daniel; Frendo-Cumbo, Scott; Castellani, Laura; Kwon, Frenk; Tupling, A Russell; Wright, David C

    2016-07-01

    To investigate indices of adipose tissue inflammation and remodeling in high-fat diet (HFD) sarcolipin-knockout (SLN(-) (/-) ) mice. SLN regulates muscle-based nonshivering thermogenesis and is up-regulated with HFD. SLN(-) (/-) mice develop greater diet-induced obesity and glucose intolerance. This is accompanied by increases in circulating catecholamines and fatty acids. Catecholamines and fatty acids play a role in the pathology of adipose tissue inflammation. Male mice (wild type and SLN(-) (/-) ) were fed a HFD (42% kcal from fat) for 8 weeks. SLN(-) (/-) mice displayed greater obesity and glucose intolerance. This was accompanied by higher circulating epinephrine and nonesterified fatty acids. Epididymal but not inguinal subcutaneous adipose tissue from SLN(-) (/-) mice displayed higher interleukin-6, suppressor of cytokine signaling 3, interleukin-1β, and tumor necrosis factor-α mRNA expression, and this was associated with increased markers of macrophage infiltration (F4/80 expression and crown-like structures) and M1 polarization (higher CD11c expression and CD11c/MGL1). Interestingly, this occurred despite SLN(-) (/-) mice having smaller adipocytes. In conditions of nutrient excess, SLN(-) (/-) mice display depot-specific increases in indices of adipose tissue inflammation and remodeling. This could be a compensatory response to reductions in muscle-based thermogenesis. © 2016 The Obesity Society.

  8. Lactobacillus gasseri SBT2055 inhibits adipose tissue inflammation and intestinal permeability in mice fed a high-fat diet.

    Science.gov (United States)

    Kawano, Michio; Miyoshi, Masaya; Ogawa, Akihiro; Sakai, Fumihiko; Kadooka, Yukio

    2016-01-01

    The probiotic Lactobacillus gasseri SBT2055 (LG2055) has anti-obesity effects. Obesity is closely correlated with inflammation in adipose tissue, and maintaining adipose tissue in a less-inflamed state requires intestinal integrity or a barrier function to protect the intestine from the disruption that can be caused by a high-fat diet (HFD). Here, we examined the anti-inflammatory and intestinal barrier-protecting effects of LG2055 in C57BL/6 mice fed a normal-fat diet (NFD), HFD, or the HFD containing LG2055 (HFD-LG) for 21 weeks. HFD-LG intake significantly prevented HFD-induced increases in body weight, visceral fat mass, and the ratio of inflammatory-type macrophages to anti-inflammatory ones in adipose tissue. Mice fed the HFD showed higher intestinal permeability to a fluorescent dextran administered by oral administration and an elevated concentration of antibodies specific to lipopolysaccharides (LPS) in the blood compared with those fed the NFD, suggesting an increased penetration of the gut contents into the systemic circulation. These elevations of intestinal permeability and anti-LPS antibody levels were significantly suppressed in mice fed the HFD-LG. Moreover, treatment with LG2055 cells suppressed an increase in the cytokine-induced permeability of Caco-2 cell monolayers. These results suggest that LG2055 improves the intestinal integrity, reducing the entry of inflammatory substances like LPS from the intestine, which may lead to decreased inflammation in adipose tissue.

  9. Differential effects of acute and chronic estrogen treatment on thermogenic and metabolic pathways in ovariectomized sheep.

    Science.gov (United States)

    Clarke, Scott D; Clarke, Iain J; Rao, Alexandra; Evans, Roger G; Henry, Belinda A

    2013-01-01

    Estrogen is protective against weight gain, but the underlying mechanisms are not fully elucidated. We sought to characterize the effects of estrogen on energy expenditure in skeletal muscle and adipose tissue in ovariectomized sheep. Temperature probes were implanted into sc (gluteal) and visceral (retroperitoneal) fat depots and skeletal muscle of the hind limb (vastus lateralis). Food was available from 1100-1600 h to entrain postprandial thermogenesis. We characterized the effects of single (50 μg estradiol benzoate, im) and repeated (25 μg estradiol-17β, iv) injections as well as chronic (3 × 3 cm estradiol-17β implants for 7 d) treatment on heat production. A single injection of estrogen increased heat production in visceral fat and skeletal muscle, without an effect on food intake. Increased heat production in skeletal muscle was sustained by repeated estradiol-17β injections. On the other hand, continuous treatment reduced food intake but had no effect on thermogenesis. To determine possible mechanisms that underpin estradiol-17β-induced heat production, we measured femoral artery blood flow, the expression of uncoupling protein (UCP) mRNA and the phosphorylation of AMP-activated protein kinase and Akt in fat and muscle. There was little effect of either single or repeated injections of estradiol-17β on the expression of UCP1, -2, or -3 mRNA in visceral fat or skeletal muscle. Acute injection of estradiol-17β increased the phosphorylation of AMP-activated protein kinase and Akt in muscle only. Estradiol-17β treatment did not alter femoral artery blood flow. Thus, the stimulatory effect of estradiol-17β on thermogenesis in female sheep is dependent upon a pulsatile pattern of treatment and not constant continuous exposure.

  10. ROS and Sympathetically mediated Mitochondria activation in Brown Adipose Tissue contributes to Methamphetamine-induced hyperthermia

    Directory of Open Access Journals (Sweden)

    Manuel eSanchez-Alavez

    2013-04-01

    Full Text Available Methamphetamine abuse has been shown to induce alterations in mitochondrial function in the brain as well as to induce hyperthermia, which contributes to neurotoxicity and Meth-associated mortality. Brown adipose tissue (BAT, a thermogenic site known to be important in neonates, has recently regained importance since being identified in significant amounts and in correlation with metabolic balance in human adults. Given the high mitochondrial content of BAT and its role in thermogenesis, we aimed to investigate whether BAT plays any role in the development of Meth-induced hyperthermia. By ablating or denervating BAT, we identified a partial contribution of this organ to Meth-induced hyperthermia. BAT ablation decreased temperature by 0.5oC and reduced the length of hyperthermia by 1 hr, compared to sham-operated controls. BAT denervation also affected the development of hyperthermia in correlation with decreased the expression of electron transport chain molecules, and increase on PCG1a levels, but without affecting Meth-induced UCP1 upregulation. Furthermore, in isolated BAT cells in culture, Meth, but not Norepinephrine (NE, induced H2O2 upregulation. In addition, we found that in vivo Reactive Oxygen Species (ROS play a role in Meth hyperthermia. Thus, sympathetically- mediated mitochondrial activation in the BAT and Meth-induced ROS are key components to the development of hyperthermia in Meth abuse.

  11. Hypothalamic regulation of brown adipose tissue thermogenesis and energy homeostasis

    Directory of Open Access Journals (Sweden)

    Wei eZhang

    2015-08-01

    Full Text Available Obesity and diabetes are increasing at an alarming rate worldwide, but the strategies for the prevention and treatment of these disorders remain inadequate. Brown adipose tissue (BAT is important for cold protection by producing heat using lipids and glucose as metabolic fuels. This thermogenic action causes increased energy expenditure and significant lipid/glucose disposal. In addition, BAT in white adipose tissue (WAT or beige cells have been found and they also exhibit the thermogenic action similar to BAT. These data provide evidence indicating BAT/beige cells as a potential target for combating obesity and diabetes. Recent discoveries of active BAT and beige cells in adult humans have further highlighted this potential. Growing studies have also shown the importance of central nervous system in the control of BAT thermogenesis and WAT browning using animal models. This review is focused on central neural thermoregulation, particularly addressing our current understanding of the importance of hypothalamic neural signaling in the regulation of BAT/beige thermogenesis and energy homeostasis.

  12. Effects of a high-fat, low- versus high-glycemic index diet: retardation of insulin resistance involves adipose tissue modulation.

    Science.gov (United States)

    van Schothorst, Evert M; Bunschoten, Annelies; Schrauwen, Patrick; Mensink, Ronald P; Keijer, Jaap

    2009-04-01

    Beneficial effects of low glycemic index (GI) diets in rodents have been studied using healthy low-fat diets, while the effects might be different on high-fat diets inducing progression of insulin resistance. We fed C57BL/6J male mice high-fat low/high-GI (LGI/HGI) diets for 13 wk. Glucose and insulin tolerance and serum substrates, including adipokines, were measured longitudinally. The LGI group showed a significantly higher glucose tolerance from wk 2 onwards, which was supported by lower serum insulin and free fatty acids levels at 8 wk, and a tendency for lower leptin levels, while resistin levels remained similar. At 11 wk, when differences in serum resistin started to increase, differences in serum insulin were diminished. Although food intake was similar throughout the study, body weights and epididymal adipose tissue mass became significantly lower in the LGI group at necropsy. Several serum substrates and adipose tissue leptin mRNA levels, as analyzed by Q-PCR, were, again, significantly lower, whereas adiponectin mRNA levels were higher. Taken together, an LGI high-fat diet maintains higher glucose tolerance and insulin sensitivity via adipose tissue modulation solely because of a difference in the type of carbohydrate, supporting a nutritional approach in the fight against insulin resistance.

  13. Effects of exercise training on subcutaneous and visceral adipose tissue in normal- and high-fat diet-fed rats

    OpenAIRE

    Gollisch, Katja S. C.; Brandauer, Josef; Jessen, Niels; Toyoda, Taro; Nayer, Ali; Hirshman, Michael F.; Goodyear, Laurie J.

    2009-01-01

    Regular physical activity improves glucose tolerance and decreases adiposity. Our aim was to investigate the effects of exercise training on subcutaneous (inguinal) and visceral (parametrial) adipose tissue in rats that were fed a chow diet (13% fat) or made insulin resistant by a high-fat diet (60% fat). Sprague-Dawley rats performed 4 wk of voluntary wheel running or were kept as sedentary controls. The training groups fed chow and the high-fat diet achieved similar running distances (8.8 ±...

  14. Fungal endophytes in aboveground tissues of desert plants: infrequent in culture, but highly diverse and distinctive symbionts.

    Science.gov (United States)

    Massimo, Nicholas C; Nandi Devan, M M; Arendt, Kayla R; Wilch, Margaret H; Riddle, Jakob M; Furr, Susan H; Steen, Cole; U'Ren, Jana M; Sandberg, Dustin C; Arnold, A Elizabeth

    2015-07-01

    In hot deserts, plants cope with aridity, high temperatures, and nutrient-poor soils with morphological and biochemical adaptations that encompass intimate microbial symbioses. Whereas the root microbiomes of arid-land plants have received increasing attention, factors influencing assemblages of symbionts in aboveground tissues have not been evaluated for many woody plants that flourish in desert environments. We evaluated the diversity, host affiliations, and distributions of endophytic fungi associated with photosynthetic tissues of desert trees and shrubs, focusing on nonsucculent woody plants in the species-rich Sonoran Desert. To inform our strength of inference, we evaluated the effects of two different nutrient media, incubation temperatures, and collection seasons on the apparent structure of endophyte assemblages. Analysis of >22,000 tissue segments revealed that endophytes were isolated four times more frequently from photosynthetic stems than leaves. Isolation frequency was lower than expected given the latitude of the study region and varied among species a function of sampling site and abiotic factors. However, endophytes were very species-rich and phylogenetically diverse, consistent with less arid sites of a similar latitudinal position. Community composition differed among host species, but not as a function of tissue type, sampling site, sampling month, or exposure. Estimates of abundance, diversity, and composition were not influenced by isolation medium or incubation temperature. Phylogenetic analyses of the most commonly isolated genus (Preussia) revealed multiple evolutionary origins of desert-plant endophytism and little phylogenetic structure with regard to seasonality, tissue preference, or optimal temperatures and nutrients for growth in vitro. Together, these results provide insight into endophytic symbioses in desert-plant communities and can be used to optimize strategies for capturing endophyte biodiversity at regional scales.

  15. Deletion of serum amyloid A3 improves high fat high sucrose diet-induced adipose tissue inflammation and hyperlipidemia in female mice.

    Directory of Open Access Journals (Sweden)

    Laura J den Hartigh

    Full Text Available Serum amyloid A (SAA increases in response to acute inflammatory stimuli and is modestly and chronically elevated in obesity. SAA3, an inducible form of SAA, is highly expressed in adipose tissue in obese mice where it promotes monocyte chemotaxis, providing a mechanism for the macrophage accumulation that occurs with adipose tissue expansion in obesity. Humans do not express functional SAA3 protein, but instead express SAA1 and SAA2 in hepatic as well as extrahepatic tissues, making it difficult to distinguish between liver and adipose tissue-specific SAA effects. SAA3 does not circulate in plasma, but may exert local effects that impact systemic inflammation. We tested the hypothesis that SAA3 contributes to chronic systemic inflammation and adipose tissue macrophage accumulation in obesity using mice deficient for Saa3 (Saa3(-/-. Mice were rendered obese by feeding a pro-inflammatory high fat, high sucrose diet with added cholesterol (HFHSC. Both male and female Saa3(-/- mice gained less weight on the HFHSC diet compared to Saa3(+/+ littermate controls, with no differences in body composition or resting metabolism. Female Saa3(-/- mice, but not males, had reduced HFHSC diet-induced adipose tissue inflammation and macrophage content. Both male and female Saa3(-/- mice had reduced liver Saa1 and Saa2 expression in association with reduced plasma SAA. Additionally, female Saa3(-/- mice, but not males, showed improved plasma cholesterol, triglycerides, and lipoprotein profiles, with no changes in glucose metabolism. Taken together, these results suggest that the absence of Saa3 attenuates liver-specific SAA (i.e., SAA1/2 secretion into plasma and blunts weight gain induced by an obesogenic diet. Furthermore, adipose tissue-specific inflammation and macrophage accumulation are attenuated in female Saa3(-/- mice, suggesting a novel sexually dimorphic role for this protein. These results also suggest that Saa3 influences liver-specific SAA1

  16. [Expression of high mobility group box-1 in the lung tissue and serum of patients with pulmonary tuberculosis].

    Science.gov (United States)

    Yang, Xiao-min; Yang, Hua

    2013-07-01

    To explore the expression of high mobility group box-1 (HMGB1) in the lung tissue and serum of patients with pulmonary tuberculosis and to explore its relationship with tumor necrosis factor (TNF)-α and interleukin(IL)-1β. Sixty samples of lung tissues were obtained from patients with pulmonary tuberculosis who had underwent pneumonectomy in Department of Chest Surgery, First Affiliated Hospital of Zunyi Medical College from June 2010 to December 2011. At the same period, 40 normal lung samples were also obtained from patients with pulmonary contusion and lung cancer by surgical resections as the control group. The mRNA expressions of HMGB1 was detected by reverse transcription-polymerase chain reaction (RT-PCR), and the protein level of HMGB1 was measured by immunohistochemical staining of tissue microarrays in lung tissue. Blood samples were taken from 89 patients with active pulmonary tuberculosis (pulmonary tuberculosis group), including hematogenous disseminated pulmonary tuberculosis (type II) in 35 cases and secondary pulmonary tuberculosis (type III) in 54 cases, and 50 healthy volunteers (control group). Furthermore, the 54 patients with secondary pulmonary tuberculosis were divided into different subgroups according to cavity formation and the lung fields involved: patients without lung cavity (35 cases) vs those with lung cavity (19 cases), patients with involvement of pulmonary tuberculosis (69 ± 29) was significantly higher than that in normal lung tissue (22 ± 12) (t = 2.389, P pulmonary tuberculosis (786 ± 86) was significantly higher than that in normal lung tissue (202 ± 60) (t = 3.872, P pulmonary tuberculosis group were (5.0 ± 3.2) µg/L, (118 ± 77) ng/L and (33 ± 20) ng/L, respectively, which were significantly higher than those in the control group [(1.7 ± 1.0) µg/L, (40 ± 11) ng/L and (18 ± 12) ng/L, respectively], the respective t values being -0.928, 4.268 and 11.064, all P pulmonary tuberculosis, the serum concentration of HMGB

  17. Higher visceral adiposity is associated with an enhanced early thermogenic response to carbohydrate-rich food.

    Science.gov (United States)

    Gepner, Yftach; Bril, Nitzan; Shelef, Ilan; Schwarzfuchs, Dan; Serfaty, Dana; Rein, Michal; Cohen, Noa; Shemesh, Elad; Tangi-Rosental, Osnat; Sarusi, Benjamin; Goshen, Eyal; Kenigsbuch, Shira; Chassidim, Yoash; Golan, Rachel; Witkow, Shula; Henkin, Yaakov; Stampfer, Meir J; Rudich, Assaf; Shai, Iris

    2016-04-01

    Studies examining the dynamics of the thermic effect of feeding (TEF) of specific food items and the relationship of TEF to visceral adiposity are limited. We measured resting energy expenditure (REE) and early-TEF (40-min postprandial, e-TEF) after 8-h fast by indirect calorimetry in 40 obese men, and imaged abdominal fat tissues by magnetic resonance imaging. Each participant was examined on two occasions, 3-weeks apart. At each examination we measured fasting REE and then postprandial REE following the isocaloric [∼380 kcal] consumption of either 56 gr walnuts [(8% carbohydrates; 84% fat, of which 72% polyunsaturated fat)], or 5-slices (150gr) of whole-grain bread (48% carbohydrates; 32% fat). e-TEF was calculated as the area under the curve between the fasting and postprandial tests. Participants had a mean age of 45 ± 8 years, body-mass-index (BMI) = 31.1 ± 3.8 kg/m(2), total abdominal fat area = 901.4 ± 240 cm(2), visceral fat area (VAT) = 260 ± 102.9 cm(2), fasting REE = 1854 ± 205 kcal, REE/kg = 19.39 ± 1.73 kcal/kg, and respiratory quotient (RQ, CO2 eliminated/O2 consumed) = 0.82 ± 0.04. Individuals who exhibited increased e-TEF (top ΔAUC median) to bread had higher VAT (299 cm(2) vs. 223 cm(2); p = 0.024) and higher BMI (32.4 kg/m(2) vs. 30.0 kg/m(2); p = 0.013), compared to their peers with the lower e-TEF response (ΔAUC below median). As expected, postprandial e-TEF was higher after whole-grain bread consumption [ΔAUC = +14 kcal/40min] compared to walnuts [ΔAUC = -2 kcal/40 min; p food, likely reflecting digestion, early absorption and/or sympathetic tone (rather than metabolic utilization (oxidation)), associates with visceral adiposity. Future studies are required to determine if this association represents an added causality between early carbohydrate processing and visceral fat accumulation. Copyright © 2015 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All

  18. Predictors of local recurrence in high-grade soft tissue sarcomas: hydrogen peroxide as a local adjuvant.

    Science.gov (United States)

    Wooldridge, Adam N; Kolovich, Gregory P; Crist, Martha K; Mayerson, Joel L; Scharschmidt, Thomas J

    2013-02-01

    Soft tissue sarcomas have a mortality rate of 40% to 60%, with local recurrence being a poor prognostic factor for overall survival. Three-percent nondiluted hydrogen peroxide is hypothesized to be an effective local adjuvant. The purpose of this study was to identify risk factors for local recurrence in high-grade soft tissue sarcomas and to determine whether using hydrogen peroxide as a local adjuvant reduced the risks of local recurrence and surgical-site infection. Retrospective data were collected for 106 patients surgically treated for high-grade soft tissue sarcomas between 2002 and 2010. The primary endpoint was local recurrence. Eighteen (16.98%) cases of local recurrence occurred. Predictors of local recurrence were margin status, estimated blood loss, and histology (ie, malignant peripheral nerve sheath tumor), with hazard ratios of 4.44 (95% confidence interval [CI], 1.32-14.95), 1.19 (95% CI, 1.06-1.35), and 9.21 (95% CI, 2.11-40.16), respectively. Hydrogen peroxide yielded a statistically insignificant improvement in local recurrence, with a hazard ratio of 0.81 (95% CI, 0.27-2.48) and a reduced risk of surgical site infection, with a hazard ratio of 0.52 (95% CI, 0.15-1.81). Margin status, increased blood loss, and histologic subtype are associated with increased local recurrence risk. Using hydrogen peroxide improved local control and infection rates, but the difference was not statistically significant. Copyright 2013, SLACK Incorporated.

  19. A simple and efficient method for isolation of DNA in high mucilaginous plant tissues.

    Science.gov (United States)

    Echevarría-Machado, Ileana; Sánchez-Cach, Lucila A; Hernández-Zepeda, Cecilia; Rivera-Madrid, Renata; Moreno-Valenzuela, Oscar A

    2005-10-01

    A protocol is described for rapid DNA isolation from Malvaceae plant species and different tissues of Bixaceae that contain large amounts of polysaccharides, polyphenols, and pigments that interfere with DNA extractions. The method is a modification of Dellaporta et al. The current protocol is simple, and no phenol-chloroform extraction, ethanol, or isopropranol precipitation is required. The method is based in the incubation of soluble DNA with silica, mix in batch during the extraction. The procedure can be completed in 2 h and many samples can be processed at the same time. DNA of excellent quality was recovered and used for polymerase chain reaction (PCR) amplification, restriction enzyme digestion, and Southern blot analysis. The method was used with healthy Bixa orellana and virus-infected Malvaceae plants.

  20. Mammalian tissues defective in nonsense-mediated mRNA decay display highly aberrant splicing patterns

    DEFF Research Database (Denmark)

    Weischenfeldt, Joachim Lütken; Waage, Johannes Eichler; Tian, Geng

    2012-01-01

    a bioinformatic pipeline that maps RNA-seq data to a combinatorial exon database, predicts NMD-susceptibility for mRNA isoforms and calculates the distribution of major splice isoform classes. We present a catalog of NMD-regulated alternative splicing events, showing that isoforms of 30% of all expressed genes......ABSTRACT: BACKGROUND: Nonsense-mediated mRNA decay (NMD) affects the outcome of alternative splicing by degrading mRNA isoforms with premature termination codons. Splicing regulators constitute important NMD targets; however, the extent to which loss of NMD causes extensive deregulation...... of alternative splicing has not previously been assayed in a global, unbiased manner. Here, we combine mouse genetics and RNA-seq to provide the first in vivo analysis of the global impact of NMD on splicing patterns in two primary mouse tissues ablated for the NMD factor UPF2. RESULTS: We developed...

  1. Short Communication An efficient method for simultaneous extraction of high-quality RNA and DNA from various plant tissues.

    Science.gov (United States)

    Oliveira, R R; Viana, A J C; Reátegui, A C E; Vincentz, M G A

    2015-12-29

    Determination of gene expression is an important tool to study biological processes and relies on the quality of the extracted RNA. Changes in gene expression profiles may be directly related to mutations in regulatory DNA sequences or alterations in DNA cytosine methylation, which is an epigenetic mark. Correlation of gene expression with DNA sequence or epigenetic mark polymorphism is often desirable; for this, a robust protocol to isolate high-quality RNA and DNA simultaneously from the same sample is required. Although commercial kits and protocols are available, they are mainly optimized for animal tissues and, in general, restricted to RNA or DNA extraction, not both. In the present study, we describe an efficient and accessible method to extract both RNA and DNA simultaneously from the same sample of various plant tissues, using small amounts of starting material. The protocol was efficient in the extraction of high-quality nucleic acids from several Arabidopsis thaliana tissues (e.g., leaf, inflorescence stem, flower, fruit, cotyledon, seedlings, root, and embryo) and from other tissues of non-model plants, such as Avicennia schaueriana (Acanthaceae), Theobroma cacao (Malvaceae), Paspalum notatum (Poaceae), and Sorghum bicolor (Poaceae). The obtained nucleic acids were used as templates for downstream analyses, such as mRNA sequencing, quantitative real time-polymerase chain reaction, bisulfite treatment, and others; the results were comparable to those obtained with commercial kits. We believe that this protocol could be applied to a broad range of plant species, help avoid technical and sampling biases, and facilitate several RNA- and DNA-dependent analyses.

  2. Increased COX-2 expression in epithelial and stromal cells of high mammographic density tissues and in a xenograft model of mammographic density.

    Science.gov (United States)

    Chew, G L; Huo, C W; Huang, D; Hill, P; Cawson, J; Frazer, H; Hopper, J L; Haviv, I; Henderson, M A; Britt, K; Thompson, E W

    2015-08-01

    Mammographic density (MD) adjusted for age and body mass index is one of the strongest known risk factors for breast cancer. Given the high attributable risk of MD for breast cancer, chemoprevention with a safe and available agent that reduces MD and breast cancer risk would be beneficial. Cox-2 has been implicated in MD-related breast cancer risk, and was increased in stromal cells in high MD tissues in one study. Our study assessed differential Cox-2 expression in epithelial and stromal cells in paired samples of high and low MD human breast tissue, and in a validated xenograft biochamber model of MD. We also examined the effects of endocrine treatment upon Cox-2 expression in high and low MD tissues in the MD xenograft model. Paired high and low MD human breast tissue samples were immunostained for Cox-2, then assessed for differential expression and staining intensity in epithelial and stromal cells. High and low MD human breast tissues were separately maintained in biochambers in mice treated with Tamoxifen, oestrogen or placebo implants, then assessed for percentage Cox-2 staining in epithelial and stromal cells. Percentage Cox-2 staining was greater for both epithelial (p = 0.01) and stromal cells (p tissues. In high MD biochamber tissues, percentage Cox-2 staining was greater in stromal cells of oestrogen-treated versus placebo-treated tissues (p = 0.05).

  3. Effect of four weeks high intensity interval training versus aerobic exercise on metallothionein levels of myocardial tissue in rats

    Directory of Open Access Journals (Sweden)

    M. Shabani

    2016-08-01

    Full Text Available Background: Oxidative species produced during exercise can cause damages to some tissues such as kidney, liver and skeletal muscle. It seems that metallothionein plays a protective role against these actions. Objective: To assess the effect of four weeks of high intensity interval training versus aerobic exercise on metallothionein levels of myocardial tissue in healthy male rats. Methods: This experimental study was done on 36 male Wistar rats (2 months of age with an average weight of 180±20 in 2015.The rats were randomly divided into three groups: control (n=12, high intensity interval training (n=12, and aerobic training (n=12 groups. The experimental groups exercised according to the training program 5 days a week for 4 weeks. During this period, the control group did not have any training programs. One-way ANOVA was used to analyze the data. Findings: The results indicate no significant difference among the mean metallothionein in control, aerobic exercise and high intensity interval training groups. However, the amount of myocardial metallothionein increased as a result of high intensity interval training and decreased during aerobic exercise. Conclusion: It seems that the duration and intensity of exercise need to be adjusted in order to obtain better results.

  4. High-resolution and high sensitivity mesoscopic fluorescence tomography based on de-scanning EMCCD: System design and thick tissue imaging applications

    Science.gov (United States)

    Ozturk, Mehmet Saadeddin

    Optical microscopy has been one of the essential tools for biological studies for decades, however, its application areas was limited to superficial investigation due to strong scattering in live tissues. Even though advanced techniques such as confocal or multiphoton methods have been recently developed to penetrate beyond a few hundreds of microns deep in tissues, they still cannot perform in the mesoscopic regime (millimeter scale) without using destructive sample preparation protocols such as clearing techniques. They provide rich cellular information; however, they cannot be readily employed to investigate the biological processes at larger scales. Herein, we will present our effort to establish a novel imaging approach that can quantify molecular expression in intact tissues, well beyond the current microscopy depth limits. Mesoscopic Fluorescence Molecular Tomography (MFMT) is an emerging imaging modality that offers unique potential for the non-invasive molecular assessment of thick in-vitro and in-vivo live tissues. This novel imaging modality is based on an optical inverse problem that allows for retrieval of the quantitative spatial distribution of fluorescent tagged bio-markers at millimeter depth. MFMT is well-suited for in-vivo subsurface tissue imaging and thick bio-printed specimens due to its high sensitivity and fast acquisition times, as well as relatively large fields of view. Herein, we will first demonstrate the potential of this technique using our first generation MFMT system applied to multiplexed reporter gene imaging (in-vitro) and determination of Photodynamic Therapy (PDT) agent bio-distribution in a mouse model (in-vivo). Second, we will present the design rationale, in silico benchmarking, and experimental validation of a second generation MFMT (2GMFMT) system. We will demonstrate the gain in resolution and sensitivity achieved due to the de-scanned dense detector configuration implemented. The potential of this novel platform will be

  5. Glycemic index differences of high-fat diets modulate primarily lipid metabolism in murine adipose tissue

    NARCIS (Netherlands)

    Schothorst, van E.M.; Bunschoten, J.E.; Verlinde, E.; Schrauwen, P.; Keijer, J.

    2011-01-01

    A low vs. high glycemic index of a high-fat (HF) diet (LGI and HGI, respectively) significantly retarded adverse health effects in adult male C57BL/6J mice, as shown recently (Van Schothorst EM, Bunschoten A, Schrauwen P, Mensink RP, Keijer J. FASEB J 23: 1092–1101, 2009). The LGI diet enhanced

  6. High Quality RNA Isolation from Leaf, Shell, Root Tissues and Callus of Hazelnut (Corylus avellana L.

    Directory of Open Access Journals (Sweden)

    Hossein Khosravi

    2017-12-01

    Full Text Available Extraction of high quality RNA is a critical step in molecular genetics studies. Hazelnut is one of the most important nuts plants in the world. The presence of the taxol and other taxanes in hazelnut plant necessitates explaining their biosynthesis pathway and identifying the candidate genes. Therefore, an easy and practical method is necessary for RNA extraction from hazelnuts. Hazelnut has high levels of phenolic compounds. High amounts of polyphenolic and polysaccharide compounds in plants could be causing problems in RNA extraction procedures.  To avoid these problems, a simple and efficient method can be used based on cetyltrimethylammonium bromide (CTAB extraction buffer and lithium chloride for extraction of high quality RNA from different parts of hazelnut plant. Using this method, a high-quality RNA sample (light absorbed in the A260/A280 was 2.04

  7. Effect of continuous high intensity focused ultrasound in a squamous cell carcinoma tumor model compared to muscle tissue evaluated by MRI, histology, and gene expression.

    Science.gov (United States)

    Hundt, Walter; Yuh, Esther L; Steinbach, Silke; Bednarski, Mark D

    2009-04-01

    The purpose of this study was to investigate the effect of the continuous mode of high intensity focused ultrasound (HIFU) in a mouse head and neck cancer model (SCCVII) compared to muscle tissue. HIFU was applied to SCCVII tumors and to muscle tissue in C3H/Km mice using a dual ultrasound system (imaging 6 MHz/therapeutic 1 MHz). A continuous HIFU mode (total time 20 sec, intensity 6730.6 W/cm(2)) was applied. Three hours after HIFU treatment pre- and post-contrast T1-wt, T2-wt images, and a diffusion-wt STEAM sequence were obtained. After MR imaging, the animals were euthenized and the treated tumor and muscle tissue was taken out for histology and functional genomic analysis. T2 images showed increased signal intensity, post-contrast T1 showed a decreased contrast uptake in the central parts in the tumor tissue as well as in the muscle tissue. In addition a significant higher diffusion coefficient was found in both tissue types. Histological evaluation (H&E, Immunohistochemistry) of the tumors and the muscle tissue revealed areas of significant necrosis. In the tumor tissue 23 genes were up-regulated (> 2 fold change) and 4 genes were down-regulated (muscle tissue 29 genes were up-regulated and 17 genes down-regulated. Thirteen genes were up-regulated in both tissue types, 8 genes only in the SCCVII tissue, and 11 genes only in the muscle tissue. The use of HIFU treatment on tumor and muscle tissue results in dramatic changes in gene expression. The expression of some genes are tissue specific, the expression of other genes are independent of the tissue type.

  8. High dose Erythropoietin increases Brain Tissue Oxygen Tension in Severe Vasospasm after Subarachnoid Hemorrhage

    Directory of Open Access Journals (Sweden)

    Helbok Raimund

    2012-06-01

    Full Text Available Abstract Background Vasospasm-related delayed cerebral ischemia (DCI significantly impacts on outcome after aneurysmal subarachnoid hemorrhage (SAH. Erythropoietin (EPO may reduce the severity of cerebral vasospasm and improve outcome, however, underlying mechanisms are incompletely understood. In this study, the authors aimed to investigate the effect of EPO on cerebral metabolism and brain tissue oxygen tension (PbtO2. Methods Seven consecutive poor grade SAH patients with multimodal neuromonitoring (MM received systemic EPO therapy (30.000 IU per day for 3 consecutive days for severe cerebral vasospasm. Cerebral perfusion pressure (CPP, mean arterial blood pressure (MAP, intracranial pressure (ICP, PbtO2 and brain metabolic changes were analyzed during the next 24 hours after each dose given. Statistical analysis was performed with a mixed effects model. Results A total of 22 interventions were analyzed. Median age was 47 years (32–68 and 86 % were female. Three patients (38 % developed DCI. MAP decreased 2 hours after intervention (P btO2 significantly increased over time (P  Conclusions EPO increases PbtO2 in poor grade SAH patients with severe cerebral vasospasm. The effect on outcome needs further investigation.

  9. High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency.

    Science.gov (United States)

    Ayuso, E; Mingozzi, F; Montane, J; Leon, X; Anguela, X M; Haurigot, V; Edmonson, S A; Africa, L; Zhou, S; High, K A; Bosch, F; Wright, J F

    2010-04-01

    The purity of adeno-associated virus (AAV) vector preparations has important implications for both safety and efficacy of clinical gene transfer. Early-stage screening of candidates for AAV-based therapeutics ideally requires a purification method that is flexible and also provides vectors comparable in purity and potency to the prospective investigational product manufactured for clinical studies. The use of cesium chloride (CsCl) gradient-based protocols provides the flexibility for purification of different serotypes; however, a commonly used first-generation CsCl-based protocol was found to result in AAV vectors containing large amounts of protein and DNA impurities and low transduction efficiency in vitro and in vivo. Here, we describe and characterize an optimized, second-generation CsCl protocol that incorporates differential precipitation of AAV particles by polyethylene glycol, resulting in higher yield and markedly higher vector purity that correlated with better transduction efficiency observed with several AAV serotypes in multiple tissues and species. Vectors purified by the optimized CsCl protocol were found to be comparable in purity and functional activity to those prepared by more scalable, but less flexible serotype-specific purification processes developed for manufacture of clinical vectors, and are therefore ideally suited for pre-clinical studies supporting translational research.

  10. Neoadjuvant Chemotherapy with Ifosfamide, Cisplatin, Adriamycin and Mitomycin (IMAP for High risk Adult Soft Tissue Sarcomas

    Directory of Open Access Journals (Sweden)

    Mohagheghi Mohammad Ali

    2009-05-01

    Full Text Available To define efficacy of pre-operative chemotherapy in down staging of advanced non-round cell soft tissue sarcomas. From Sep 2002 to Dec 2005, 70 patients were treated by Ifosfamid, MESNA, cisplatin, adriamycin, mitomycin and subsequent surgery. Postoperatively, patients received radiotherapy in cases of microscopically incomplete resection or local recurrence. The median age of the patients was 34 years and the median tumor size was 14 cm. According to AJCC classification 46 patients had stage 3 and 24 had stage 4 diseases. The most common subtypes were MFH and leiomyosarcoma. The most common sites of tumors were lower extremity and trunk. Toxicity grades three or higher consisted of nausea, Leucopenia and infection. About 50% of the patients received G-CSF. Response to chemotherapy was assessable in 63 patients; 9 patients achieved complete response and 16 showed partial response. Disease progressed in 8 and did not change in 37. The best response was seen with MFH, fibrosarcoma and synovial sarcoma. After chemotherapy seventy percent of patients underwent complete surgery. Disease relapsed in 41 patients and twenty two patients died of metastasis. Median survival of patients was 30 months. IMAP plus G-CSF is safe and effective as preoperative chemotherapy in some subtypes of sarcomas, although the metastasis problem has not been eliminated

  11. Anterior perirectal fat tissue thickness is a strong predictor of recurrence after high-intensity focused ultrasound for prostate cancer.

    Science.gov (United States)

    Sumitomo, Makoto; Asakuma, Junichi; Yoshii, Hidehiko; Sato, Akinori; Horiguchi, Akio; Ito, Keiichi; Nagakura, Kazuhiko; Asano, Tomohiko

    2010-09-01

    To evaluate if and why obesity affects the clinical outcome in patients undergoing high-intensity focused ultrasound (HIFU) treatment for prostate cancer (CaP). 115 patients who underwent HIFU treatment for localized CaP were categorized as obese, overweight or normal according to body mass index (BMI). The thickness of the anterior perirectal fat tissue (APFT) was measured by transrectal ultrasonography. Treatment was considered to have failed in the case of biochemical failure according to the Phoenix definition, positive follow-up biopsy or initiation of salvage therapy. Cox proportional hazards analyses were used to identify possible predictors for disease free survival (DFS), and an experimental fat tissue model was made to evaluate the ablation effect at the target tissue. According to the classification by the Western Pacific Regional Office of WHO, 43 patients were of normal weight, 24 were overweight and 48 were obese. The BMI groups did not differ in Gleason score, prostate-specific antigen level at diagnosis or clinical stage. There were, however, significant correlations between BMI and prostate-specific antigen nadir (P APFT thickness (P APFT (P APFT thickness, for which obesity could be a useful surrogate, might represent the causative factor for poor clinical outcome after transrectal HIFU treatment for CaP.

  12. Adipose tissue transcriptomics and epigenomics in low birthweight men and controls: role of high-fat overfeeding.

    Science.gov (United States)

    Gillberg, Linn; Perfilyev, Alexander; Brøns, Charlotte; Thomasen, Martin; Grunnet, Louise G; Volkov, Petr; Rosqvist, Fredrik; Iggman, David; Dahlman, Ingrid; Risérus, Ulf; Rönn, Tina; Nilsson, Emma; Vaag, Allan; Ling, Charlotte

    2016-04-01

    Individuals who had a low birthweight (LBW) are at an increased risk of insulin resistance and type 2 diabetes when exposed to high-fat overfeeding (HFO). We studied genome-wide mRNA expression and DNA methylation in subcutaneous adipose tissue (SAT) after 5 days of HFO and after a control diet in 40 young men, of whom 16 had LBW. mRNA expression was analysed using Affymetrix Human Gene 1.0 ST arrays and DNA methylation using Illumina 450K BeadChip arrays. We found differential DNA methylation at 53 sites in SAT from LBW vs normal birthweight (NBW) men (false discovery rate obesity-related traits in a replication cohort of 142 individuals. DNA methylation at 652 CpG sites (including in CDK5, IGFBP5 and SLC2A4) was altered in SAT after overfeeding in this and in another cohort. Young men who had a LBW exhibit epigenetic alterations in their adipose tissue that potentially influence insulin resistance and risk of type 2 diabetes. Short-term overfeeding influences gene transcription and, to some extent, DNA methylation in adipose tissue; there was no major difference in this response between LBW and control participants.

  13. Pharmacokinetics and Tissue Distribution Study of Praeruptorin D from Radix Peucedani in Rats by High-Performance Liquid Chromatography (HPLC

    Directory of Open Access Journals (Sweden)

    Xue Du

    2012-07-01

    Full Text Available Praeruptorin D (PD, a major pyranocoumarin isolated from Radix Peucedani, exhibited antitumor and anti-inflammatory activities. The aim of this study was to investigate the pharmacokinetics and tissue distribution of PD in rats following intravenous (i.v. administration. The levels of PD in plasma and tissues were measured by a simple and sensitive reversed-phase high-performance liquid chromatography (HPLC method. The biosamples were treated by liquid-liquid extraction (LLE with methyl tert-butyl ether (MTBE and osthole was used as the internal standard (IS. The chromatographic separation was accomplished on a reversed-phase C18 column using methanol-water (75:25, v/v as mobile phase at a flow rate of 0.8 mL/min and ultraviolet detection wave length was set at 323 nm. The results demonstrate that this method has excellent specificity, linearity, precision, accuracy and recovery. The pharmacokinetic study found that PD fitted well into a two-compartment model with a fast distribution phase and a relative slow elimination phase. Tissue distribution showed that the highest concentration was observed in the lung, followed by heart, liver and kidney. Furthermore, PD can also be detected in the brain, which indicated that PD could cross the blood-brain barrier after i.v. administration.

  14. The design and development of a high-throughput magneto-mechanostimulation device for cartilage tissue engineering.

    Science.gov (United States)

    Brady, Mariea A; Vaze, Reva; Amin, Harsh D; Overby, Darryl R; Ethier, C Ross

    2014-02-01

    To recapitulate the in vivo environment and create neo-organoids that replace lost or damaged tissue requires the engineering of devices, which provide appropriate biophysical cues. To date, bioreactors for cartilage tissue engineering have focused primarily on biomechanical stimulation. There is a significant need for improved devices for articular cartilage tissue engineering capable of simultaneously applying multiple biophysical (electrokinetic and mechanical) stimuli. We have developed a novel high-throughput magneto-mechanostimulation bioreactor, capable of applying static and time-varying magnetic fields, as well as multiple and independently adjustable mechanical loading regimens. The device consists of an array of 18 individual stations, each of which uses contactless magnetic actuation and has an integrated Hall Effect sensing system, enabling the real-time measurements of applied field, force, and construct thickness, and hence, the indirect measurement of construct mechanical properties. Validation tests showed precise measurements of thickness, within 14 μm of gold standard calliper measurements; further, applied force was measured to be within 0.04 N of desired force over a half hour dynamic loading, which was repeatable over a 3-week test period. Finally, construct material properties measured using the bioreactor were not significantly different (p=0.97) from those measured using a standard materials testing machine. We present a new method for articular cartilage-specific bioreactor design, integrating combinatorial magneto-mechanostimulation, which is very attractive from functional and cost viewpoints.

  15. The effect of high-intensity training on mitochondrial fat oxidation in skeletal muscle and subcutaneous adipose tissue

    DEFF Research Database (Denmark)

    Larsen, Steen; Danielsen, J H; Søndergård, Stine Dam

    2015-01-01

    High-intensity interval training (HIT) is known to increase mitochondrial content in a similar way as endurance training [60-90% of maximal oxygen uptake (VO2peak )]. Whether HIT increases the mitochondria's ability to oxidize lipids is currently debated. We investigated the effect of HIT...... of HIT (three times per week at 298 ± 21 W). HIT significantly increased VO2peak from 2.9 ± 0.2 to 3.1 ± 0.2 L/min. No differences were seen in maximal fat oxidation in either skeletal muscle or adipose tissue. Km (app) for octanoyl carnitine or palmitoyl carnitine were similar after training in skeletal...... muscle and adipose tissue. Maximal OXPHOS capacity with complex I- and II-linked substrates was increased after training in skeletal muscle but not in adipose tissue. In conclusion, 6 weeks of HIT increased VO2peak . Mitochondrial content and mitochondrial OXPHOS capacity were increased in skeletal...

  16. Endothelial dysfunction in high fructose containing diet fed rats: Increased nitric oxide and decreased endothelin-1 levels in liver tissue

    Directory of Open Access Journals (Sweden)

    Zeki Arı

    2010-09-01

    Full Text Available Objectives: Dietary high fructose consumption which is closely associated with endothelial dysfunction via insulin re-sistance has recently increased in developed countries. Insulin resistance has a promoter effect on many metabolic disorders such as syndrome X, polycystic ovary syndrome, Type 2 diabetes mellitus etc. Our aim in this study is to understand the impact of increased fructose intake on metabolisms of glucose, insulin and endothelial dysfunction by measuring nitric oxide (NO and endothelin-1 (ET-1 levels in hepatic tissue which is crucial in fructose metabolism.Materials and Methods: We designed an animal study to understand increased fructose intake on hepatic endothe-lium. Twenty adult male albino rats were divided into two groups; the study group (group 1, n=10 received isocaloric fructose enriched diet (fructose-fed rats, containing 18.3% protein, 60.3% fructose and 5.2% fat while the control group received purified regular chow (group 2, n=10 for 2 weeks. After feeding period, blood and hepatic tissue samples were collected and glucose, insulin, NO and ET-1 levels were analysed.Results: We found increased fasting glucose and insulin levels and impaired glucose tolerance in fructose fed rats. Higher NO and lower ET–1 levels were also detected in hepatic tissue samples of the group 1.Conclusion: Increased fructose consumption has deleterious effects on glucose tolerance, insulin resistance and may cause to endothelial dysfunction.

  17. Radio Frequency Ultrasound Time Series Signal Analysis to Evaluate High-intensity Focused Ultrasound Lesion Formation Status in Tissue.

    Science.gov (United States)

    Mobasheri, Saeedeh; Behnam, Hamid; Rangraz, Parisa; Tavakkoli, Jahan

    2016-01-01

    High-intensity focused ultrasound (HIFU) is a novel treatment modality used by scientists and clinicians in the recent decades. This modality has had a great and significant success as a noninvasive surgery technique applicable in tissue ablation therapy and cancer treatment. In this study, radio frequency (RF) ultrasound signals were acquired and registered in three stages of before, during, and after HIFU exposures. Different features of RF time series signals including the sum of amplitude spectrum in the four quarters of the frequency range, the slope, and intercept of the best-fit line to the entire power spectrum and the Shannon entropy were utilized to distinguish between the HIFU-induced thermal lesion and the normal tissue. We also examined the RF data, frame by frame to identify exposure effects on the formation and characteristics of a HIFU thermal lesion at different time steps throughout the treatment. The results obtained showed that the spectrum frequency quarters and the slope and intercept of the best fit line to the entire power spectrum both increased two times during the HIFU exposures. The Shannon entropy, however, decreased after the exposures. In conclusion, different characteristics of RF time series signal possess promising features that can be used to characterize ablated and nonablated tissues and to distinguish them from each other in a quasi-quantitative fashion.

  18. Microfluidic 3D bone tissue model for high-throughput evaluation of wound-healing and infection-preventing biomaterials.

    Science.gov (United States)

    Lee, Joung-Hyun; Gu, Yexin; Wang, Hongjun; Lee, Woo Y

    2012-02-01

    We report the use of a microfluidic 3D bone tissue model, as a high-throughput means of evaluating the efficacy of biomaterials aimed at accelerating orthopaedic implant-related wound-healing while preventing bacterial infection. As an example of such biomaterials, inkjet-printed micropatterns were prepared to contain antibiotic and biphasic calcium phosphate (BCP) nanoparticles dispersed in a poly(D,L-lactic-co-glycolic) acid matrix. The micropatterns were integrated with a microfluidic device consisting of eight culture chambers. The micropatterns immediately and completely killed Staphylococcus epidermidis upon inoculation, and enhanced the calcified extracellular matrix production of osteoblasts. Without antibiotic elution, bacteria rapidly proliferated to result in an acidic microenvironment which was detrimental to osteoblasts. These results were used to demonstrate the tissue model's potential in: (i) significantly reducing the number of biomaterial samples and culture experiments required to assess in vitro efficacy for wound-healing and infection prevention and (ii) in situ monitoring of dynamic interactions of biomaterials with bacteria as wells as with tissue cells simultaneously. Copyright © 2011 Elsevier Ltd. All rights reserved.

  19. The acute effects of the thermogenic supplement Meltdown on energy expenditure, fat oxidation, and hemodynamic responses in young, healthy males

    Directory of Open Access Journals (Sweden)

    Cooke Matt

    2008-12-01

    Full Text Available Abstract The purpose of this study was to evaluate the effects of a thermogenic supplement, Meltdown, on energy expenditure, fat oxidation, and hemodynamics before and after maximal treadmill exercise. In a double-blind, randomized, placebo-controlled, cross-over design, 12 male participants underwent two testing sessions after consuming either the Meltdown or placebo supplement. While in a fasted state, participants rested for one hour, orally ingested either Meltdown or placebo and rested for another hour, performed a maximal treadmill exercise test, and then rested for another hour. Throughout the testing protocol, resting energy expenditure (REE and respiratory exchange ratio (RER were assessed. In addition, heart rate (HR and blood pressure (BP were assessed before and after exercise. Meltdown increased REE significantly more than placebo at 45 min (1.44 ± 0.25 vs. 1.28 ± 0.23 kcal/min; p = 0.003, 60 min (1.49 ± 0.28 vs. 1.30 ± 0.22 kcal/min; p = 0.025, and 120 min (1.51 ± 0.26 vs. 1.33 ± 0.27 kcals/min; p = 0.014 post-ingestion. Meltdown significantly decreased RER at 30 min (0.84 ± 0.03 vs. 0.91 ± 0.04; p = 0.022 and 45 min post-ingestion (0.82 ± 0.04 vs. 0.89 ± 0.05; p = 0.042, and immediately post-exercise (0.83 ± 0.05 vs. 0.90 ± 0.07; p = 0.009. Furthermore, over the course of the evaluation period, area under the curve assessment demonstrated that REE was significantly increased with Meltdown compared to placebo (992.5 ± 133.1 vs. 895.1 ± 296.1 kcals; p = 0.043, while RER was significantly less than placebo (5.55 ± 0.61 vs. 5.89 ± 0.44; p = 0.002 following ingestion. HR and BP were not significantly affected prior to exercise with either supplement (p > 0.05 and the exercise-induced increases for HR and BP decreased into recovery and were not different between supplements (p > 0.05. These data suggest that Meltdown enhances REE and fat oxidation more than placebo for several hours after ingestion in fully rested and

  20. Browning of white adipose tissue uncouples glucose uptake from insulin signaling.

    Directory of Open Access Journals (Sweden)

    Karin Mössenböck

    Full Text Available Presence of thermogenically active adipose tissue in adult humans has been inversely associated with obesity and type 2 diabetes. While it had been shown that insulin is crucial for the development of classical brown fat, its role in development and function of inducible brown-in-white (brite adipose tissue is less clear. Here we show that insulin deficiency impaired differentiation of brite adipocytes. However, adrenergic stimulation almost fully induced the thermogenic program under these settings. Although brite differentiation of adipocytes as well as browning of white adipose tissue entailed substantially elevated glucose uptake by adipose tissue, the capacity of insulin to stimulate glucose uptake surprisingly was not higher in the brite state. Notably, in line with the insulin-independent stimulation of glucose uptake, our data revealed that brite recruitment results in induction of solute carrier family 2 (GLUT-1 expression in adipocytes and inguinal WAT. These results for the first time demonstrate that insulin signaling is neither essential for brite recruitment, nor is it improved in cells or tissues upon browning.

  1. Canine mesenchymal stem cells are effectively labeled with silica nanoparticles and unambiguously visualized in highly autofluorescent tissues

    Directory of Open Access Journals (Sweden)

    Han Sei-Myoung

    2012-08-01

    Full Text Available Abstract Background Development of a method for long-term labeling of cells is critical to elucidate transplanted cell fate and migration as well as the contribution to tissue regeneration. Silica nanoparticles have been recently developed and demonstrated to be biocompatible with a high labeling capacity. Thus, our study was designed to assess the suitability of silica nanoparticles for labeling canine mesenchymal stem cells (MSCs and the fluorescence afficiency in highly autofluorescent tissue. Results We examined the effect of silica nanoparticle labeling on stem cell morphology, viability and differentiation as compared with those of unlabeled control cells. After 4 h of incubation with silica nanoparticles, they were internalized by canine MSCs without a change in the morphology of cells compared with that of control cells. The viability and proliferation of MSCs labeled with silica nanoparticles were evaluated by a WST-1 assay and trypan blue exclusion. No effects on cell viability were observed, and the proliferation of canine MSCs was not inhibited during culture with silica nanoparticles. Furthermore, adipogenic and osteogenic differentiation of silica nanoparticle-labeled canine MSCs was at a similar level compared with that of unlabeled cells, indicating that silica nanoparticle labeling did not alter the differentiation capacity of canine MSCs. Silica nanoparticle-labeled canine MSCs were injected into the kidneys of BALB/c mice after celiotomy, and then the mice were sacrificed after 2 or 3 weeks. The localization of injected MSCs was closely examined in highly autofluorescent renal tissues. Histologically, canine MSCs were uniformly and completely labeled with silica nanoparticles, and were unambiguously imaged in histological sections. Conclusions The results of the current study showed that silica nanoparticles are useful as an effective labeling marker for MSCs, which can elucidate the distribution and fate of transplanted

  2. Reply to the comment on ;First records of syn-diagenetic non-tectonic folding in Quaternary thermogene travertines caused by hydrothermal incremental veining; by Billi et alii

    Science.gov (United States)

    Billi, Andrea; Berardi, Gabriele; Gratier, Jean-Pierre; Rossetti, Federico; Vignaroli, Gianluca; Baykara, Mehmet Oruç; Bernasconi, Stefano M.; Kele, Sándor; Soligo, Michele; De Filippis, Luigi; Shen, Chuan-Chou

    2017-11-01

    In our previous paper (Billi et al., 2017), using field geological observations, U-Th dating, and stable isotope analyses, we studied two deposits of Pleistocene thermogene travertines from Tuscany in central Italy. We concluded our study (1) warning that the common stratigraphic concept of travertine being a sedimentary succession with age younging from bottom to top is not always correct, (2) demonstrating that CaCO3 mineralization and veins can develop within the travertines after their formation with this syn-diagenetic process being able to modify the continuous bottom-up age evolution, and (3) showing that this post-depositional mineralization-veining process can not only modify the temporal succession but also deform and change the initial depositional travertine structure and its petrophysical properties. These conclusions could potentially make the interpretation of a travertine series more difficult than commonly thought. Alcicek et alii questioned our conclusions claiming that the travertine structures that we observed in Tuscany and interpreted as post-depositional features should have been interpreted, in analogy to similar structures from travertines elsewhere, as primary structures. Although we recognize, as already thoroughly stated in Billi et al. (2017), that the travertine depositional/post-depositional processes generally require further studies, we reaffirm the validity of our original interpretation at least for the structures analyzed in our previous paper. We, therefore, counter all criticisms by Alcicek et alii and conclude by indicating the way forward to further explore the depositional and post-depositional processes of thermogene travertines.

  3. LIP volcanism and paleo-environmental crises - impact of magma emplacement sequence on thermogenic degassing rates from the Karoo sedimentary basin

    Science.gov (United States)

    Galerne, Christophe; Hasenclever, Jörg

    2017-04-01

    Volcanism in organic-rich sedimentary basins leading to thermogenic greenhouse gas generation has been documented as a strong forcing factor of past mass extinctions. However, quantitative studies fail to provide degassing rate estimates that would allow a direct comparison with anthropogenic warming. We are investigating different sill-emplacement sequences of a Large Igneous Province (LIP) plumbing system to identify their potential variable impact in terms of thermogenic degassing rates and cumulative amount of gas released at the basin top. We use a 2D finite element model that solves for hydrothermal fluid flow and thermal evolution around several cooling intrusions. Igneous sills are represented by horizontally dominated thermal anomalies that are sequentially placed within the sedimentary basin. We test different end-member scenarios of emplacement like bottom-to-top, top-to-bottom, and arbitrary emplacement order. Degassing pulses monitored during the simulations are recorded and compared for various end-member scenarios. The LIP emplaced in the Karoo Basin (South Africa, 183 Ma) is considered as a case study. We use basin lithostratigraphic properties (e.g. Total Organic Carbon content, sill to sediment proportion and structural data) to discuss results of our end-member models. This research potentially holds the key to demonstrate whether or not anthropogenic warming is in a comparable range to a documented paleo-environmental crisis and mass extinction triggered by degassing related volcanism.

  4. Curcuma longa extract associated with white pepper lessens high fat diet-induced inflammation in subcutaneous adipose tissue.

    Science.gov (United States)

    Neyrinck, Audrey M; Alligier, Maud; Memvanga, Patrick B; Névraumont, Elodie; Larondelle, Yvan; Préat, Véronique; Cani, Patrice D; Delzenne, Nathalie M

    2013-01-01

    Supra-nutritional doses of curcumin, derived from the spice Curcuma longa, have been proposed as a potential treatment of inflammation and metabolic disorders related to obesity. The aim of the present study was to test whether Curcuma longa extract rich in curcumin and associated with white pepper (Curcuma-P®), at doses compatible with human use, could modulate systemic inflammation in diet-induced obese mice. We questioned the potential relevance of changes in adiposity and gut microbiota in the effect of Curcuma-P® in obesity. Mice were fed either a control diet (CT), a high fat (HF) diet or a HF diet containing Curcuma longa extract (0.1 % of curcumin in the HF diet) associated with white pepper (0.01 %) for four weeks. Curcumin has been usually combined with white pepper, which contain piperine, in order to improve its bioavailability. This combination did not significantly modify body weight gain, glycemia, insulinemia, serum lipids and intestinal inflammatory markers. Tetrahydrocurcumin, but not curcumin accumulated in the subcutaneous adipose tissue. Importantly, the co-supplementation in curcuma extract and white pepper decreased HF-induced pro-inflammatory cytokines expression in the subcutaneous adipose tissue, an effect independent of adiposity, immune cells recruitment, angiogenesis, or modulation of gut bacteria controlling inflammation. These findings support that nutritional doses of Curcuma longa, associated with white pepper, is able to decrease inflammatory cytokines expression in the adipose tissue and this effect could be rather linked to a direct effect of bioactive metabolites reaching the adipose tissue, than from changes in the gut microbiota composition.

  5. Curcuma longa extract associated with white pepper lessens high fat diet-induced inflammation in subcutaneous adipose tissue.

    Directory of Open Access Journals (Sweden)

    Audrey M Neyrinck

    Full Text Available Supra-nutritional doses of curcumin, derived from the spice Curcuma longa, have been proposed as a potential treatment of inflammation and metabolic disorders related to obesity. The aim of the present study was to test whether Curcuma longa extract rich in curcumin and associated with white pepper (Curcuma-P®, at doses compatible with human use, could modulate systemic inflammation in diet-induced obese mice. We questioned the potential relevance of changes in adiposity and gut microbiota in the effect of Curcuma-P® in obesity.Mice were fed either a control diet (CT, a high fat (HF diet or a HF diet containing Curcuma longa extract (0.1 % of curcumin in the HF diet associated with white pepper (0.01 % for four weeks. Curcumin has been usually combined with white pepper, which contain piperine, in order to improve its bioavailability. This combination did not significantly modify body weight gain, glycemia, insulinemia, serum lipids and intestinal inflammatory markers. Tetrahydrocurcumin, but not curcumin accumulated in the subcutaneous adipose tissue. Importantly, the co-supplementation in curcuma extract and white pepper decreased HF-induced pro-inflammatory cytokines expression in the subcutaneous adipose tissue, an effect independent of adiposity, immune cells recruitment, angiogenesis, or modulation of gut bacteria controlling inflammation.These findings support that nutritional doses of Curcuma longa, associated with white pepper, is able to decrease inflammatory cytokines expression in the adipose tissue and this effect could be rather linked to a direct effect of bioactive metabolites reaching the adipose tissue, than from changes in the gut microbiota composition.

  6. Calorie Restriction with a High-Fat Diet Effectively Attenuated Inflammatory Response and Oxidative Stress-Related Markers in Obese Tissues of the High Diet Fed Rats

    Directory of Open Access Journals (Sweden)

    Seungae Park

    2012-01-01

    Full Text Available Obesity characterized by increased mass of adipose tissue leads to systemic inflammation. Calorie restriction (CR improves parameters associated with immune response and antioxidant defense. We hypothesized that CR with a high fat diet (HFCR regulates local and systemic inflammation and oxidative stress damage in a high fat diet induced obesity (HF group. We investigated effect of HFCR on inflammation and oxidative stress-related markers in liver and adipose tissues as well as adipokines in plasma. HFCR lowered liver triglyceride levels, total cholesterol levels, and the plasma leptin/adiponectin ratio to normal levels and improved glucose tolerance. HFCR also improved fatty liver and normalized adipocyte size and morphology. HFCR reduced lipid peroxidation and decreased the expression levels of inducible nitric oxide synthetase, cyclooxygenase-2, NF-E2-related factor, and heme oxygenase-1 in the liver. Moreover, HFCR suppressed the expression levels of C- reactive protein and manganese superoxide dismutase in the adipose tissue in the HF group. These results suggest that HFCR may have beneficial effects on inflammation and oxidative stress as well as lipid profiles in the HF diet induced obesity. Moreover, HFCR may be a good way to increase compliance in obese patients and to prevent obesity induced complications without changes in dietary pattern.

  7. Protein profile study of Pap smear and tissue of cervix by high performance liquid chromatography: laser induced fluorescence

    Science.gov (United States)

    Sujatha, N.; Rai, Lavanya; Kumar, Pratap; Krishnanand, B. R.; Mahato, K. K.; George, Sajan D.; Kartha, V. B.; C, Santhosh

    2007-02-01

    HPLC combined with laser induced fluorescence provides a very sensitive method for the separation and identification of the many proteins present in clinical samples. Protein profiles of clinical samples like Pap smear and tissue samples, from subjects with cervical cancer and normal volunteers, were recorded using HPLC-LIF. The protein profiles were analyzed by Principal Component Analysis (PCA). The profiles were characterized by parameters like scores of the factors, sum of squared residuals, and Mahalanobis Distance, derived from PCA. Parameters of each sample were compared with those of a standard set and Match/ No Match results were generated. Good discrimination between normal and malignant samples was achieved with high sensitivity and specificity.

  8. Biodegradable, Elastomeric, and Intrinsically Photoluminescent Poly(Silicon-Citrates) with high Photostability and Biocompatibility for Tissue Regeneration and Bioimaging.

    Science.gov (United States)

    Du, Yuzhang; Xue, Yumeng; Ma, Peter X; Chen, Xiaofeng; Lei, Bo

    2016-02-04

    Biodegradable polymer biomaterials with intrinsical photoluminescent properties have attracted much interest, due to their potential advantages for tissue regeneration and noninvasive bioimaging. However, few of current biodegradable polymers possess tunable intrinsically fluorescent properties, such as high photostability, fluorescent lifetime, and quantum field, and strong mechanical properties for meeting the requirements of biomedical applications. Here, by a facile one-step thermal polymerization, elastomeric poly(silicone-citrate) (PSC) hybrid polymers are developed with controlled biodegradability and mechanical properties, tunable inherent fluorescent emission (up to 600 nm), high photostability (beyond 180 min for UV and six months for natural light), fluorescent lifetime (near 10 ns) and quantum yield (16%-35%), high cellular biocompatibility, and minimal inflammatory response in vivo, which provide advantages over conventional fluorescent dyes, quantum dots, and current fluorescent polymers. The promising applications of PSC hybrids for cell and implants imaging in vitro and in vivo are successfully demonstrated. The development of elastomeric PSC polymer may provide a new strategy in synthesizing new inorganic-organic hybrid photo-luminescent materials for tissue regeneration and bioimaging applications. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Hypoxia-inducible factor 1α predicts recurrence in high-grade soft tissue sarcoma of extremities and trunk wall

    DEFF Research Database (Denmark)

    Nyström, Harriet; Jönsson, M; Werner-Hartman, L

    2017-01-01

    BACKGROUND AND AIM: Sarcomas are of mesenchymal origin and typically show abundant tumour stroma and presence of necrosis. In search for novel biomarkers for personalised therapy, we determined the prognostic impact of stromal markers, hypoxia and neovascularity in high-grade soft tissue leiomyos......BACKGROUND AND AIM: Sarcomas are of mesenchymal origin and typically show abundant tumour stroma and presence of necrosis. In search for novel biomarkers for personalised therapy, we determined the prognostic impact of stromal markers, hypoxia and neovascularity in high-grade soft tissue...... leiomyosarcoma and pleomorphic undifferentiated sarcoma. METHOD: We evaluated CD163, colony-stimulating factor (CSF)-1, CD16 and hypoxia-inducible factor 1 (HIF-1)α using immunohistochemical staining and assessed microvessel density using CD31 in 73 high-grade leiomyosarcomas and undifferentiated pleomorphic...... sarcomas of the extremities and the trunk wall. The results were correlated to metastasis-free and overall survival. RESULTS: Expression of HIF-1α was associated with the presence of necrosis and independently predicted shorter metastasis-free survival (HR 3.2, CI 1.4 to 7.0, p=0.004), whereas neither...

  10. Dose optimization of intra-operative high dose rate interstitial brachytherapy implants for soft tissue sarcoma

    Directory of Open Access Journals (Sweden)

    Jamema Swamidas

    2009-01-01

    Full Text Available Objective : A three dimensional (3D image-based dosimetric study to quantitatively compare geometric vs. dose-point optimization in combination with graphical optimization for interstitial brachytherapy of soft tissue sarcoma (STS. Materials and Methods : Fifteen consecutive STS patients, treated with intra-operative, interstitial Brachytherapy, were enrolled in this dosimetric study. Treatment plans were generated using dose points situated at the "central plane between the catheters", "between the catheters throughout the implanted volume", at "distances perpendicular to the implant axis" and "on the surface of the target volume" Geometrically optimized plans had dose points defined between the catheters, while dose-point optimized plans had dose points defined at a plane perpendicular to the implant axis and on the target surface. Each plan was graphically optimized and compared using dose volume indices. Results : Target coverage was suboptimal with coverage index (CI = 0.67 when dose points were defined at the central plane while it was superior when the dose points were defined at the target surface (CI=0.93. The coverage of graphically optimized plans (GrO was similar to non-GrO with dose points defined on surface or perpendicular to the implant axis. A similar pattern was noticed with conformity index (0.61 vs. 0.82. GrO were more conformal and less homogeneous compared to non-GrO. Sum index was superior for dose points defined on the surface of the target and relatively inferior for plans with dose points at other locations (1.35 vs. 1.27. Conclusions : Optimization with dose points defined away from the implant plane and on target results in superior target coverage with optimal values of other indices. GrO offer better target coverage for implants with non-uniform geometry and target volume.

  11. Antibody response to polyomavirus primary infection: high seroprevalence of Merkel cell polyomavirus and lymphoid tissue involvement.

    Science.gov (United States)

    Cason, Carolina; Monasta, Lorenzo; Zanotta, Nunzia; Campisciano, Giuseppina; Maestri, Iva; Tommasino, Massimo; Pawlita, Michael; Villani, Sonia; Comar, Manola; Delbue, Serena

    2018-01-12

    Human polyomaviruses (HPyVs) asymptomatically infect the human population establishing latency in the host, and their seroprevalence can reach 90% in healthy adults. Few studies have focused on the pediatric population, and there are no reports regarding the seroprevalence of all the newly isolated HPyVs among Italian children. Therefore, we investigated the frequency of serum antibodies against 12 PyVs in 182 immunocompetent children from Northeast Italy, by means of a multiplex antibody detection system. Additionally, secondary lymphoid tissues were collected to analyze the presence of HPyV DNA sequences using a specific real-time PCRs or PCRs. Almost 100% of subjects were seropositive for at least one PyV. Seropositivity ranged from 3% for antibodies against simian virus 40 (SV40) in children from 0 to 3 years, to 91% for antibodies against WU polyomavirus (WUPyV) and HPyV10 in children from 8 to 17 years. The mean number of PyV for which children were seropositive increased with the increasing of age: 4 standard deviations (SD) 1.8 in the 0-3-year group, 5 (SD 1.9) in the 4-7-year group, and 6 (SD 2.2) in the 8-17-year group. JC polyomavirus (JCPyV) DNA was detected in 1% of the adenoids, WUPyV in 12% of the tonsils, and 28% of the adenoids, and Merkel cell polyomavirus (MCPyV) was present in 6 and 2% of the tonsils and adenoids, respectively. Our study gives new insights on the serological evidence of exposure to PyVs during childhood, and on their possible respiratory route of transmission.

  12. High risk human papilloma viruses (HPVs) are present in benign prostate tissues before development of HPV associated prostate cancer.

    Science.gov (United States)

    Glenn, Wendy K; Ngan, Christopher C; Amos, Timothy G; Edwards, Richard J; Swift, Joshua; Lutze-Mann, Louise; Shang, Fei; Whitaker, Noel J; Lawson, James S

    2017-01-01

    Although high risk HPVs are associated with an increased risk of prostate cancer it is not known if they have a causal role. The purpose of this study is to investigate the potential role of human papilloma viruses (HPVs) in prostate cancer. The aims are (i) to investigate the presence and confirm the identity of high risk HPVs in benign prostate tissues prior to the development of HPV positive prostate cancer in the same patients, and (ii) to determine if HPVs are biologically active. We used polymerase chain reaction (PCR) to identify HPVs in specimens from 52 Australian men with benign prostate biopsies who 1 to 10 years later developed prostate cancer. Immunohistochemistry (IHC) was used to assess the expression of HPV E7 oncoproteins, cytokeratin and prostate specific antigen (PSA). We used RNASeq data from The Cancer Genome Atlas (TCGA) to identify possible HPV RNA sequences in prostate cancer. HPV screening using standard PCR was conducted on 28 of the 52 sets of benign and later prostate cancers. HPV L1 genes were identified in 13 (46%) benign and 8 (29%) of 28 later prostate cancers in the same patients. HPV E7 genes were identified in 23 (82%) benign and 19 (68%) of 28 subsequent prostate cancers in the same patients. The same HPV types were present in both the benign and subsequent prostate cancers in 9 sets of specimens. HPV type 16 was identified in 15% of benign and 3% of prostate cancers. HPV type 18 was identified in 26% of benign and 16% of prostate cancers. Small numbers of HPV types 45, 47, 76 and 115 were also identified. High confidence RNA-Seq evidence for high risk HPV types 16 and 18 was identified in 12 (2%) of the 502 TCGA prostate cancer transcriptomes. High risk HPV E7 oncoprotein was positively expressed in 23 (82%) of 28 benign prostate specimens but only in 8 (29%) of 28 of the later prostate cancer specimens. This difference is statistically significant (p = 0.001). Prostate specific antigen (PSA) was more highly expressed in 26

  13. Histological validation of high-resolution DTI in human post mortem tissue.

    Science.gov (United States)

    Seehaus, Arne; Roebroeck, Alard; Bastiani, Matteo; Fonseca, Lúcia; Bratzke, Hansjürgen; Lori, Nicolás; Vilanova, Anna; Goebel, Rainer; Galuske, Ralf

    2015-01-01

    Diffusion tensor imaging (DTI) is amongst the simplest mathematical models available for diffusion magnetic resonance imaging, yet still by far the most used one. Despite the success of DTI as an imaging tool for white matter fibers, its anatomical underpinnings on a microstructural basis remain unclear. In this study, we used 65 myelin-stained sections of human premotor cortex to validate modeled fiber orientations and oft used microstructure-sensitive scalar measures of DTI on the level of individual voxels. We performed this validation on high spatial resolution diffusion MRI acquisitions investigating both white and gray matter. We found a very good agreement between DTI and myelin orientations with the majority of voxels showing angular differences less than 10°. The agreement was strongest in white matter, particularly in unidirectional fiber pathways. In gray matter, the agreement was good in the deeper layers highlighting radial fiber directions even at lower fractional anisotropy (FA) compared to white matter. This result has potentially important implications for tractography algorithms applied to high resolution diffusion MRI data if the aim is to move across the gray/white matter boundary. We found strong relationships between myelin microstructure and DTI-based microstructure-sensitive measures. High FA values were linked to high myelin density and a sharply tuned histological orientation profile. Conversely, high values of mean diffusivity (MD) were linked to bimodal or diffuse orientation distributions and low myelin density. At high spatial resolution, DTI-based measures can be highly sensitive to white and gray matter microstructure despite being relatively unspecific to concrete microarchitectural aspects.

  14. Added thermogenic and satiety effects of a mixed nutrient vs a sugar-only beverage.

    Science.gov (United States)

    St-Onge, M-P; Rubiano, F; DeNino, W F; Jones, A; Greenfield, D; Ferguson, P W; Akrabawi, S; Heymsfield, S B

    2004-02-01

    To examine the effects of a sugar-only (SO) beverage vs one containing a mixed-nutrient (MN) composition on energy expenditure and feelings of hunger and satiety. A beverage containing a mixed macronutrient composition will lead to greater thermic effect of food and feelings of fullness than an isocaloric beverage containing only sugar. Adults were randomly assigned to receive a 2510 kJ (600 kcal) SO liquid formula followed by an isovolumic, isoenergetic, MN liquid formula with an energy distribution of 17% protein, 67% carbohydrates as sucrose and corn syrup solids, and 16% fat, or vice versa, in a crossover design. The carbohydrate source in the two beverages was identical: 1:1 ratio of sucrose and corn syrup solids (25 dextrose equivalents). The thermic response was calculated as the 7 h deviation from resting metabolic rate (RMR). Subjects provided hunger/satiety ratings and other related information by visual analog scales at regular intervals throughout the study period. In all, 20 subjects completed the protocol; one was removed from the thermic effect analysis due to discrepant RMRs. Following beverage ingestion, SO and MN liquid meals produced 7 h thermic effects of (X+/-s.e.m.) 274.1+/-27.6 kJ (65.5+/-6.6 kcal) and 372.0+/-33.9 kJ (88.9+/-8.1 kcal), respectively, resulting in a significant (P<0.01) difference between meals (Delta=97.9+/-35.1 kJ [23.4+/-8.4 kcal]). Analysis of satiety ratings using area under the curve analysis showed greater feelings of satiety (P<0.05) with MN compared to SO consumption. Also, subjects felt that they could eat less (P<0.05) after consumption of the MN vs SO beverage. In comparison to MN beverages, SO beverages are associated with a relatively high-energy retention without accompanying subjective hunger/fullness compensations, suggesting a basis for their role in long-term unintentional weight gain in healthy adults.

  15. High intravascular tissue factor expression in dogs with idiopathic immune-mediated haemolytic anaemia

    NARCIS (Netherlands)

    Piek, C.J.; Brinkhof, B.; Teske, E.; Rothuizen, J.; Dekker, A.; Penning, L.C.

    2011-01-01

    A high mortality occurs in dogs with idiopathic immune-mediated haemolytic anaemia (IMHA) during the first 2 weeks after the diagnosis. The aim of this study was to investigate the inflammatory response and coagulation abnormalities in dogs with IMHA in relation to the prognosis and to establish the

  16. High-frequency ultrasound for monitoring changes in liver tissue during preservation

    Science.gov (United States)

    Vlad, Roxana M.; Czarnota, Gregory J.; Giles, Anoja; Sherar, Michael D.; Hunt, John W.; Kolios, Michael C.

    2005-01-01

    Currently the only method to assess liver preservation injury is based on liver appearance and donor medical history. Previous work has shown that high-frequency ultrasound could detect ischemic cell death due to changes in cell morphology. In this study, we use high-frequency ultrasound integrated backscatter to assess liver damage in experimental models of liver ischemia. Ultimately, our goal is to predict organ suitability for transplantation using high-frequency imaging and spectral analysis techniques. To examine the effects of liver ischemia at different temperatures, livers from Wistar rats were surgically excised, immersed in phosphate buffer saline and stored at 4 and 20 °C for 24 h. To mimic organ preservation, livers were excised, flushed with University of Wisconsin (UW) solution and stored at 4 °C for 24 h. Preservation injury was simulated by either not flushing livers with UW solution or, before scanning, allowing livers to reach room temperature. Ultrasound images and corresponding radiofrequency data were collected over the ischemic period. No significant increase in integrated backscatter (~2.5 dBr) was measured for the livers prepared using standard preservation conditions. For all other ischemia models, the integrated backscatter increased by 4-9 dBr demonstrating kinetics dependent on storage conditions. The results provide a possible framework for using high-frequency imaging to non-invasively assess liver preservation injury.

  17. High-frequency ultrasound for monitoring changes in liver tissue during preservation

    Energy Technology Data Exchange (ETDEWEB)

    Vlad, Roxana M [Department of Medical Biophysics, University of Toronto, Toronto, Ontario (Canada); Czarnota, Gregory J [Department of Radiation Oncology, University of Toronto, Toronto, Ontario (Canada); Giles, Anoja [Department of Medical Biophysics, University of Toronto, Toronto, Ontario (Canada); Sherar, Michael D [Department of Medical Biophysics, University of Toronto, Toronto, Ontario (Canada); Hunt, John W [Department of Medical Biophysics, University of Toronto, Toronto, Ontario (Canada); Kolios, Michael C [Department of Medical Biophysics, University of Toronto, Toronto, Ontario (Canada)

    2005-01-21

    Currently the only method to assess liver preservation injury is based on liver appearance and donor medical history. Previous work has shown that high-frequency ultrasound could detect ischemic cell death due to changes in cell morphology. In this study, we use high-frequency ultrasound integrated backscatter to assess liver damage in experimental models of liver ischemia. Ultimately, our goal is to predict organ suitability for transplantation using high-frequency imaging and spectral analysis techniques. To examine the effects of liver ischemia at different temperatures, livers from Wistar rats were surgically excised, immersed in phosphate buffer saline and stored at 4 and 20 deg. C for 24 h. To mimic organ preservation, livers were excised, flushed with University of Wisconsin (UW) solution and stored at 4 deg. C for 24 h. Preservation injury was simulated by either not flushing livers with UW solution or, before scanning, allowing livers to reach room temperature. Ultrasound images and corresponding radiofrequency data were collected over the ischemic period. No significant increase in integrated backscatter ({approx}2.5 dBr) was measured for the livers prepared using standard preservation conditions. For all other ischemia models, the integrated backscatter increased by 4-9 dBr demonstrating kinetics dependent on storage conditions. The results provide a possible framework for using high-frequency imaging to non-invasively assess liver preservation injury.

  18. Development and validation of a high performance liquid chromatography method to determine linezolid concentrations in pig pulmonary tissue.

    Science.gov (United States)

    Guerrero, Laura; Martínez-Olondris, Pilar; Rigol, Montserrat; Esperatti, Mariano; Esquinas, Cristina; Luque, Néstor; Piñer, Raquel; Torres, Antoni; Soy, Dolors

    2010-03-01

    Linezolid is the first synthetic compound of a new group of antimicrobials, the oxazolidinones, which inhibit protein synthesis. It shows a broad spectrum of activity against Gram positive organisms. With respect to its pharmacokinetics, linezolid shows a relatively high volume of distribution and good penetration into inflammatory fluids, bone, fat and muscle. A reversed-phase isocratic high-performance liquid chromatographic method for linezolid analysis in piglet pulmonary tissue is described. Tissue samples and controls were prepared in 1 x TBE (1 M Tris, 0.9 M boric acid, 0.01 M EDTA). The mobile phase consisted of 20% ultrafiltered water and 80% of (A) 15 mM potassium monohydrogen phosphate buffer (pH = 5) with (B) acetonitrile (80%/20%; v/v). Samples were homogenized and precipitated with HClO(4) 3% (1/1, v/v). The injection volume was 100 microL. Ofloxacin was used as an internal standard. The assay was linear over a linezolid concentration range: 1.6-100 microg/mL. The method provided good validation data (n = 15): inaccuracy (3.6%), intra and inter-day variability (4.2% and 5.2%, respectively), recovery (91.8%), limit of detection (0.8 microg/mL) and quantitation (1.6 microg/mL) and acceptable stability within 24 h in the auto-sampler. The method offers a fast and simple approach to determine linezolid in pulmonary tissue which could be of use in pharmacokinetic studies.

  19. Effect of dietary krill oil supplementation on the endocannabinoidome of metabolically relevant tissues from high-fat-fed mice

    Directory of Open Access Journals (Sweden)

    Cohn Jeffrey S

    2011-07-01

    Full Text Available Abstract Background Omega-3 polyunsaturated fatty acids (ω-3-PUFA are known to ameliorate several metabolic risk factors for cardiovascular disease, and an association between elevated peripheral levels of endogenous ligands of cannabinoid receptors (endocannabinoids and the metabolic syndrome has been reported. We investigated the dose-dependent effects of dietary ω-3-PUFA supplementation, given as krill oil (KO, on metabolic parameters in high fat diet (HFD-fed mice and, in parallel, on the levels, in inguinal and epididymal adipose tissue (AT, liver, gastrocnemius muscle, kidneys and heart, of: 1 the endocannabinoids, anandamide and 2-arachidonoylglycerol (2-AG, 2 two anandamide congeners which activate PPARα but not cannabinoid receptors, N-oleoylethanolamine and N-palmitoylethanolamine, and 3 the direct biosynthetic precursors of these compounds. Methods Lipids were identified and quantified using liquid chromatography coupled to atmospheric pressure chemical ionization single quadrupole mass spectrometry (LC-APCI-MS or high resolution ion trap-time of flight mass spectrometry (LC-IT-ToF-MS. Results Eight-week HFD increased endocannabinoid levels in all tissues except the liver and epididymal AT, and KO reduced anandamide and/or 2-AG levels in all tissues but not in the liver, usually in a dose-dependent manner. Levels of endocannabinoid precursors were also generally down-regulated, indicating that KO affects levels of endocannabinoids in part by reducing the availability of their biosynthetic precursors. Usually smaller effects were found of KO on OEA and PEA levels. Conclusions Our data suggest that KO may promote therapeutic benefit by reducing endocannabinoid precursor availability and hence endocannabinoid biosynthesis.

  20. Effect of dietary krill oil supplementation on the endocannabinoidome of metabolically relevant tissues from high-fat-fed mice.

    Science.gov (United States)

    Piscitelli, Fabiana; Carta, Gianfranca; Bisogno, Tiziana; Murru, Elisabetta; Cordeddu, Lina; Berge, Kjetil; Tandy, Sally; Cohn, Jeffrey S; Griinari, Mikko; Banni, Sebastiano; Di Marzo, Vincenzo

    2011-07-13

    Omega-3 polyunsaturated fatty acids (ω-3-PUFA) are known to ameliorate several metabolic risk factors for cardiovascular disease, and an association between elevated peripheral levels of endogenous ligands of cannabinoid receptors (endocannabinoids) and the metabolic syndrome has been reported. We investigated the dose-dependent effects of dietary ω-3-PUFA supplementation, given as krill oil (KO), on metabolic parameters in high fat diet (HFD)-fed mice and, in parallel, on the levels, in inguinal and epididymal adipose tissue (AT), liver, gastrocnemius muscle, kidneys and heart, of: 1) the endocannabinoids, anandamide and 2-arachidonoylglycerol (2-AG), 2) two anandamide congeners which activate PPARα but not cannabinoid receptors, N-oleoylethanolamine and N-palmitoylethanolamine, and 3) the direct biosynthetic precursors of these compounds. Lipids were identified and quantified using liquid chromatography coupled to atmospheric pressure chemical ionization single quadrupole mass spectrometry (LC-APCI-MS) or high resolution ion trap-time of flight mass spectrometry (LC-IT-ToF-MS). Eight-week HFD increased endocannabinoid levels in all tissues except the liver and epididymal AT, and KO reduced anandamide and/or 2-AG levels in all tissues but not in the liver, usually in a dose-dependent manner. Levels of endocannabinoid precursors were also generally down-regulated, indicating that KO affects levels of endocannabinoids in part by reducing the availability of their biosynthetic precursors. Usually smaller effects were found of KO on OEA and PEA levels. Our data suggest that KO may promote therapeutic benefit by reducing endocannabinoid precursor availability and hence endocannabinoid biosynthesis.

  1. High-dose-rate intraoperative brachytherapy (IOHDR) using flab technique in the treatment of soft tissue sarcomas

    Energy Technology Data Exchange (ETDEWEB)

    Rachbauer, F.; Sununu, T.; Bach, C.; Nogler, M.; Krismer, M. [Dept. of Orthopedics, Univ. of Innsbruck (Austria); Sztankay, A.; Eichberger, P.; Schiestl, B.; Lukas, P. [Dept. of Radiotherapy-Radiooncology, Univ. of Innsbruck (Austria); Kreczy, A. [Inst. of Pathologic Anatomy, Univ. of Innsbruck (Austria)

    2003-07-01

    Background: Adjuvant radiotherapy has been shown to improve local control in patients with soft tissue sarcoma. Additional brachytherapy represents a means of enhancing the therapeutic ratio, as biological and dosimetric advantage over single external-beam irradiation (EBRT) can be expected. High-dose-rate intraoperative brachytherapy (IOHDR) as a boost therapy should therefore be able to further diminish the rate of local recurrence even when performing marginal resection. There are sparse data on IOHDR using flab applicators as adjuvant boost to EBRT in combination with marginal resection of soft tissue sarcomas. Patients and Methods: Within a period of 8 years, we prospectively studied 39 adult patients treated by marginal resection, IOHDR using the flab technique and EBRT for soft tissue sarcomas. There were 32 high-grade and seven low-grade tumors, 35 were > 5 cm. Mean follow-up was 26 months (range 3-59 months). Results: We could not detect any local recurrences. No treatment-related loss of limb or life occurred. There were no neurologic or vascular complications, all patients maintained functioning extremities as evidenced by a mean musculoskeletal tumor society (MSTS) functional score of 88.5 (70-100). Treatment-related wound morbidity occurred in eleven patients necessitating revision surgery in eight. Metastatic disease developed in seven patients, six of them had died. The 2-year actuarial disease-free survival was 84%. Conclusions: IOHDR using the flab technique in combination with EBRT and marginal resection is an efficient treatment technique leading to optimal local control rates and limited functional impairment. (orig.)

  2. Interactions of highly and low virulent Flavobacterium columnare isolates with gill tissue in carp and rainbow trout.

    Science.gov (United States)

    Declercq, Annelies Maria; Chiers, Koen; Van den Broeck, Wim; Dewulf, Jeroen; Eeckhaut, Venessa; Cornelissen, Maria; Bossier, Peter; Haesebrouck, Freddy; Decostere, Annemie

    2015-03-06

    The interactions of Flavobacterium columnare isolates of different virulence with the gills of carp (Cyprinus carpio L.) and rainbow trout (Oncorhynchus mykiss Walbaum) were investigated. Both fish species were exposed to different high (HV) or low virulence (LV) isolates and sacrificed at seven predetermined times post-challenge. Histopathological and ultrastructural examination of carp and rainbow trout inoculated with the HV-isolate disclosed bacterial invasion and concomitant destruction of the gill tissue, gradually spreading from the filament tips towards the base, with outer membrane vesicles surrounding most bacterial cells. In carp, 5-10% of the fish inoculated with the LV-isolate became moribund and their gill tissue displayed the same features as described for the HV-isolate, albeit to a lesser degree. The bacterial numbers retrieved from the gill tissue were significantly higher for HV- compared to LV-isolate challenged carp and rainbow trout. TUNEL-stained and caspase-3-immunostained gill sections demonstrated significantly higher apoptotic cell counts in carp and rainbow trout challenged with the HV-isolate compared to control animals. Periodic acid-Schiff/alcian blue staining demonstrated a significantly higher total gill goblet cell count for HV- and LV-isolate challenged compared to control carp. Moreover, bacterial clusters were embedded in a neutral matrix while being encased by acid mucins, resembling biofilm formation. Eosinophilic granular cell counts were significantly higher in the HV-isolate compared to LV-isolate inoculated and control carp. The present data indicate a high colonization capacity, and the destructive and apoptotic-promoting features of the HV-isolate, and point towards important dynamic host mucin-F. columnare interactions warranting further research.

  3. Tissue-specific bioaccumulation of human and veterinary antibiotics in bile, plasma, liver and muscle tissues of wild fish from a highly urbanized region.

    Science.gov (United States)

    Zhao, Jian-Liang; Liu, You-Sheng; Liu, Wang-Rong; Jiang, Yu-Xia; Su, Hao-Chang; Zhang, Qian-Qian; Chen, Xiao-Wen; Yang, Yuan-Yuan; Chen, Jun; Liu, Shuang-Shuang; Pan, Chang-Gui; Huang, Guo-Yong; Ying, Guang-Guo

    2015-03-01

    We investigated the bioaccumulation of antibiotics in bile, plasma, liver and muscle tissues of wild fish from four rivers in the Pearl River Delta region. In total, 12 antibiotics were present in at least one type of fish tissues from nine wild fish species in the four rivers. The mean values of log bioaccumulation factors (log BAFs) for the detected antibiotics in fish bile, plasma, liver, and muscle tissues were at the range of 2.06-4.08, 1.85-3.47, 1.41-3.51, and 0.48-2.70, respectively. As the digestion tissues, fish bile, plasma, and liver showed strong bioaccumulation ability for some antibiotics, indicating a different bioaccumulation pattern from hydrophobic organic contaminants. Human health risk assessment based on potential fish consumption indicates that these antibiotics do not appear to pose an appreciable risk to human health. To the best of our knowledge, this is first report of bioaccumulation patterns of antibiotics in wild fish bile and plasma. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Gestational age dependent changes of the fetal brain, liver and adipose tissue fatty acid compositions in a population with high fish intakes

    NARCIS (Netherlands)

    Kuipers, Remko S.; Luxwolda, Martine F.; Offringa, Pieter J.; Boersma, E. Rudy; Dijck-Brouwer, D. A. Janneke; Muskiet, Frits A. J.

    2012-01-01

    Introduction: There are no data on the intrauterine fatty acid (FA) compositions of brain, liver and adipose tissue of infants born to women with high fish intakes. Subjects and methods: We analyzed the brain (n = 18), liver (n = 14) and adipose tissue (n = 11) FA compositions of 20 stillborn

  5. Fluorescence Quenching Nanoprobes Dedicated to In Vivo Photoacoustic Imaging and High-Efficient Tumor Therapy in Deep-Seated Tissue.

    Science.gov (United States)

    Qin, Huan; Zhou, Ting; Yang, Sihua; Xing, Da

    2015-06-10

    Photoacoustic imaging (PAI) and photoacoustic (PA) therapy have promising applications for treating tumors. It is known that the utilization of high-absorption-coefficient probes can selectively enhance the PAI target contrast and PA tumor therapy efficiency in deep-seated tissue. Here, the design of a probe with the highest availability of optical-thermo conversion by using graphene oxide (GO) and dyes via π-π stacking interactions is reported. The GO serves as a base material for loading dyes and quenching dye fluorescence via fluorescence resonance energy transfer (FRET), with the one purpose of maximum of PA efficiency. Experiments verify that the designed fluorescence quenching nanoprobes can produce stronger PA signals than the sum of the separate signals generated in the dye and the GO. Potential applications of the fluorescence quenching nanoprobes are demonstrated, dedicating to enhance PA contrast of targets in deep-seated tissues and tumors in living mice. PA therapy efficiency both in vitro and in vivo by using the fluorescence quenching nanoprobes is found to be higher than with the commonly used PA therapy agents. Taken together, quenching dye fluorescence via FRET will provide a valid means for developing high-efficiency PA probes. Fluorescence quenching nanoprobes are likely to become a promising candidate for deep-seated tumor imaging and therapy. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. High throughput screening for small molecule therapy for Gaucher disease using patient tissue as the source of mutant glucocerebrosidase.

    Directory of Open Access Journals (Sweden)

    Ehud Goldin

    Full Text Available Gaucher disease (GD, the most common lysosomal storage disorder, results from the inherited deficiency of the lysosomal enzyme glucocerebrosidase (GCase. Previously, wildtype GCase was used for high throughput screening (HTS of large collections of compounds to identify small molecule chaperones that could be developed as new therapies for GD. However, the compounds identified from HTS usually showed reduced potency later in confirmatory cell-based assays. An alternate strategy is to perform HTS on mutant enzyme to identify different lead compounds, including those enhancing mutant enzyme activities. We developed a new screening assay using enzyme extract prepared from the spleen of a patient with Gaucher disease with genotype N370S/N370S. In tissue extracts, GCase is in a more native physiological environment, and is present with the native activator saposin C and other potential cofactors. Using this assay, we screened a library of 250,000 compounds and identified novel modulators of mutant GCase including 14 new lead inhibitors and 30 lead activators. The activities of some of the primary hits were confirmed in subsequent cell-based assays using patient-derived fibroblasts. These results suggest that primary screening assays using enzyme extracted from tissues is an alternative approach to identify high quality, physiologically relevant lead compounds for drug development.

  7. Stevioside ameliorates high-fat diet-induced insulin resistance and adipose tissue inflammation by downregulating the NF-{kappa}B pathway

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Zhiquan; Xue, Liqiong; Guo, Cuicui; Han, Bing; Pan, Chunming; Zhao, Shuangxia; Song, Huaidong [State Key Laboratory of Medical Genomics, Shanghai Institute of Endocrine and Metabolic Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025 (China); Ma, Qinyun, E-mail: qinyunma@126.com [State Key Laboratory of Medical Genomics, Shanghai Institute of Endocrine and Metabolic Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025 (China)

    2012-01-27

    Highlights: Black-Right-Pointing-Pointer Stevioside ameliorates high-fat diet-induced insulin resistance. Black-Right-Pointing-Pointer Stevioside alleviates the adipose tissue inflammation. Black-Right-Pointing-Pointer Stevioside reduces macrophages infiltration into the adipose tissue. Black-Right-Pointing-Pointer Stevioside suppresses the activation of NF-{kappa}B in the adipose tissue. -- Abstract: Accumulating evidence suggests that adipose tissue is the main source of pro-inflammatory molecules that predispose individuals to insulin resistance. Stevioside (SVS) is a widely used sweetener with multiple beneficial effects for diabetic patients. In this study, we investigated the effect of SVS on insulin resistance and the pro-inflammatory state of adipose tissue in mice fed with a high-fat diet (HFD). Oral administration of SVS for 1 month had no effect on body weight, but it significantly improved fasting glucose, basal insulin levels, glucose tolerance and whole body insulin sensitivity. Interestingly, these changes were accompanied with decreased expression levels of several inflammatory cytokines in adipose tissue, including TNF-{alpha}, IL6, IL10, IL1{beta}, KC, MIP-1{alpha}, CD11b and CD14. Moreover, macrophage infiltration in adipose tissue was remarkably reduced by SVS. Finally, SVS significantly suppressed the nuclear factor-kappa b (NF-{kappa}B) signaling pathway in adipose tissue. Collectively, these results suggested that SVS may ameliorate insulin resistance in HFD-fed mice by attenuating adipose tissue inflammation and inhibiting the NF-{kappa}B pathway.

  8. Usual and unusual histologic patterns of high Gleason score 8 to 10 adenocarcinoma of the prostate in needle biopsy tissue.

    Science.gov (United States)

    Gottipati, Srinivas; Warncke, Jason; Vollmer, Robin; Humphrey, Peter A

    2012-06-01

    High Gleason score 8 to 10 adenocarcinoma is the most aggressive and potentially lethal form of prostate cancer. The 2005 International Society of Urological Pathology (ISUP)-modified Gleason grading scheme defines several gland arrangements of high Gleason grade patterns 4 and 5. The aim of this investigation was to quantitate the frequency of the ISUP-defined high Gleason grade patterns in needle biopsy tissue, to determine the common admixtures and to characterize patterns not presented in the 2005 ISUP report. For patients who underwent radical prostatectomy, we analyzed for association of specific high-grade patterns in needle biopsy with extraprostatic extension in radical prostatectomy tissues. A total of 268 prostate needle biopsy cases with Gleason score of 8 to 10 were examined. A mean of 3.6 patterns (range, 1 to 8) were identified per case and only 12% of cases had a pure single pattern. Ill-defined glands with poorly formed lumina (at 57%) and fused microacinar glands (at 53%) comprised the predominant and most frequently admixed patterns. Single cells and single signet ring cells were present in 53% and 31% of cases, respectively. Additional patterns in order of frequency included cords (35%), cribriform glands (25%), sheets of cells (19%), chains (4%), glomeruloid (3%), comedonecrosis (2%), and hypernephromatoid (1 case=0.3%). Gleason score 8 to 10 carcinomas are typically extensive in needle core tissue, with a mean of 4.4 positive cores (range, 1 to 15 cores) per case. Only 14 cases (5%) had high-grade minimal carcinoma measuring Gleason grade patterns not described in the 2005 ISUP report include single file growth, solid cylinders, and nested patterns. The single file pattern was present in 40% of cases, and the small solid nested pattern was detected in 24% of cases. One case displayed solid cylinders. Only the single file pattern was associated with extraprostatic extension at radical prostatectomy (P=0.005). These results show that the 2005

  9. Synchrotron infrared imaging of advanced glycation endproducts (AGEs) in cardiac tissue from mice fed high glycemic diets

    Science.gov (United States)

    Birarda, Giovanni; Holman, Elizabeth A.; Fu, Shang; Weikel, Karen; Hu, Ping; Blankenberg, Francis G.; Holman, Hoi-Ying; Taylor, Allen

    2015-01-01

    Recent research findings correlate an increased risk for dieases such as diabetes, macular degeneration and cardiovascular disease (CVD) with diets that rapidly raise the blood sugar levels; these diets are known as high glycemic index (GI) diets which include white breads, sodas and sweet deserts. Lower glycemia diets are usually rich in fruits, non-starchy vegetables and whole grain products. The goal of our study was to compare and contrast the effects of a low vs. high glycemic diet using the biochemical composition and microstructure of the heart. The improved spatial resolution and signal-to-noise for SR-FTIR obtained through the coupling of the bright synchrotron infrared photon source to an infrared spectral microscope enabled the molecular-level observation of diet-related changes within unfixed fresh frozen histologic sections of mouse cardiac tissue. High and low glycemic index (GI) diets were started at the age of five-months and continued for one year, with the diets only differing in their starch distribution (high GI diet = 100% amylopectin versus low GI diet = 30% amylopectin/70% amylose). Serial cryosections of cardiac tissue for SR-FTIR imaging alternated with adjacent hematoxylin and eosin (H&E) stained sections allowed not only fine-scale chemical analyses of glycogen and glycolipid accumulation along a vein as well as protein glycation hotspots co-localizing with collagen cold spots but also the tracking of morphological differences occurring in tandem with these chemical changes. As a result of the bright synchrotron infrared photon source coupling, we were able to provide significant molecular evidence for a positive correlation between protein glycation and collagen degradation in our mouse model. Our results bring a new insight not only to the effects of long-term GI dietary practices of the public but also to the molecular and chemical foundation behind the cardiovascular disease pathogenesis commonly seen in diabetic patients. PMID

  10. Establishment and Application of a Method for High-Risk Human Papillomavirus Genotyping in Cervical Cancer Tissue.

    Science.gov (United States)

    Zou, Ruanmin; Xie, Wangkai; Wang, Huijing; Wang, Jian; Xiao, Lanlan; Ji, Shuhui; Chen, Xiangmin; Zhang, Lifang; Xue, Xiangyang; Chen, Jun

    2016-01-01

    Persistent high-risk HPV infection is a major cause of cervical cancer and E6/E7 genes and the Li gene in the HPV genome are key targets to detect high-risk HPV. This study aims to explore the relationship between cervical lesions and E6/7 by establishing a polymerase chain reaction (PCR) to detect multiplex genes based on HPV EE7 genes. It is hoped that such methods will provide a more reliable method for clinical screening and the prevention of cervical cancer. Based on alignment, specific primers were designed for HPV E6/E7 genes, the sequences of which came from five5 high-risk papillomaviruses that are common in China. This enabled an E6/E7 gene detection method based on multiplex PCR to be established. E6/E7 and Li gene testing were then performed on 65 cervical cancer tissue samples. The gene copy number of HPV E6/E7 genes and the Li gene were detected from different classifications by real-time fluorescence quantitative PCR. Out of the 65 cervical cancer tissue samples, 47 (72.31%) showed positive results in E6/E7 multiplex PCR, 21 (32.31%) showed positive results in the Ll gene PCR, and out of the 219 cervical exfoliate cell samples, 56 (25.57%) showed positive results in E6/E7 multiplex PCR, 21 (13.24%) showed positive results in the L1 gene PCR. There were significant differences (p detection and genotype of five high-risk HPVs.

  11. Progression from high insulin resistance to type 2 diabetes does not entail additional visceral adipose tissue inflammation.

    Directory of Open Access Journals (Sweden)

    Nuria Barbarroja

    Full Text Available Obesity is associated with a low-grade chronic inflammation state. As a consequence, adipose tissue expresses pro-inflammatory cytokines that propagate inflammatory responses systemically elsewhere, promoting whole-body insulin resistance and consequential islet β-cell exhaustation. Thus, insulin resistance is considered the early stage of type 2 diabetes. However, there is evidence of obese individuals that never develop diabetes indicating that the mechanisms governing the association between the increase of inflammatory factors and type 2 diabetes are much more complex and deserve further investigation. We studied for the first time the differences in insulin signalling and inflammatory pathways in blood and visceral adipose tissue (VAT of 20 lean healthy donors and 40 equal morbidly obese (MO patients classified in high insulin resistance (high IR degree and diabetes state. We studied the changes in proinflammatory markers and lipid content from serum; macrophage infiltration, mRNA expression of inflammatory cytokines and transcription factors, activation of kinases involved in inflammation and expression of insulin signalling molecules in VAT. VAT comparison of these experimental groups revealed that type 2 diabetic-MO subjects exhibit the same pro-inflammatory profile than the high IR-MO patients, characterized by elevated levels of IL-1β, IL-6, TNFα, JNK1/2, ERK1/2, STAT3 and NFκB. Our work rules out the assumption that the inflammation should be increased in obese people with type 2 diabetes compared to high IR obese. These findings indicate that some mechanisms, other than systemic and VAT inflammation must be involved in the development of type 2 diabetes in obesity.

  12. Review: two-photon scanning systems for clinical high resolution in vivo tissue imaging

    Science.gov (United States)

    König, K.; Müller, J.; Höfer, M.; Müller, C.; Weinigel, M.; Bückle, R.; Elsner, P.; Kaatz, M.; Messerschmidt, B.

    2008-02-01

    The femtosecond laser multiphoton tomograph DermaInspect as well as high NA two-photon GRIN microendoscopes for in vivo tomography of human skin have been used to detect malignant melanoma as well as to study the diffusion and intradermal accumulation of topically applied cosmetical and pharmaceutical components. So far, more than 500 patients and volunteers in Europe, Australia, and Asia have been investigated with this unique tomograph. Near infrared 80 MHz picojoule femtosecond laser pulses were employed to excite endogenous fluorophores such as NAD(P)H, flavoproteins, melanin, and elastin as well as fluorescent components of a variety of ointments via a twophoton excitation process. In addition, collagen has been imaged by second harmonic generation. Using a two-PMT detection system, the ratio of elastin to collagen was determined during optical sectioning. A high submicron spatial resolution and 50 picosecond temporal resolution was achieved using galvoscan mirrors and piezodriven focusing optics as well as a time-correlated single photon counting module with a fast microchannel plate detector and fast photomultipliers. Individual intratissue cells, mitochondria, melanosomes, and the morphology of the nuclei as well as extracellular matrix elements could be clearly visualized due to molecular imaging and the calculation of fluorescence lifetime images. Nanoparticles and intratissue drugs have been detected non-invasively, in situ and over a period of up to 3 months. In addition, hydration effects and UV effects were studied by monitoring modifications of cellular morphology and autofluorescence. The system was used to observe the diffusion through the stratum corneum and the accumulation and release of functionalized nanoparticles along hair shafts and epidermal ridges. The DermaInspect been also employed to gain information on skin age and wound healing in patients with ulcers. Novel developments include a galvo/piezo-scan driven flexible articulated arm as

  13. Preparation of highly and generally enriched mammalian tissues for solid state NMR

    Energy Technology Data Exchange (ETDEWEB)

    Wong, Veronica Wai Ching; Reid, David G.; Chow, Wing Ying; Rajan, Rakesh [University of Cambridge, Department of Chemistry (United Kingdom); Green, Maggie [University of Cambridge, Central Biomedical Resources, School of Clinical Medicine (United Kingdom); Brooks, Roger A. [University of Cambridge, Department of Trauma and Orthopaedic Surgery, Addenbrooke’s Hospital (United Kingdom); Duer, Melinda J., E-mail: mjd13@cam.ac.uk [University of Cambridge, Department of Chemistry (United Kingdom)

    2015-10-15

    An appreciable level of isotope labelling is essential for future NMR structure elucidation of mammalian biomaterials, which are either poorly expressed, or unexpressable, using micro-organisms. We present a detailed protocol for high level {sup 13}C enrichment even in slow turnover murine biomaterials (fur keratin), using a customized diet supplemented with commercial labelled algal hydrolysate and formulated as a gel to minimize wastage, which female mice consumed during pregnancy and lactation. This procedure produced approximately eightfold higher fur keratin labelling in pups, exposed in utero and throughout life to label, than in adults exposed for the same period, showing both the effectiveness, and necessity, of this approach.

  14. Disruption of myoblast alignment by highly motile rhabdomyosarcoma cell in tissue structure.

    Science.gov (United States)

    Li, Menglu; Nagamori, Eiji; Kino-Oka, Masahiro

    2017-02-01

    Rhabdomyosarcoma (RMS) is a highly malignant tumor type of skeletal muscle origin, hallmarked by local invasion. Interaction between invasive tumor cells and normal cells plays a major role in tumor invasion and metastasis. Culturing tumor cells in a three-dimensional (3D) model can translate tumor malignancy relevant cell-cell interaction. To mimic tumor heterogeneity in vitro, a co-culture system consisting of a malignant embryonal rhabdomyosarcoma (ERMS) cell line RD and a normal human skeletal muscle myoblast (HSMM) cell line was established by cell sheet technology. Various ratios of RDs to HSMMs were employed to understand the quantitative effect on intercellular interactions. Disruption of sheet structure was observed in heterogeneous cell sheets having a low ratio of RDs to HSMMs, whereas homogeneous HSMM or RD sheets maintained intact structure. Deeper exploration of dynamic tumor cell behavior inside HSMM sheets revealed that HSMM cell alignment was disrupted by highly motile RDs. This study demonstrated that RMS cells are capable of compromising their surrounding environment through induced decay of HSMMs alignment in a cell-based 3D system. This suggests that muscle disruption might be a major consequence of RMS cell invasion into muscles, which could be a promising target to preventing tumor invasion. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  15. Alternatively activated macrophages do not synthesize catecholamines or contribute to adipose tissue adaptive thermogenesis.

    Science.gov (United States)

    Fischer, Katrin; Ruiz, Henry H; Jhun, Kevin; Finan, Brian; Oberlin, Douglas J; van der Heide, Verena; Kalinovich, Anastasia V; Petrovic, Natasa; Wolf, Yochai; Clemmensen, Christoffer; Shin, Andrew C; Divanovic, Senad; Brombacher, Frank; Glasmacher, Elke; Keipert, Susanne; Jastroch, Martin; Nagler, Joachim; Schramm, Karl-Werner; Medrikova, Dasa; Collden, Gustav; Woods, Stephen C; Herzig, Stephan; Homann, Dirk; Jung, Steffen; Nedergaard, Jan; Cannon, Barbara; Tschöp, Matthias H; Müller, Timo D; Buettner, Christoph

    2017-05-01

    Adaptive thermogenesis is the process of heat generation in response to cold stimulation. It is under the control of the sympathetic nervous system, whose chief effector is the catecholamine norepinephrine (NE). NE enhances thermogenesis through β3-adrenergic receptors to activate brown adipose tissue and by 'browning' white adipose tissue. Recent studies have reported that alternative activation of macrophages in response to interleukin (IL)-4 stimulation induces the expression of tyrosine hydroxylase (TH), a key enzyme in the catecholamine synthesis pathway, and that this activation provides an alternative source of locally produced catecholamines during the thermogenic process. Here we report that the deletion of Th in hematopoietic cells of adult mice neither alters energy expenditure upon cold exposure nor reduces browning in inguinal adipose tissue. Bone marrow-derived macrophages did not release NE in response to stimulation with IL-4, and conditioned media from IL-4-stimulated macrophages failed to induce expression of thermogenic genes, such as uncoupling protein 1 (Ucp1), in adipocytes cultured with the conditioned media. Furthermore, chronic treatment with IL-4 failed to increase energy expenditure in wild-type, Ucp1 -/- and interleukin-4 receptor-α double-negative (Il4ra -/- ) mice. In agreement with these findings, adipose-tissue-resident macrophages did not express TH. Thus, we conclude that alternatively activated macrophages do not synthesize relevant amounts of catecholamines, and hence, are not likely to have a direct role in adipocyte metabolism or adaptive thermogenesis.

  16. Lsd1 Ablation Triggers Metabolic Reprogramming of Brown Adipose Tissue

    Directory of Open Access Journals (Sweden)

    Delphine Duteil

    2016-10-01

    Full Text Available Previous work indicated that lysine-specific demethylase 1 (Lsd1 can positively regulate the oxidative and thermogenic capacities of white and beige adipocytes. Here we investigate the role of Lsd1 in brown adipose tissue (BAT and find that BAT-selective Lsd1 ablation induces a shift from oxidative to glycolytic metabolism. This shift is associated with downregulation of BAT-specific and upregulation of white adipose tissue (WAT-selective gene expression. This results in the accumulation of di- and triacylglycerides and culminates in a profound whitening of BAT in aged Lsd1-deficient mice. Further studies show that Lsd1 maintains BAT properties via a dual role. It activates BAT-selective gene expression in concert with the transcription factor Nrf1 and represses WAT-selective genes through recruitment of the CoREST complex. In conclusion, our data uncover Lsd1 as a key regulator of gene expression and metabolic function in BAT.

  17. High-SNR multiple T2(*)-contrast magnetic resonance imaging using a robust denoising method based on tissue characteristics.

    Science.gov (United States)

    Eo, Taejoon; Kim, Taeseong; Jun, Yohan; Lee, Hongpyo; Ahn, Sung Soo; Kim, Dong-Hyun; Hwang, Dosik

    2017-06-01

    To develop an effective method that can suppress noise in successive multiecho T 2 (*)-weighted magnetic resonance (MR) brain images while preventing filtering artifacts. For the simulation experiments, we used multiple T 2 -weighted images of an anatomical brain phantom. For in vivo experiments, successive multiecho MR brain images were acquired from five healthy subjects using a multiecho gradient-recalled-echo (MGRE) sequence with a 3T MRI system. Our denoising method is a nonlinear filter whose filtering weights are determined by tissue characteristics among pixels. The similarity of the tissue characteristics is measured based on the l 2 -difference between two temporal decay signals. Both numerical and subjective evaluations were performed in order to compare the effectiveness of our denoising method with those of conventional filters, including Gaussian low-pass filter (LPF), anisotropic diffusion filter (ADF), and bilateral filter. Root-mean-square error (RMSE), signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) were used in the numerical evaluation. Five observers, including one radiologist, assessed the image quality and rated subjective scores in the subjective evaluation. Our denoising method significantly improves RMSE, SNR, and CNR of numerical phantom images, and CNR of in vivo brain images in comparison with conventional filters (P structure conspicuity (8.2 to 9.4 out of 10) and naturalness (9.2 to 9.8 out of 10) among the conventional filters in the subjective evaluation. This study demonstrates that high-SNR multiple T 2 (*)-contrast MR images can be obtained using our denoising method based on tissue characteristics without noticeable artifacts. Evidence level: 2 J. MAGN. RESON. IMAGING 2017;45:1835-1845. © 2016 International Society for Magnetic Resonance in Medicine.

  18. Treating Cutaneous T-cell Lymphoma with Highly Irregular Surfaces with Photon Irradiation Using Rice as Tissue Compensator

    Directory of Open Access Journals (Sweden)

    Lonika eMajithia

    2015-02-01

    Full Text Available Purpose: Cutaneous T-cell lymphoma (CTCL is known to have an excellent response to radiotherapy, an important treatment modality for this disease. In patients with extremity and digit involvement, the irregular surface and depth variations create difficulty in delivering a homogenous dose using electrons. We sought to evaluate photon irradiation with rice packing as tissue equivalence and determine clinical tolerance and response. Materials and Methods: Three consecutive CTCL patients with extensive lower extremity involvement including the digits were treated using external beam photon therapy with rice packing for tissue compensation. The entire foot was treated to 30-40 Gy in 2-3 Gy per fraction using 6 MV photons prescribed to the mid-plane of an indexed box filled with rice in which the foot was placed. Optically stimulated luminescence dosimeter (OSLD was used for dose measurement to determine the dose deposition to the skin surface. Treatment tolerance and response were monitored with clinical evaluation. Results: All patients tolerated the treatment without treatment breaks. Toxicities included grade 3 erythema and desquamation with resolution within 4 weeks. No late toxicities were observed. All four treated sites had partial response (PR by the end of the treatment course. All patients reported improved functionality after treatment, with less pain, drainage, or swelling. No local recurrence has been observed in these patients with a median follow-up time of 14 months. Conclusion: Tissue compensation with rice packing offers a convenient, inexpensive and reproducible method for the treatment of CTCL with highly irregular surfaces.

  19. A Novel High Mechanical Property PLGA Composite Matrix Loaded with Nanodiamond-Phospholipid Compound for Bone Tissue Engineering.

    Science.gov (United States)

    Zhang, Fan; Song, Qingxin; Huang, Xuan; Li, Fengning; Wang, Kun; Tang, Yixing; Hou, Canglong; Shen, Hongxing

    2016-01-20

    A potential bone tissue engineering material was produced from a biodegradable polymer, poly(lactic-co-glycolic acid) (PLGA), loaded with nanodiamond phospholipid compound (NDPC) via physical mixing. On the basis of hydrophobic effects and physical absorption, we modified the original hydrophilic surface of the nanodiamond (NDs) with phospholipids to be amphipathic, forming a typical core-shell structure. The ND-phospholipid weight ratio was optimized to generate sample NDPC50 (i.e., ND-phospholipid weight ratio of 100:50), and NDPC50 was able to be dispersed in a PLGA matrix at up to 20 wt %. Compared to a pure PLGA matrix, the introduction of 10 wt % of NDPC (i.e., sample NDPC50-PF10) resulted in a significant improvement in the material's mechanical and surface properties, including a decrease in the water contact angle from 80 to 55°, an approximately 100% increase in the Young's modulus, and an approximate 550% increase in hardness, thus closely resembling that of human cortical bone. As a novel matrix supporting human osteoblast (hFOB1.19) growth, NDPC50-PFs with different amounts of NDPC50 demonstrated no negative effects on cell proliferation and osteogenic differentiation. Furthermore, we focused on the behaviors of NDPC-PFs implanted into mice for 8 weeks and found that NDPC-PFs induced acceptable immune response and can reduce the rapid biodegradation of PLGA matrix. Our results represent the first in vivo research on ND (or NDPC) as nanofillers in a polymer matrix for bone tissue engineering. The high mechanical properties, good in vitro and in vivo biocompatibility, and increased mineralization capability suggest that biodegradable PLGA composite matrices loaded with NDPC may potentially be useful for a variety of biomedical applications, especially bone tissue engineering.

  20. Lymphocyte subpopulations in high endothelial venules and lymphatic capillaries of bronchus-associated lymphoid tissue (BALT) in the rat.

    Science.gov (United States)

    Otsuki, Y; Ito, Y; Magari, S

    1989-02-01

    The subpopulations of lymphocytes and non-lymphoid cells in high endothelial venules (HEV) and in lymphatic capillaries surrounding lymphoid follicles in bronchus-associated lymphoid tissue (BALT) were examined by electron microscopy after preembedding the tissue and staining with an immunoperoxidase technique. The results were compared with those obtained in gut-associated lymphoid tissue (GALT) reported previously. Monoclonal mouse-anti-rat T cell, IgG, IgM, IgA, and Ia antisera were used. Plasma cells that were reactive to anti-IgG, anti-IgM, and anti-IgA were detected as cells in which the 3',3'-diaminobenzidine tetrahydroxychloride reaction product was localized in rough endoplasmic reticulum and perinuclear spaces but not on plasma membranes. These plasma cells did not occur in either lymphatic capillaries or HEV in BALT as they did in GALT. Cells with surface Ig (sIg cells), T-cell antigen (T cells), and Ia antigen (Ia cells) were present in BALT. T cells were located predominantly in the follicular area opposite the bronchial epithelium; IgM- and IgG-reactive cells were found in the follicular area adjacent to the bronchial epithelium; and IgA-positive cells were found in the lateral part of the area where the T cells were localized (T-cell area). Ia cells were abundant throughout BALT and in moderate numbers in the epithelium. A striking observation was the presence of "nurse-cell"-like structures in the periphery of BALT. The percentages of T, sIgG, sIgM, and sIgA cells in the HEV were 54.7%, 2.4%, 28.9%, and 27.3%, respectively, and in the lymphatic capillaries, 41.2%, 3.8%, 38.2%, and 21.2%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

  1. High temperature requirement A1 and fibronectin: two possible players in placental tissue remodelling

    Directory of Open Access Journals (Sweden)

    G. Tossetta

    2016-11-01

    Full Text Available High temperature requirement A1 (HtrA1 is a secreted protease involved in placental development. Fibronectin (FN is involved in important process such as wound healing, cell adhesion and spreading, growth, migration, and differentiation. The purpose of this study was to analyse the expression patterns of HtrA1 in relationship to FN and to the key growth zones of placenta such as mesenchymal villi as well as cell islands and cell columns. We demonstrated that FN and HtrA1 are localized in the placental key growth zones suggesting a pivotal role in maintaining the balance among the molecules involved in the placental development and differentiation.

  2. Effect of acoustic streaming on tissue heating due to high-intensity focused ultrasound

    CERN Document Server

    Solovchuk, Maxim A; Thiriet, Marc; Lin, Win-Li

    2011-01-01

    The influences of blood vessels and focused location on temperature distribution during high-intensity focused ultrasound (HIFU) ablation of liver tumors is studied. A three-dimensional acoustics-thermal-fluid coupling model is employed to compute the temperature field in the hepatic cancerous region. The model is based on the linear Westervelt and bioheat equations as well as the nonlinear Navier-Stokes equations for the liver parenchyma and blood vessels. The effect of acoustic streaming is also taken into account in the present HIFU simulation study. Different blood vessel diameters and focal point locations were investigated. We found from this three-dimensional numerical study that in large blood vessels both the convective cooling and acoustic streaming can change considerably the temperature field and the thermal lesion near blood vessels. If the blood vessel is located within the beam width, both acoustic streaming and blood flow cooling effects should be taken into account. The predicted temperature ...

  3. A Micro Saddle Coil with Switchable Sensitivity for Local High-Resolution Imaging of Luminal Tissue

    Directory of Open Access Journals (Sweden)

    Tetsuji Dohi

    2016-04-01

    Full Text Available This paper reports on a micro saddle coil for local high-resolution magnetic resonance imaging (MRI fabricated by embedding a flexible coil pattern into a polydimethyilsiloxane (PDMS tube. We can change the sensitivity of the micro coil by deforming the shape of the coil from a saddle-shaped mode to a planar-shaped mode. The inductance, the resistance, and the Q-factor of the coil in the saddle-shaped mode were 2.45 μH, 3.31 Ω, and 39.9, respectively. Those of the planar-shaped mode were 3.07 μH, 3.92 Ω, and 42.9, respectively. In MRI acquired in saddle-shaped mode, a large visible area existed around the coil. Although the sensitive area was considerably reduced in the planar-shaped mode, clear MRI images were obtained. The signal-to-noise ratios (SNR of the saddle-shaped and planar-shaped modes were 194.9 and 505.9, respectively, at voxel size of 2.0 × 2.0 × 2.0 mm3 and 11.7 and 37.4, respectively, at voxel size of 0.5 × 0.5 × 1.0 mm3. The sensitivity of the saddle-shaped and the planar-shaped modes were about 3 times and 10 times higher, respectively, than those of the medical head coil at both voxel sizes. Thus, the micro saddle coil enabled large-area imaging and highly sensitive imaging by switching the shape of the coil.

  4. Comparison of good- and bad-quality cork: application of high-throughput sequencing of phellogenic tissue.

    Science.gov (United States)

    Teixeira, Rita Teresa; Fortes, Ana Margarida; Pinheiro, Carla; Pereira, Helena

    2014-09-01

    Cork is one of the most valuable non-wood forest products and plays an important role in Mediterranean economies. The production of high-quality cork is dependent on both genome and environment, posing constraints on the industry because an ever-growing amount of bad-quality cork (BQC) development has been observed. In order to identify genes responsible for production of cork of superior quality we performed a comparative analysis using the 454 pyrosequencing approach on phellogenic tissue of good- and bad-quality samples. The transcriptional profiling showed a high number of genes differentially expressed (8.48%) from which 78.8% displayed annotation. Genes more highly represented in BQC are involved in DNA synthesis, RNA processing, proteolysis, and transcription factors related to the abiotic stress response. Putative stomatal/lenticular-associated genes which may be responsible for the disadvantageous higher number of lenticular channels in BQC are also more highly represented. BQC also showed an elevated content of free phenolics. On the other hand, good-quality cork (GQC) can be distinguished by highly expressed genes encoding heat-shock proteins. Together the results provide valuable new information about the molecular events leading to cork formation and provide putative biomarkers associated with cork quality that can be useful in breeding programmes. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  5. The histology of human right atrial tissue in patients with high-risk Obstructive Sleep Apnea and underlying cardiovascular disease: A pilot study

    Directory of Open Access Journals (Sweden)

    Erik M. van Oosten

    2015-03-01

    Conclusions: In this pilot study there were no observable histological differences in human right atrial tissue from individuals at high- and low-risk for OSA. Further investigation would be required for more definitive results.

  6. Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis

    Directory of Open Access Journals (Sweden)

    Husted Søren

    2009-09-01

    Full Text Available Abstract Background Quantitative multi-elemental analysis by inductively coupled plasma (ICP spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data. Results We have developed a micro-scaled microwave digestion procedure and optimized it for accurate elemental profiling of plant materials (1-20 mg dry weight. A commercially available 64-position rotor with 5 ml disposable glass vials, originally designed for microwave-based parallel organic synthesis, was used as a platform for the digestion. The novel micro-scaled method was successfully validated by the use of various certified reference materials (CRM with matrices rich in starch, lipid or protein. When the micro-scaled digestion procedure was applied on single rice grains or small batches of Arabidopsis seeds (1 mg, corresponding to approximately 50 seeds, the obtained elemental profiles closely matched those obtained by conventional analysis using digestion in large volume vessels. Accumulated elemental contents derived from separate analyses of rice grain fractions (aleurone, embryo and endosperm closely matched the total content obtained by analysis of the whole rice grain. Conclusion A high-throughput micro-scaled method has been developed which enables digestion of small quantities of plant samples for subsequent elemental profiling by ICP-spectrometry. The method constitutes a valuable tool for screening of mutants and transformants. In addition, the method facilitates studies of the distribution of essential trace elements between and within plant organs which is relevant for, e.g., breeding programmes aiming at

  7. Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis.

    Science.gov (United States)

    Hansen, Thomas H; Laursen, Kristian H; Persson, Daniel P; Pedas, Pai; Husted, Søren; Schjoerring, Jan K

    2009-09-26

    Quantitative multi-elemental analysis by inductively coupled plasma (ICP) spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data. We have developed a micro-scaled microwave digestion procedure and optimized it for accurate elemental profiling of plant materials (1-20 mg dry weight). A commercially available 64-position rotor with 5 ml disposable glass vials, originally designed for microwave-based parallel organic synthesis, was used as a platform for the digestion. The novel micro-scaled method was successfully validated by the use of various certified reference materials (CRM) with matrices rich in starch, lipid or protein. When the micro-scaled digestion procedure was applied on single rice grains or small batches of Arabidopsis seeds (1 mg, corresponding to approximately 50 seeds), the obtained elemental profiles closely matched those obtained by conventional analysis using digestion in large volume vessels. Accumulated elemental contents derived from separate analyses of rice grain fractions (aleurone, embryo and endosperm) closely matched the total content obtained by analysis of the whole rice grain. A high-throughput micro-scaled method has been developed which enables digestion of small quantities of plant samples for subsequent elemental profiling by ICP-spectrometry. The method constitutes a valuable tool for screening of mutants and transformants. In addition, the method facilitates studies of the distribution of essential trace elements between and within plant organs which is relevant for, e.g., breeding programmes aiming at improvement of the micronutrient density in edible

  8. Effects of activator and high-pull headgear combination therapy: skeletal, dentoalveolar, and soft tissue profile changes.

    Science.gov (United States)

    Marşan, Gülnaz

    2007-04-01

    The aim of this study was to evaluate skeletal, dentoalveolar, and soft tissue profile changes with activator and high-pull headgear combination therapy in patients with Class II malocclusions caused by maxillary prognathism and mandibular retrognathism. The subjects, all in the mixed dentition, were selected from a single centre and were divided into two groups: 28 patients were treated with an incisor double capping activator and a high-pull headgear combination appliance (13 girls, 15 boys mean chronological age 11.7 +/- 1.2 years, skeletal age 12.1 +/- 1.4 years) and an untreated group of 28 subjects (14 girls, 14 boys mean chronological mean age 11.9 +/- 1.1 years, skeletal age 12.3 +/- 1.3 years). The skeletal, dentoalveolar, and soft tissue profile changes that occurred were compared on lateral cephalograms taken before treatment (T0) and after 1.1 +/- 0.3 years when the combination appliance was removed (T1). In the control group, the radiographs were obtained at the start (T0) and after an observation period 1.2 +/- 0.4 years (T1). Statistical analysis was undertaken with Wilcoxon's ranked-sum test for intra-group comparisons and differences between groups with t-test and Bonferroni's test at a level of significance of P headgear combination treatment in these growing patients resulted in a correction of the skeletal Class II relationship (ANB -3.4 degrees), a restriction of maxillary growth (SNA -2.0 degrees, OLp-A -2.3 mm), an advancement of the mandibular structures (SNB +2.6 degrees, FH-NPg +2.3 degrees, OLp-B +2.7 mm, OLp-Pg +2.2 mm), an increase in lower face height (ANS-Me +3.9 mm), a correction of the overjet (-5.4 mm), an improvement in overbite (-2.2 mm), uprighting of the maxillary incisors (U1-FH -5.3 degrees, OLp-U1 -2.5 mm), protrusion of the mandibular incisors (IMPA +2.0 degrees, OLp-L1 +2.7 mm), and a correction of the dental Class II malocclusion (OLp-L6 +3.5 mm). The soft tissue profile changes were a correction of facial convexity (G

  9. Highly tissue specific expression of Sphinx supports its male courtship related role in Drosophila melanogaster.

    Directory of Open Access Journals (Sweden)

    Ying Chen

    2011-04-01

    Full Text Available Sphinx is a lineage-specific non-coding RNA gene involved in regulating courtship behavior in Drosophila melanogaster. The 5' flanking region of the gene is conserved across Drosophila species, with the proximal 300 bp being conserved out to D. virilis and a further 600 bp region being conserved amongst the melanogaster subgroup (D. melanogaster, D. simulans, D. sechellia, D. yakuba, and D. erecta. Using a green fluorescence protein transformation system, we demonstrated that a 253 bp region of the highly conserved segment was sufficient to drive sphinx expression in male accessory gland. GFP signals were also observed in brain, wing hairs and leg bristles. An additional ∼800 bp upstream region was able to enhance expression specifically in proboscis, suggesting the existence of enhancer elements. Using anti-GFP staining, we identified putative sphinx expression signal in the brain antennal lobe and inner antennocerebral tract, suggesting that sphinx might be involved in olfactory neuron mediated regulation of male courtship behavior. Whole genome expression profiling of the sphinx knockout mutation identified significant up-regulated gene categories related to accessory gland protein function and odor perception, suggesting sphinx might be a negative regulator of its target genes.

  10. Highly tissue specific expression of Sphinx supports its male courtship related role in Drosophila melanogaster.

    Science.gov (United States)

    Chen, Ying; Dai, Hongzheng; Chen, Sidi; Zhang, Luoying; Long, Manyuan

    2011-04-26

    Sphinx is a lineage-specific non-coding RNA gene involved in regulating courtship behavior in Drosophila melanogaster. The 5' flanking region of the gene is conserved across Drosophila species, with the proximal 300 bp being conserved out to D. virilis and a further 600 bp region being conserved amongst the melanogaster subgroup (D. melanogaster, D. simulans, D. sechellia, D. yakuba, and D. erecta). Using a green fluorescence protein transformation system, we demonstrated that a 253 bp region of the highly conserved segment was sufficient to drive sphinx expression in male accessory gland. GFP signals were also observed in brain, wing hairs and leg bristles. An additional ∼800 bp upstream region was able to enhance expression specifically in proboscis, suggesting the existence of enhancer elements. Using anti-GFP staining, we identified putative sphinx expression signal in the brain antennal lobe and inner antennocerebral tract, suggesting that sphinx might be involved in olfactory neuron mediated regulation of male courtship behavior. Whole genome expression profiling of the sphinx knockout mutation identified significant up-regulated gene categories related to accessory gland protein function and odor perception, suggesting sphinx might be a negative regulator of its target genes.

  11. High-throughput nuclear magnetic resonance metabolomic footprinting for tissue engineering.

    Science.gov (United States)

    Seagle, Christopher; Christie, Megan A; Winnike, Jason H; McClelland, Randall E; Ludlow, John W; O'Connell, Thomas M; Gamcsik, Michael P; MacDonald, Jeffrey M

    2008-06-01

    We report a high-throughput (HTP) nuclear magnetic resonance (NMR) method for analysis of media components and a metabolic schematic to help easily interpret the data. Spin-lattice relaxation values and concentrations were measured for 19 components and 2 internal referencing agents in pure and 2-day conditioned, hormonally defined media from a 3-dimensional (3D) multicoaxial human bioartificial liver (BAL). The (1)H NMR spectral signal-to-noise ratio is 21 for 0.16 mM alanine in medium and is obtained in 12 min using a 400 MHz NMR spectrometer. For comparison, 2D gel cultures and 3D multicoaxial BALs were batch cultured, with medium changed every day for 15 days after inoculation with human liver cells in Matrigel-collagen type 1 gels. Glutamine consumption was higher by day 8 in the BAL than in 2D culture; lactate production was lower through the 15-day culture period. Alanine was the primary amino acid produced and tracked with lactate or urea production. Glucose and pyruvate consumption were similar in the BAL and 2D cultures. NMR analysis permits quality assurance of the bioreactor by identifying contaminants. Ethanol was observed because of a bioreactor membrane "wetting" procedure. A biochemical scheme is presented illustrating bioreactor metabolomic footprint results and demonstrating how this can be translated to modify bioreactor operational parameters or quality assurance issues.

  12. High contributions of sea ice derived carbon in polar bear (Ursus maritimus) tissue

    Science.gov (United States)

    Galicia, Melissa P.; Thiemann, Gregory W.; Belt, Simon T.; Yurkowski, David J.; Dyck, Markus G.

    2018-01-01

    Polar bears (Ursus maritimus) rely upon Arctic sea ice as a physical habitat. Consequently, conservation assessments of polar bears identify the ongoing reduction in sea ice to represent a significant threat to their survival. However, the additional role of sea ice as a potential, indirect, source of energy to bears has been overlooked. Here we used the highly branched isoprenoid lipid biomarker-based index (H-Print) approach in combination with quantitative fatty acid signature analysis to show that sympagic (sea ice-associated), rather than pelagic, carbon contributions dominated the marine component of polar bear diet (72–100%; 99% CI, n = 55), irrespective of differences in diet composition. The lowest mean estimates of sympagic carbon were found in Baffin Bay bears, which were also exposed to the most rapidly increasing open water season. Therefore, our data illustrate that for future Arctic ecosystems that are likely to be characterised by reduced sea ice cover, polar bears will not only be impacted by a change in their physical habitat, but also potentially in the supply of energy to the ecosystems upon which they depend. This data represents the first quantifiable baseline that is critical for the assessment of likely ongoing changes in energy supply to Arctic predators as we move into an increasingly uncertain future for polar ecosystems. PMID:29360849

  13. Fabrication and characterization of highly porous barium titanate based scaffold coated by Gel/HA nanocomposite with high piezoelectric coefficient for bone tissue engineering applications.

    Science.gov (United States)

    Ehterami, Arian; Kazemi, Mansure; Nazari, Bahareh; Saraeian, Payam; Azami, Mahmoud

    2018-03-01

    It is well established that the piezoelectric effect plays an important physiological role in bone growth, remodeling and fracture healing. Barium titanate, as a well-known piezoelectric ceramic, is especially an attractive material as a scaffold for bone tissue engineering applications. In this regard, we tried to fabricate a highly porous barium titanate based scaffolds by foam replication method and polarize them by applying an external electric field. In order to enhance the mechanical and biological properties, polarized/non-polarized scaffolds were coated with gelatin and nanostructured HA and characterized for their morphologies, porosities, piezoelectric and mechanical properties. The results showed that the compressive strength and piezoelectric coefficient of porous scaffolds increased with the increase of sintering temperature. After being coated with Gel/HA nanocomposite, the interconnected porous structure and pore size of the scaffolds almost remain unchanged while the Gel/nHA-coated scaffolds exhibited enhanced compressive strength and elastic modulus compared with the uncoated samples. Also, the effect of polarizing and coating of optimal scaffolds on adhesion, viability, and proliferation of the MG63 osteoblast-like cell line was evaluated by scanning electron microscope (SEM) and MTT assay. The cell culture experiments revealed that developed scaffolds had good biocompatibility and cells were able to adhere, proliferate and migrate into pores of the scaffolds. Furthermore, cell density was significantly higher in the coated scaffolds at all tested time-points. These results indicated that highly porous barium titanate scaffolds coated with Gel/HA nanocomposite has great potential in tissue engineering applications for bone tissue repair and regeneration. Copyright © 2018 Elsevier Ltd. All rights reserved.

  14. Next-Generation Molecular Histology Using Highly Multiplexed Ion Beam Imaging (MIBI) of Breast Cancer Tissue Specimens for Enhanced Clinical Guidance

    Science.gov (United States)

    2015-07-01

    channels specific for each region of interest. Histone H3 and dsDNA are being used as nuclear markers; lamin A/C and nuclear pore complex are...AWARD NUMBER: W81XWH-14-1-0180 TITLE: Next-Generation Molecular Histology Using Highly Multiplexed Ion Beam Imaging (MIBI) of Breast Cancer Tissue...4. TITLE AND SUBTITLE Next-Generation Molecular Histology Using Highly Multiplexed Ion Beam Imaging (MIBI) of Breast Cancer Tissue Specimens for

  15. Tissue non-specific alkaline phosphatase production by human dental pulp stromal cells is enhanced by high density cell culture.

    Science.gov (United States)

    Tomlinson, Matthew J; Dennis, Caitriona; Yang, Xuebin B; Kirkham, Jennifer

    2015-08-01

    The cell surface hydrolase tissue non-specific alkaline phosphatase (TNAP) (also known as MSCA-1) is used to identify a sub-population of bone marrow stromal cells (BMSCs) with high mineralising potential and is found on subsets of cells within the dental pulp. We aim to determine whether TNAP is co-expressed by human dental pulp stromal cells (hDPSCs) alongside a range of BMSC markers, whether this is an active form of the enzyme and the effects of culture duration and cell density on its expression. Cells from primary dental pulp and culture expanded hDPSCs expressed TNAP. Subsequent analyses revealed persistent TNAP expression and co-expression with BMSC markers such as CD73 and CD90. Flow cytometry and biochemical assays showed that increased culture durations and cell densities enhanced TNAP expression by hDPSCs. Arresting the hDPSC cell cycle also increased TNAP expression. These data confirm that TNAP is co-expressed by hDPSCs together with other BMSC markers and show that cell density affects TNAP expression levels. We conclude that TNAP is a potentially useful marker for hDPSC selection especially for uses in mineralised tissue regenerative therapies.

  16. Quantification of anandamide, oleoylethanolamide and palmitoylethanolamide in rodent brain tissue using high performance liquid chromatography–electrospray mass spectroscopy

    Directory of Open Access Journals (Sweden)

    Daniel J. Liput

    2014-08-01

    Full Text Available Reported concentrations for endocannabinoids and related lipids in biological tissues can vary greatly; therefore, methods used to quantify these compounds need to be validated. This report describes a method to quantify anandamide (AEA, oleoylethanolamide (OEA and palmitoylethanolamide (PEA from rodent brain tissue. Analytes were extracted using acetonitrile without further sample clean up, resolved on a C18 reverse-phase column using a gradient mobile phase and detected using electrospray ionization in positive selected ion monitoring mode on a single quadrupole mass spectrometer. The method produced high recovery rates for AEA, OEA and PEA, ranging from 98.1% to 106.2%, 98.5% to 102.2% and 85.4% to 89.5%, respectively. The method resulted in adequate sensitivity with a lower limit of quantification for AEA, OEA and PEA of 1.4 ng/mL, 0.6 ng/mL and 0.5 ng/mL, respectively. The method was reproducible as intraday and interday accuracies and precisions were under 15%. This method was suitable for quantifying AEA, OEA and PEA from rat brain following pharmacological inhibition of fatty acid amide hydrolase.

  17. High accuracy thermal conductivity measurement of aqueous cryoprotective agents and semi-rigid biological tissues using a microfabricated thermal sensor.

    Science.gov (United States)

    Liang, Xin M; Sekar, Praveen K; Zhao, Gang; Zhou, Xiaoming; Shu, Zhiquan; Huang, Zhongping; Ding, Weiping; Zhang, Qingchuan; Gao, Dayong

    2015-05-20

    An improved thermal-needle approach for accurate and fast measurement of thermal conductivity of aqueous and soft biomaterials was developed using microfabricated thermal conductivity sensors. This microscopic measuring device was comprehensively characterized at temperatures from 0 °C to 40 °C. Despite the previous belief, system calibration constant was observed to be highly temperature-dependent. Dynamic thermal conductivity response during cooling (40 °C to -40 °C) was observed using the miniaturized single tip sensor for various concentrations of CPAs, i.e., glycerol, ethylene glycol and dimethyl sulfoxide. Chicken breast, chicken skin, porcine limb, and bovine liver were assayed to investigate the effect of anatomical heterogeneity on thermal conductivity using the arrayed multi-tip sensor at 20 °C. Experimental results revealed distinctive differences in localized thermal conductivity, which suggests the use of approximated or constant property values is expected to bring about results with largely inflated uncertainties when investigating bio-heat transfer mechanisms and/or performing sophisticated thermal modeling with complex biological tissues. Overall, the presented micro thermal sensor with automated data analysis algorithm is a promising approach for direct thermal conductivity measurement of aqueous solutions and soft biomaterials and is of great value to cryopreservation of tissues, hyperthermia or cryogenic, and other thermal-based clinical diagnostics and treatments.

  18. Diosgenin reorganises hyperglycaemia and distorted tissue lipid profile in high-fat diet-streptozotocin-induced diabetic rats.

    Science.gov (United States)

    Naidu, Parim Brahma; Ponmurugan, Ponnusamy; Begum, Mustapha Sabana; Mohan, Karthick; Meriga, Balaji; RavindarNaik, Ramavat; Saravanan, Ganapathy

    2015-12-01

    Diabetes is often connected with significant morbidity, mortality and also has a pivotal role in the development of cardiovascular diseases. Diet intervention, particularly naturaceutical antioxidants have anti-diabetic potential and avert oxidative damage linked with diabetic pathogenesis. The present study investigated the effects of diosgenin, a saponin from fenugreek, on the changes in lipid profile in plasma, liver, heart and brain in high-fat diet-streptozotocin (HFD-STZ)-induced diabetic rats. Diosgenin was administered to HFD-STZ induced diabetic rats by orally at 60 mg kg(-1) body weight for 30 days to assess its effects on body weight gain, glucose, insulin, insulin resistance and cholesterol, triglycerides, free fatty acids and phospholipids in plasma, liver, heart and brain. The levels of body weight, glucose, insulin, insulin resistance, cholesterol, triglycerides, free fatty acids, phospholipids, VLDL-C and LDL-C were increased significantly (P lipid profile in plasma and tissues. The traditional plant fenugreek and its constituents mediate its anti-diabetic potential through mitigating hyperglycaemic status, altering insulin resistance by alleviating metabolic dysregulation of lipid profile in both plasma and tissues. © 2014 Society of Chemical Industry.

  19. Perinatal overnutrition exacerbates adipose tissue inflammation caused by high-fat feeding in C57BL/6J mice.

    Directory of Open Access Journals (Sweden)

    Brandon D Kayser

    Full Text Available Obesity causes white adipose tissue (WAT inflammation and insulin resistance in some, but not all individuals. Here, we used a mouse model of early postnatal overfeeding to determine the role of neonatal nutrition in lifelong WAT inflammation and metabolic dysfunction. C57BL/6J mice were reared in small litters of 3 (SL or normal litters of 7 pups (NL and fed either regular chow or a 60% high fat diet (HFD from 5 to 17 weeks. At weaning, SL mice did not develop WAT inflammation despite increased fat mass, although there was an up-regulation of WAT Arg1 and Tlr4 expression. On HFD, adult SL mice had greater inguinal fat mass compared to NL mice, however both groups showed similar increases in visceral fat depots and adipocyte hypertrophy. Despite the similar levels of visceral adiposity, SL-HFD mice displayed greater impairments in glucose homeostasis and more pronounced hepatic steatosis compared to NL-HFD mice. In addition, WAT from SL mice fed a HFD displayed greater crown-like structure formation, increased M1 macrophages, and higher cytokine gene expression. Together, these data suggest that early postnatal overnutrition may be a critical determinant of fatty liver and insulin resistance in obese adults by programming the inflammatory capacity of adipose tissue.

  20. High accuracy thermal conductivity measurement of aqueous cryoprotective agents and semi-rigid biological tissues using a microfabricated thermal sensor

    Science.gov (United States)

    Liang, Xin M.; Sekar, Praveen K.; Zhao, Gang; Zhou, Xiaoming; Shu, Zhiquan; Huang, Zhongping; Ding, Weiping; Zhang, Qingchuan; Gao, Dayong

    2015-01-01

    An improved thermal-needle approach for accurate and fast measurement of thermal conductivity of aqueous and soft biomaterials was developed using microfabricated thermal conductivity sensors. This microscopic measuring device was comprehensively characterized at temperatures from 0 °C to 40 °C. Despite the previous belief, system calibration constant was observed to be highly temperature-dependent. Dynamic thermal conductivity response during cooling (40 °C to –40 °C) was observed using the miniaturized single tip sensor for various concentrations of CPAs, i.e., glycerol, ethylene glycol and dimethyl sulfoxide. Chicken breast, chicken skin, porcine limb, and bovine liver were assayed to investigate the effect of anatomical heterogeneity on thermal conductivity using the arrayed multi-tip sensor at 20 °C. Experimental results revealed distinctive differences in localized thermal conductivity, which suggests the use of approximated or constant property values is expected to bring about results with largely inflated uncertainties when investigating bio-heat transfer mechanisms and/or performing sophisticated thermal modeling with complex biological tissues. Overall, the presented micro thermal sensor with automated data analysis algorithm is a promising approach for direct thermal conductivity measurement of aqueous solutions and soft biomaterials and is of great value to cryopreservation of tissues, hyperthermia or cryogenic, and other thermal-based clinical diagnostics and treatments. PMID:25993037

  1. {sup 18}F-choline in experimental soft tissue infection assessed with autoradiography and high-resolution PET

    Energy Technology Data Exchange (ETDEWEB)

    Wyss, Matthias T. [PET Center, Division of Nuclear Medicine, University Hospital Zurich, Raemistrasse 100, 8091, Zurich (Switzerland); Center for Radiopharmaceutical Science of ETH, PSI and USZ, Paul Scherrer Institute, Villigen (Switzerland); Weber, Bruno; Spaeth, Nicolas; Buck, Alfred [PET Center, Division of Nuclear Medicine, University Hospital Zurich, Raemistrasse 100, 8091, Zurich (Switzerland); Honer, Michael; Ametamey, Simon M.; Westera, Gerrit [Center for Radiopharmaceutical Science of ETH, PSI and USZ, Paul Scherrer Institute, Villigen (Switzerland); Bode, Beata [Institute of Pathology, University Hospital, Zurich (Switzerland); Kaim, Achim H. [Klinik Im Schachen, Schaenisweg, Aarau (Switzerland)

    2004-03-01

    For each oncological tracer it is important to know the uptake in non-tumorous lesions. The purpose of this study was to measure the accumulation of fluorine-18 choline (FCH), a promising agent for the evaluation of certain tumour types, in infectious tissue. Unilateral thigh muscle abscesses were induced in five rats by intramuscular injection of 0.1 ml of a bacterial suspension (Staphylococcus aureus, 1.2 x 10{sup 9} CFU/ml). In all animals, FCH accumulation was measured with high-resolution positron emission tomography (PET) on day 6. Autoradiography of the abscess and ipsilateral healthy muscle was performed on day 7 (three animals) and day 11 (two animals) and correlated with histology. In addition, {sup 18}F-fluorodeoxyglucose (FDG) PET was performed on day 5. Increased FCH uptake was noted in specific layers of the abscess wall which contained an infiltrate of mainly granulocytes on day 7 and mainly macrophages on day 11. The autoradiographic standardised uptake values in the most active part of the abscess wall were 2.99 on day 7 (n=3) and 4.05 on day 11 (n=2). In healthy muscle the corresponding values were 0.99 and 0.64. The abscesses were clearly visualised on the FCH and FDG PET images. In conclusion, this study demonstrated avid FCH accumulation in inflammatory tissue, which limits the specificity of FCH for tumour detection. Future studies are now needed to determine the degree of this limitation in human cancer patients. (orig.)

  2. Performance of peanut mutants and their offspring generated from mixed high-energy particle field radiation and tissue culture.

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    Wang, J S; Qiao, L X; Zhao, L S; Wang, P; Guo, B T; Liu, L X; Sui, J M

    2015-09-09

    To develop new ways to breed peanut, we irradiated seeds of the Luhua 11 cultivar with a mixed high-energy particle field at different doses. The embryonic leaflets were extracted as explants and incubated on somatic embryo induction medium and then on somatic embryo germination and regeneration medium. After being grafted, the M1-generation plants were transplanted, and seeds from each M1-generation plant were harvested. In the following year, the M2-generation seeds were planted separately. Some M2-generation plants showed distinct character segregation relative to the mutagenic parent in terms of vigor, fertility, plant height, branch number, and pod size and shape. M2-generation plants that had a high pod weight per plant tended to produce M3-generation offspring that also had a high pod weight per plant, much higher than that of the mutagenic parent, Luhua 11. M4-generation seeds varied greatly in quality, and 35 individuals with an increased fat content (>55%) were obtained. Overall, the results indicate that the combination of mutagenesis via mixed high-energy particle field exposure and tissue culture is promising for peanut breeding.

  3. Excess Folic Acid Increases Lipid Storage, Weight Gain, and Adipose Tissue Inflammation in High Fat Diet-Fed Rats

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    Karen B. Kelly

    2016-09-01

    Full Text Available Folic acid intake has increased to high levels in many countries, raising concerns about possible adverse effects, including disturbances to energy and lipid metabolism. Our aim was to investigate the effects of excess folic acid (EFA intake compared to adequate folic acid (AFA intake on metabolic health in a rodent model. We conducted these investigations in the setting of either a 15% energy low fat (LF diet or 60% energy high fat (HF diet. There was no difference in weight gain, fat mass, or glucose tolerance in EFA-fed rats compared to AFA-fed rats when they were fed a LF diet. However, rats fed EFA in combination with a HF diet had significantly greater weight gain and fat mass compared to rats fed AFA (p < 0.05. Gene expression analysis showed increased mRNA levels of peroxisome proliferator-activated receptor γ (PPARγ and some of its target genes in adipose tissue of high fat-excess folic acid (HF-EFA fed rats. Inflammation was increased in HF-EFA fed rats, associated with impaired glucose tolerance compared to high fat-adequate folic acid (HF-AFA fed rats (p < 0.05. In addition, folic acid induced PPARγ expression and triglyceride accumulation in 3T3-L1 cells. Our results suggest that excess folic acid may exacerbate weight gain, fat accumulation, and inflammation caused by consumption of a HF diet.

  4. A PDGFRα-Mediated Switch toward CD9high Adipocyte Progenitors Controls Obesity-Induced Adipose Tissue Fibrosis.

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    Marcelin, Geneviève; Ferreira, Adaliene; Liu, Yuejun; Atlan, Michael; Aron-Wisnewsky, Judith; Pelloux, Véronique; Botbol, Yair; Ambrosini, Marc; Fradet, Magali; Rouault, Christine; Hénégar, Corneliu; Hulot, Jean-Sébastien; Poitou, Christine; Torcivia, Adriana; Nail-Barthelemy, Raphael; Bichet, Jean-Christophe; Gautier, Emmanuel L; Clément, Karine

    2017-03-07

    Obesity-induced white adipose tissue (WAT) fibrosis is believed to accelerate WAT dysfunction. However, the cellular origin of WAT fibrosis remains unclear. Here, we show that adipocyte platelet-derived growth factor receptor-α-positive (PDGFRα+) progenitors adopt a fibrogenic phenotype in obese mice prone to visceral WAT fibrosis. More specifically, a subset of PDGFRα+ cells with high CD9 expression (CD9high) originates pro-fibrotic cells whereas their CD9low counterparts, committed to adipogenesis, are almost completely lost in the fibrotic WAT. PDGFRα pathway activation promotes a phenotypic shift toward PDGFRα+CD9high fibrogenic cells, driving pathological remodeling and altering WAT function in obesity. These findings translated to human obesity as the frequency of CD9high progenitors in omental WAT (oWAT) correlates with oWAT fibrosis level, insulin-resistance severity, and type 2 diabetes. Collectively, our data demonstrate that in addition to representing a WAT adipogenic niche, different PDGFRα+ cell subsets modulate obesity-induced WAT fibrogenesis and are associated with loss of metabolic fitness. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. T-lymphocyte responses to intestinally absorbed antigens can contribute to adipose tissue inflammation and glucose intolerance during high fat feeding.

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    Yuehui Wang

    Full Text Available BACKGROUND: Obesity is associated with inflammation of visceral adipose tissues, which increases the risk for insulin resistance. Animal models suggest that T-lymphocyte infiltration is an important early step, although it is unclear why these cells are attracted. We have recently demonstrated that dietary triglycerides, major components of high fat diets, promote intestinal absorption of a protein antigen (ovalbumin, "OVA". The antigen was partly transported on chylomicrons, which are prominently cleared in adipose tissues. We hypothesized that intestinally absorbed gut antigens may cause T-lymphocyte associated inflammation in adipose tissue. METHODOLOGY/PRINCIPAL FINDINGS: Triglyceride absorption promoted intestinal absorption of OVA into adipose tissue, in a chylomicron-dependent manner. Absorption tended to be higher in mesenteric than subcutaneous adipose tissue, and was lowest in gonadal tissue. OVA immunoreactivity was detected in stromal vascular cells, including endothelial cells. In OVA-sensitized mice, OVA feeding caused marked accumulation of CD3+ and osteopontin+ cells in mesenteric adipose tissue. The accumulating T-lymphocytes were mainly CD4+. As expected, high-fat (60% kCal diets promoted mesenteric adipose tissue inflammation compared to low-fat diets (10% Kcal, as reflected by increased expression of osteopontin and interferon-gamma. Immune responses to dietary OVA further increased diet-induced osteopontin and interferon-gamma expression in mesenteric adipose. Inflammatory gene expression in subcutaneous tissue did not respond significantly to OVA or dietary fat content. Lastly, whereas OVA responses did not significantly affect bodyweight or adiposity, they significantly impaired glucose tolerance. CONCLUSIONS/SIGNIFICANCE: Our results suggest that loss or lack of immunological tolerance to intestinally absorbed T-lymphocyte antigens can contribute to mesenteric adipose tissue inflammation and defective glucose metabolism

  6. Combining limb-sparing surgery with radiation therapy in high-grade soft tissue sarcoma of extremities - Is it effective?

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    Müller, D A; Beltrami, G; Scoccianti, G; Frenos, F; Capanna, R

    2016-07-01

    Limb-sparing surgery in combination with radiation therapy is a well-established treatment for high-grade soft tissue sarcomas of the extremities. But selection of cases and optimal sequence of irradiation and surgery still remain controversial. 769 patients with a high-grade soft tissue sarcoma of the extremities, who underwent a limb-sparing surgery, were retrospectively reviewed. Group 1 (N = 89) was treated with neo-adjuvant radiation therapy, group 2 (N = 315) with adjuvant irradiation and group 3 (N = 365) with surgery alone. After a mean follow up of 45 months 95 local recurrences occurred resulting in a local recurrence-free survival of 83.2% after 5 years and 75.9% after 10 years. Contaminated surgical margins (Odds ratio: 2.42) and previous inadequate surgeries (Odds ratio: 1.89) were identified as risk factors for failed local control. Neo-adjuvant radiation therapy provides the best local recurrence-free rate for 5 years (90.0%), whereas after 10 years (78.3%) adjuvant irradiation showed better local control. The metastatic-free rate was independent from achieved surgical margins (p = 0.179). Group 1 showed the highest rate of revision surgery (9.0%), followed by group 3 (5.5%) and group 2 (4.4%) (p = 0.085). However, the rate of irradiation-correlated side effects was higher in group 2 (15.2%) than in group 1 (11.2%) (p = 0.221). Surgery has to be effective for successful local control and remains the mainstay of the treatment in combination with neo-adjuvant as well as adjuvant irradiation. In really wide or even radical resections the benefit of radiation therapy can be discussed as the irradiation induced side effects are not negligible. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Convolutional neural networks for an automatic classification of prostate tissue slides with high-grade Gleason score

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    Jiménez del Toro, Oscar; Atzori, Manfredo; Otálora, Sebastian; Andersson, Mats; Eurén, Kristian; Hedlund, Martin; Rönnquist, Peter; Müller, Henning

    2017-03-01

    The Gleason grading system was developed for assessing prostate histopathology slides. It is correlated to the outcome and incidence of relapse in prostate cancer. Although this grading is part of a standard protocol performed by pathologists, visual inspection of whole slide images (WSIs) has an inherent subjectivity when evaluated by different pathologists. Computer aided pathology has been proposed to generate an objective and reproducible assessment that can help pathologists in their evaluation of new tissue samples. Deep convolutional neural networks are a promising approach for the automatic classification of histopathology images and can hierarchically learn subtle visual features from the data. However, a large number of manual annotations from pathologists are commonly required to obtain sufficient statistical generalization when training new models that can evaluate the daily generated large amounts of pathology data. A fully automatic approach that detects prostatectomy WSIs with high-grade Gleason score is proposed. We evaluate the performance of various deep learning architectures training them with patches extracted from automatically generated regions-of-interest rather than from manually segmented ones. Relevant parameters for training the deep learning model such as size and number of patches as well as the inclusion or not of data augmentation are compared between the tested deep learning architectures. 235 prostate tissue WSIs with their pathology report from the publicly available TCGA data set were used. An accuracy of 78% was obtained in a balanced set of 46 unseen test images with different Gleason grades in a 2-class decision: high vs. low Gleason grade. Grades 7-8, which represent the boundary decision of the proposed task, were particularly well classified. The method is scalable to larger data sets with straightforward re-training of the model to include data from multiple sources, scanners and acquisition techniques. Automatically

  8. Detection of tissue coagulation by decorrelation of ultrasonic echo signals in cavitation-enhanced high-intensity focused ultrasound treatment.

    Science.gov (United States)

    Yoshizawa, Shin; Matsuura, Keiko; Takagi, Ryo; Yamamoto, Mariko; Umemura, Shin-Ichiro

    2016-01-01

    A noninvasive technique to monitor thermal lesion formation is necessary to ensure the accuracy and safety of high-intensity focused ultrasound (HIFU) treatment. The purpose of this study is to ultrasonically detect the tissue change due to thermal coagulation in the HIFU treatment enhanced by cavitation microbubbles. An ultrasound imaging probe transmitted plane waves at a center frequency of 4.5 MHz. Ultrasonic radio-frequency (RF) echo signals during HIFU exposure at a frequency of 1.2 MHz were acquired. Cross-correlation coefficients were calculated between in-phase and quadrature (IQ) data of two B-mode images with an interval time of 50 and 500 ms for the estimation of the region of cavitation and coagulation, respectively. Pathological examination of the coagulated tissue was also performed to compare with the corresponding ultrasonically detected coagulation region. The distribution of minimum hold cross-correlation coefficient between two sets of IQ data with 50-ms intervals was compared with a pulse inversion (PI) image. The regions with low cross-correlation coefficients approximately corresponded to those with high brightness in the PI image. The regions with low cross-correlation coefficients in 500-ms intervals showed a good agreement with those with significant change in histology. The results show that the regions of coagulation and cavitation could be ultrasonically detected as those with low cross-correlation coefficients between RF frames with certain intervals. This method will contribute to improve the safety and accuracy of the HIFU treatment enhanced by cavitation microbubbles.

  9. Development of collagen/polydopamine complexed matrix as mechanically enhanced and highly biocompatible semi-natural tissue engineering scaffold.

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    Hu, Yang; Dan, Weihua; Xiong, Shanbai; Kang, Yang; Dhinakar, Arvind; Wu, Jun; Gu, Zhipeng

    2017-01-01

    To improve the mechanical properties and biocompatibility of collagen I matrix, a novel and facile strategy was developed to modify porcine acellular dermal matrix (PADM) via dopamine self-polymerization followed by collagen immobilization to enhance the biological, mechanical and physicochemical properties of PADM. Mechanism study indicated that the polymerization of dopamine onto PADM surface could be regulated by controlling the amount of hydrogen bonds forming between phenol hydroxyl (COH) and nitrogen atom (NCO) within collagen fibers of PADM. The investigations of surface interactions between PDA and PADM illustrated that PDA-PADM system yielded better mechanical properties, thermal stability, surface hydrophilicity and the structural integrity of PADM was maintained after dopamine coating. Furthermore, collagen (COL) was immobilized onto the fresh PDA-PADM to fabricate the collagen-PDA-PADM (COL-PDA-PADM) complexed scaffold. The MTT assay and CLSM observation showed that COL-PDA-PADM had better biocompatibility and higher cellular attachment than pure PADM and COL-PADM without dopamine coating, thus demonstrating the efficacy of PDA as the intermediate layer. Meanwhile, the expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) of COL-PDA-PADM were investigated by an in vivo study. The results revealed that COL-PDA-PADM could effectively promote bFGF and VEGF expression, possibly leading to enhancing the dura repairing process. Overall, this work contributed a new insight into the development of a semi-natural tissue engineering scaffold with high biocompatibility and good mechanical properties. Obtaining scaffolds with high biocompatibility and good mechanical properties is still one of the most challenging issues in tissue engineering. To have excellent in vitro and in vivo performance, scaffolds are desired to have similar mechanical and biological properties as the natural extracellular matrix, such as collagen

  10. Brown adipose tissue: research milestones of a potential player in human energy balance and obesity.

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    Zafrir, B

    2013-10-01

    Obesity and diabetes mellitus are worldwide epidemics driven by the disruption in energy balance. In recent years, it was discovered that functional brown adipose tissue (BAT), once thought to exist mainly in infants, is present in adults, and can be detected during cold stimulation, and is associated with decreased adiposity. Brown fat pads were shown to be highly vascularized and metabolically active and on stimulation, they caused enhanced energy expenditure and increased glucose and fatty acid uptake. These observations drew attention to the possibility that nonshivering thermogenesis mediated by activation of BAT might be important in human energy balance and a potential tool to counter obesity. Recent investigations have revealed significant advances in the understanding of the role of BAT-mediated thermogenesis, uncovering essential knowledge on the origin, differentiation, activation, and regulation of BAT in both murine models and humans. In addition to classic BAT depots, transformation of white adipocytes into brown-like adipocytes, and the development of "beige" cells from distinct precursors, were demonstrated in different animal models and resulted in increased thermogenic activity. Several transcription factors, activating proteins, and hormones are increasingly identified as regulating the development and function of both brown-like adipocytes and classic brown fat pads. This review will summarize the evolution of research on BAT in humans, in light of the renewed scientific interest and growing body of evidence showing that recruitment and activation of BAT and browning of white adipose tissue can affect energy expenditure and may be a future feasible target in the treatment of metabolic diseases. © Georg Thieme Verlag KG Stuttgart · New York.

  11. TFH cells accumulate in mucosal tissues of humanized-DRAG mice and are highly permissive to HIV-1

    Science.gov (United States)

    Allam, Atef; Majji, Sai; Peachman, Kristina; Jagodzinski, Linda; Kim, Jiae; Ratto-Kim, Silvia; Wijayalath, Wathsala; Merbah, Melanie; Kim, Jerome H.; Michael, Nelson L.; Alving, Carl R.; Casares, Sofia; Rao, Mangala

    2015-01-01

    CD4+ T follicular helper cells (TFH) in germinal centers are required for maturation of B-cells. While the role of TFH-cells has been studied in blood and lymph nodes of HIV-1 infected individuals, its role in the mucosal tissues has not been investigated. We show that the gut and female reproductive tract (FRT) of humanized DRAG mice have a high level of human lymphocytes and a high frequency of TFH (CXCR5+PD-1++) and precursor-TFH (CXCR5+PD-1+) cells. The majority of TFH-cells expressed CCR5 and CXCR3 and are the most permissive to HIV-1 infection. A single low-dose intravaginal HIV-1 challenge of humanized DRAG mice results in 100% infectivity with accumulation of TFH-cells mainly in the Peyer’s patches and FRT. The novel finding of TFH-cells in the FRT may contribute to the high susceptibility of DRAG mice to HIV-1 infection. This mouse model thus provides new opportunities to study TFH-cells and to evaluate HIV-1 vaccines. PMID:26034905

  12. Breast Tissue Composition and Immunophenotype and Its Relationship with Mammographic Density in Women at High Risk of Breast Cancer.

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    Jia-Min B Pang

    Full Text Available To investigate the cellular and immunophenotypic basis of mammographic density in women at high risk of breast cancer.Mammograms and targeted breast biopsies were accrued from 24 women at high risk of breast cancer. Mammographic density was classified into Wolfe categories and ranked by increasing density. The histological composition and immunophenotypic profile were quantified from digitized haematoxylin and eosin-stained and immunohistochemically-stained (ERα, ERβ, PgR, HER2, Ki-67, and CD31 slides and correlated to mammographic density.Increasing mammographic density was significantly correlated with increased fibrous stroma proportion (rs (22 = 0.5226, p = 0.0088 and significantly inversely associated with adipose tissue proportion (rs (22 = -0.5409, p = 0.0064. Contrary to previous reports, stromal expression of ERα was common (19/20 cases, 95%. There was significantly higher stromal PgR expression in mammographically-dense breasts (p=0.026.The proportion of stroma and fat underlies mammographic density in women at high risk of breast cancer. Increased expression of PgR in the stroma of mammographically dense breasts and frequent and unexpected presence of stromal ERα expression raises the possibility that hormone receptor expression in breast stroma may have a role in mediating the effects of exogenous hormonal therapy on mammographic density.

  13. Brown adipose tissue (BAT) specific vaspin expression is increased after obesogenic diets and cold exposure and linked to acute changes in DNA-methylation.

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    Weiner, Juliane; Rohde, Kerstin; Krause, Kerstin; Zieger, Konstanze; Klöting, Nora; Kralisch, Susan; Kovacs, Peter; Stumvoll, Michael; Blüher, Matthias; Böttcher, Yvonne; Heiker, John T

    2017-06-01

    Several studies have demonstrated anti-diabetic and anti-obesogenic properties of visceral adipose tissue-derived serine protease inhibitor (vaspin) and so evoked its potential use for treatment of obesity-related diseases. The aim of the study was to unravel physiological regulators of vaspin expression and secretion with a particular focus on its role in brown adipose tissue (BAT) biology. We analyzed the effects of obesogenic diets and cold exposure on vaspin expression in liver and white and brown adipose tissue (AT) and plasma levels. Vaspin expression was analyzed in isolated white and brown adipocytes during adipogenesis and in response to adrenergic stimuli. DNA-methylation within the vaspin promoter was analyzed to investigate acute epigenetic changes after cold-exposure in BAT. Our results demonstrate a strong induction of vaspin mRNA and protein expression specifically in BAT of both cold-exposed and high-fat (HF) or high-sugar (HS) fed mice. While obesogenic diets also upregulated hepatic vaspin mRNA levels, cold exposure tended to increase vaspin gene expression of inguinal white adipose tissue (iWAT) depots. Concomitantly, vaspin plasma levels were decreased upon obesogenic or thermogenic triggers. Vaspin expression was increased during adipogenesis but unaffected by sympathetic activation in brown adipocytes. Analysis of vaspin promoter methylation in AT revealed lowest methylation levels in BAT, which were acutely reduced after cold exposure. Our data demonstrate a novel BAT-specific regulation of vaspin gene expression upon physiological stimuli in vivo with acute epigenetic changes that may contribute to cold-induced expression in BAT. We conclude that these findings indicate functional relevance and potentially beneficial effects of vaspin in BAT function.

  14. The Alteration of the Epidermal Basement Membrane Complex of Human Nevus Tissue and Keratinocyte Attachment after High Hydrostatic Pressurization

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    Naoki Morimoto

    2016-01-01

    Full Text Available We previously reported that human nevus tissue was inactivated after high hydrostatic pressure (HHP higher than 200 MPa and that human cultured epidermis (hCE engrafted on the pressurized nevus at 200 MPa but not at 1000 MPa. In this study, we explore the changes to the epidermal basement membrane in detail and elucidate the cause of the difference in hCE engraftment. Nevus specimens of 8 mm in diameter were divided into five groups (control and 100, 200, 500, and 1000 MPa. Immediately after HHP, immunohistochemical staining was performed to detect the presence of laminin-332 and type VII collagen, and the specimens were observed by transmission electron microscopy (TEM. hCE was placed on the pressurized nevus specimens in the 200, 500, and 1000 MPa groups and implanted into the subcutis of nude mice; the specimens were harvested at 14 days after implantation. Then, human keratinocytes were seeded on the pressurized nevus and the attachment was evaluated. The immunohistochemical staining results revealed that the control and 100 MPa, 200 MPa, and 500 MPa groups were positive for type VII collagen and laminin-332 immediately after HHP. TEM showed that, in all of the groups, the lamina densa existed; however, anchoring fibrils were not clearly observed in the 500 or 1000 MPa groups. Although the hCE took in the 200 and 500 MPa groups, keratinocyte attachment was only confirmed in the 200 MPa group. This result indicates that HHP at 200 MPa is preferable for inactivating nevus tissue to allow its reuse for skin reconstruction in the clinical setting.

  15. Effects of high fat diet, ovariectomy, and physical activity on leptin receptor expression in rat brain and white fat tissue.

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    Blažetić, Senka; Labak, Irena; Viljetić, Barbara; Balog, Marta; Vari, Sandor G; Krivošíková, Zora; Gajdoš, Martin; Kramárová, Patrícia; Kebis, Anton; Vuković, Rosemary; Puljak, Livia; Has-Schön, Elizabeta; Heffer, Marija

    2014-06-01

    To evaluate in a rat animal model whether ovariectomy, high fat diet (HFD), and physical activity in the form of running affect leptin receptor (Ob-R) distribution in the brain and white fat tissue compared to sham (Sh) surgery, standard diet (StD), and sedentary conditions. The study included 48 female laboratory Wistar rats (4 weeks old). Following eight weeks of feeding with standard or HFD, rats were subjected to either OVX or Sh surgery. After surgery, all animals continued StD or HFD for the next 10 weeks. During these 10 weeks, ovariectomy and Sh groups were subjected to physical activity or sedentary conditions. Free-floating immunohistochemistry and Western blot methods were carried out to detect Ob-R in the brain and adipose tissue. StD-ovariectomy-sedentary group had a greater number of Ob-R positive neurons in lateral hypothalamic nuclei than StD-Sh-sedentary group. There was no difference in Ob-R positive neurons in arcuatus nuclei between all groups. Ob-R distribution in the barrel cortex was higher in HFD group than in StD group. Ob-R presence in perirenal and subcutaneous fat was decreased in StD-ovariectomy group. HFD and ovariectomy increased Ob-R distribution in lateral hypothalamic nuclei, but there was no effect on arcuatus nuclei. Our results are first to suggest that HFD, ovariectomy, and physical activity affect Ob-R distribution in the barrel cortex, which might be correlated with the role of Ob-R in election of food in rats.

  16. Vaccination reduces macrophage infiltration in bronchus-associated lymphoid tissue in pigs infected with a highly virulent Mycoplasma hyopneumoniae strain

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    Vranckx Katleen

    2012-03-01

    Full Text Available Abstract Background Mycoplasma hyopneumoniae is the causative agent of enzootic pneumonia and is responsible for significant economic losses to the pig industry. To better understand the mode of action of a commercial, adjuvanted, inactivated whole cell vaccine and the influence of diversity on the efficacy of vaccination, we investigated samples from vaccinated and non-vaccinated pigs experimentally infected with either a low (LV or a highly virulent (HV M. hyopneumoniae strain. Non-vaccinated and sham-infected control groups were included. Lung tissue samples collected at 4 and 8 weeks post infection (PI were immunohistochemically tested for the presence of T-lymphocytes, B-lymphocytes and macrophages in the bronchus-associated lymphoid tissue (BALT. The number of M. hyopneumoniae organisms in bronchoalveolar lavage (BAL fluid was determined using quantitative PCR at 4 and 8 weeks PI. Serum antibodies against M. hyopneumoniae were determined at 0, 2, 4, 6 and 8 weeks PI. Results The immunostaining revealed a lower density of macrophages in the BALT of the vaccinated groups compared to the non-vaccinated groups. The highest number of M. hyopneumoniae organisms in the BAL fluid was measured at 4 weeks PI for the HV strain and at 8 weeks PI for the LV strain. Vaccination reduced the number of organisms non-significantly, though for the HV strain the reduction was clinically more relevant than for the LV strain. At the level of the individual pigs, a higher lung lesion score was associated with more M. hyopneumoniae organisms in the lungs and a higher density of the investigated immune cells in the BALT. Conclusions In conclusion, the infiltration of macrophages after infection with M. hyopneumoniae is reduced by vaccination. The M. hyopneumoniae replication in the lungs is also reduced in vaccinated pigs, though the HV strain is inhibited more than the LV strain.

  17. Vaccination reduces macrophage infiltration in bronchus-associated lymphoid tissue in pigs infected with a highly virulent Mycoplasma hyopneumoniae strain.

    Science.gov (United States)

    Vranckx, Katleen; Maes, Dominiek; Marchioro, Silvana B; Villarreal, Iris; Chiers, Koen; Pasmans, Frank; Haesebrouck, Freddy

    2012-03-12

    Mycoplasma hyopneumoniae is the causative agent of enzootic pneumonia and is responsible for significant economic losses to the pig industry. To better understand the mode of action of a commercial, adjuvanted, inactivated whole cell vaccine and the influence of diversity on the efficacy of vaccination, we investigated samples from vaccinated and non-vaccinated pigs experimentally infected with either a low (LV) or a highly virulent (HV) M. hyopneumoniae strain. Non-vaccinated and sham-infected control groups were included. Lung tissue samples collected at 4 and 8 weeks post infection (PI) were immunohistochemically tested for the presence of T-lymphocytes, B-lymphocytes and macrophages in the bronchus-associated lymphoid tissue (BALT). The number of M. hyopneumoniae organisms in bronchoalveolar lavage (BAL) fluid was determined using quantitative PCR at 4 and 8 weeks PI. Serum antibodies against M. hyopneumoniae were determined at 0, 2, 4, 6 and 8 weeks PI. The immunostaining revealed a lower density of macrophages in the BALT of the vaccinated groups compared to the non-vaccinated groups. The highest number of M. hyopneumoniae organisms in the BAL fluid was measured at 4 weeks PI for the HV strain and at 8 weeks PI for the LV strain. Vaccination reduced the number of organisms non-significantly, though for the HV strain the reduction was clinically more relevant than for the LV strain. At the level of the individual pigs, a higher lung lesion score was associated with more M. hyopneumoniae organisms in the lungs and a higher density of the investigated immune cells in the BALT. In conclusion, the infiltration of macrophages after infection with M. hyopneumoniae is reduced by vaccination. The M. hyopneumoniae replication in the lungs is also reduced in vaccinated pigs, though the HV strain is inhibited more than the LV strain.

  18. A highly conserved candidate chemoreceptor expressed in both olfactory and gustatory tissues in the malaria vector Anopheles gambiae.

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    Pitts, R Jason; Fox, A Nicole; Zwiebel, Laurence J

    2004-04-06

    Anopheles gambiae is a highly anthropophilic mosquito responsible for the majority of malaria transmission in Africa. The biting and host preference behavior of this disease vector is largely influenced by its sense of smell, which is presumably facilitated by G protein-coupled receptor signaling [Takken, W. & Knols, B. (1999) Annu. Rev. Entomol. 44, 131-157]. Because of the importance of host preference to the mosquitoes' ability to transmit disease, we have initiated studies intended to elucidate the molecular mechanisms underlying olfaction in An. gambiae. In the course of these studies, we have identified a number of genes potentially involved in signal transduction, including a family of candidate odorant receptors. One of these receptors, encoded by GPRor7 (hereafter referred to as AgOr7), is remarkably similar to an odorant receptor that is expressed broadly in olfactory tissues and has been identified in Drosophila melanogaster and other insects [Krieger, J., Klink, O., Mohl, C., Raming, K. & Breer, H. (2003) J. Comp. Physiol. A 189, 519-526; Vosshall, L. B., Amrein, H., Morozov, P. S., Rzhetsky, A. & Axel, R. (1999) Cell 96, 725-736]. We have observed AgOr7 expression in olfactory and gustatory tissues in adult An. gambiae and during several stages of the mosquitoes' development. Within the female adult peripheral chemosensory system, antiserum against the AgOR7 polypeptide labels most sensilla of the antenna and maxillary palp as well as a subset of proboscis sensilla. Furthermore, AgOR7 antiserum labeling is observed within the larval antenna and maxillary palpus. These results are consistent with a role for AgOr7 in both olfaction and gustation in An. gambiae and raise the possibility that AgOr7 orthologs may also be of general importance to both modalities of chemosensation in other insects.

  19. Fetal development of subcutaneous white adipose tissue is dependent on Zfp423.

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    Shao, Mengle; Hepler, Chelsea; Vishvanath, Lavanya; MacPherson, Karen A; Busbuso, Napoleon C; Gupta, Rana K

    2017-01-01

    Zfp423 is a multi zinc-finger transcription factor expressed in preadipocytes and mature adipocytes in vivo. Our recent work has revealed a critical role for Zfp423 in maintaining the fate of white adipocytes in adult mice through suppression of the beige cell thermogenic gene program; loss of Zfp423 in mature adipocytes of adult mice results in a white-to-beige phenotypic switch. However, the exact requirements of Zfp423 in the fetal stages of early adipose development in vivo have not been clarified. Here, we utilize two models that confer adipose-specific Zfp423 inactivation during fetal adipose development (Adiponectin-Cre; Zfp423loxP/loxP and Adiponectin-rtTA; TRE-Cre; Zfp423loxP/loxP). We assess the impact of fetal adipose Zfp423 deletion on the initial formation of adipose tissue and evaluate the metabolic consequences of challenging these animals with high-fat diet feeding. Deletion of Zfp423 during fetal adipose development results in a different phenotype than is observed when deleting Zfp423 in adipocytes of adult mice. Inactivation of Zfp423 during fetal adipose development results in arrested differentiation, specifically of inguinal white adipocytes, rather than a white-to-beige phenotypic switch that occurs when Zfp423 is inactivated in adult mice. This is likely explained by the observation that adiponectin driven Cre expression is active at an earlier stage of the adipocyte life cycle during fetal subcutaneous adipose development than in adult mice. Upon high-fat diet feeding, obese adipose Zfp423-deficient animals undergo a pathological adipose tissue expansion, associated with ectopic lipid deposition and systemic insulin resistance. Our results reveal that Zfp423 is essential for the terminal differentiation of subcutaneous white adipocytes during fetal adipose tissue development. Moreover, our data highlight the striking adverse effects of pathological subcutaneous adipose tissue remodeling on visceral adipose function and systemic nutrient

  20. Brown-adipose-tissue macrophages control tissue innervation and homeostatic energy expenditure.

    Science.gov (United States)

    Wolf, Yochai; Boura-Halfon, Sigalit; Cortese, Nina; Haimon, Zhana; Sar Shalom, Hadas; Kuperman, Yael; Kalchenko, Vyacheslav; Brandis, Alexander; David, Eyal; Segal-Hayoun, Yifat; Chappell-Maor, Louise; Yaron, Avraham; Jung, Steffen

    2017-06-01

    Tissue macrophages provide immunological defense and contribute to the establishment and maintenance of tissue homeostasis. Here we used constitutive and inducible mutagenesis to delete the nuclear transcription regulator Mecp2 in macrophages. Mice that lacked the gene encoding Mecp2, which is associated with Rett syndrome, in macrophages did not show signs of neurodevelopmental disorder but displayed spontaneous obesity, which was linked to impaired function of brown adipose tissue (BAT). Specifically, mutagenesis of a BAT-resident Cx3Cr1(+) macrophage subpopulation compromised homeostatic thermogenesis but not acute, cold-induced thermogenesis. Mechanistically, malfunction of BAT in pre-obese mice with mutant macrophages was associated with diminished sympathetic innervation and local titers of norepinephrine, which resulted in lower expression of thermogenic factors by adipocytes. Mutant macrophages overexpressed the signaling receptor and ligand PlexinA4, which might contribute to the phenotype by repulsion of sympathetic axons expressing the transmembrane semaphorin Sema6A. Collectively, we report a previously unappreciated homeostatic role for macrophages in the control of tissue innervation. Disruption of this circuit in BAT resulted in metabolic imbalance.

  1. Relationship between the Balance of Hypertrophic/Hyperplastic Adipose Tissue Expansion and the Metabolic Profile in a High Glucocorticoids Model

    Directory of Open Access Journals (Sweden)

    María Guillermina Zubiría

    2016-07-01

    Full Text Available Adipose tissue (AT expansion is the result of two processes: hyperplasia and hypertrophy; and both, directly or indirectly, depend on the adipogenic potential of adipocyte precursor cells (APCs. Glucocorticoids (GCs have a potent stimulatory effect on terminal adipogenesis; while their effects on early stages of adipogenesis are largely unknown. In the present work, we study, in a model of high GC levels, the adipogenic potential of APCs from retroperitoneal AT (RPAT and its relationship with RPAT mass expansion. We employed a model of hyper-adiposity (30- and 60-day-old rats due to high endogenous GC levels induced by neonatal treatment with l-monosodium glutamate (MSG. We found that the RPAT APCs from 30-day-old MSG rats showed an increased adipogenic capacity, depending on the APCs’ competency, but not in their number. Analyses of RPAT adipocyte diameter revealed an increase in cell size, regardless of the rat age, indicating the prevalence of a hypertrophic process. Moreover, functional RPAT alterations worsened in 60-day-old rats, suggesting that the hyperplastic AT expansion found in 30-day-old animals might have a protective role. We conclude that GCs chronic excess affects APCs’ adipogenic capacity, modifying their competency. This change would modulate the hyperplastic/hypertrophic balance determining healthy or unhealthy RPAT expansion and, therefore, its functionality.

  2. Photocurable high internal phase emulsions (HIPEs) containing hydroxyapatite for additive manufacture of tissue engineering scaffolds with multi-scale porosity.

    Science.gov (United States)

    Wang, Ai-Juan; Paterson, Thomas; Owen, Robert; Sherborne, Colin; Dugan, James; Li, Jun-Ming; Claeyssens, Frederik

    2016-10-01

    Porous composites containing hydroxyapatite (HA) were templated from high internal phase emulsions (HIPEs) and were further structured using direct-write UV stereolithography to produce composite scaffolds with multi-scale porosity. FTIR, TGA and SEM analyses confirmed that HA was retained after photocuring and subsequent treatments and was incorporated within the polymerised HIPE (polyHIPE). The addition of HA particles to the polyHIPE caused changes in the mechanical properties of the material. An increase in both the Young's modulus and maximum stress at yield was observed compared with the pure polyHIPE from 1.544±0.231 to 4.614±0.775 and 0.177±0.009 to 0.267±0.034MPa, respectively. Except at very high concentrations, adding HA did not adversely cause the phase separation of the HIPE or the porous microstructure of the resulting polyHIPE. In combination with a photoinitiator, the HIPE emulsion containing HA was investigated as a photocurable resin for stereolithography-based additive manufacturing. The material was readily processable into "woodpile" structures via direct-write UV stereolithography, producing scaffolds with multi-scale porosity which may be useful for medical applications such as tissue engineering. In conclusion, HA was successfully added into polyHIPEs, producing a similar porous structure to that of the pure polyHIPE whilst improving the mechanical performance. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. GPU-accelerated FDTD modeling of radio-frequency field-tissue interactions in high-field MRI.

    Science.gov (United States)

    Chi, Jieru; Liu, Feng; Weber, Ewald; Li, Yu; Crozier, Stuart

    2011-06-01

    The analysis of high-field RF field-tissue interactions requires high-performance finite-difference time-domain (FDTD) computing. Conventional CPU-based FDTD calculations offer limited computing performance in a PC environment. This study presents a graphics processing unit (GPU)-based parallel-computing framework, producing substantially boosted computing efficiency (with a two-order speedup factor) at a PC-level cost. Specific details of implementing the FDTD method on a GPU architecture have been presented and the new computational strategy has been successfully applied to the design of a novel 8-element transceive RF coil system at 9.4 T. Facilitated by the powerful GPU-FDTD computing, the new RF coil array offers optimized fields (averaging 25% improvement in sensitivity, and 20% reduction in loop coupling compared with conventional array structures of the same size) for small animal imaging with a robust RF configuration. The GPU-enabled acceleration paves the way for FDTD to be applied for both detailed forward modeling and inverse design of MRI coils, which were previously impractical.

  4. Profiling of serum and tissue high abundance acute-phase proteins of patients with epithelial and germ line ovarian carcinoma

    Directory of Open Access Journals (Sweden)

    Abdul-Rahman Puteri

    2008-07-01

    Full Text Available Abstract Background Acute-phase response involves the simultaneous altered expression of serum proteins in association to inflammation, infection, injury or malignancy. Studies of the acute-phase response usually involve determination of the levels of individual acute-phase serum proteins. In the present study, the acute-phase response of patients with epithelial (EOCa and germ-line (GOCa ovarian carcinoma was investigated using the gel-based proteomic approach, a technique which allowed the simultaneous assessment of the levels of the acute-phase serum high abundance proteins. Data obtained were validated using ELISA and immunostaining of biopsy samples. Results Enhanced expression of clusterin (CLU, α1-antitrypsin, haptoglobin and leucine rich glycoprotein was detected in all patients. However, the levels of α1-antichymotrypsin (ACT was only enhanced in EOCa patients, while patients with GOCa were typically characterized by elevated levels of ceruloplasmin but lower levels of α2-HS glycoprotein. The enhanced expression of CLU in EOCa and GOCa patients and up-regulated expression of ACT specifically in EOCa patients were confirmed by ELISA. Immunohistochemical staining of biopsy samples of EOCa and GOCa patients demonstrated correlation of the acute-phase protein expression. Conclusion Patients with EOCa and GOCa demonstrated distinctive aberrant expression of serum and tissue high abundance acute-phase proteins compared to negative control women.

  5. A comparison of live tissue training and high-fidelity patient simulator: A pilot study in battlefield trauma training.

    Science.gov (United States)

    Savage, Erin C; Tenn, Catherine; Vartanian, Oshin; Blackler, Kristen; Sullivan-Kwantes, Wendy; Garrett, Michelle; Blais, Ann-Renee; Jarmasz, Jerzy; Peng, Henry; Pannell, Dylan; Tien, Homer C

    2015-10-01

    Trauma procedural and management skills are often learned on live tissue. However, there is increasing pressure to use simulators because their fidelity improves and as ethical concerns increase. We randomized military medical technicians (medics) to training on either simulators or live tissue to learn combat casualty care skills to determine if the choice of modality was associated with differences in skill uptake. Twenty medics were randomized to trauma training using either simulators or live tissue. Medics were trained to perform five combat casualty care tasks (surgical airway, needle decompression, tourniquet application, wound packing, and intraosseous line insertion). We measured skill uptake using a structured assessment tool. The medics also completed exit questionnaires and interviews to determine which modality they preferred. We found no difference between groups trained with live tissue versus simulators in how they completed each combat casualty care skill. However, we did find that the modality of assessment affected the assessment score. Finally, we found that medics preferred trauma training on live tissue because of the fidelity of tissue handling in live tissue models. However, they also felt that training on simulators also provided additional training value. We found no difference in performance between medics trained on simulators versus live tissue models. Even so, medics preferred live tissue training over simulation. However, more studies are required, and future studies need to address the measurement bias of measuring outcomes in the same model on which the study participants are trained. Therapeutic/care management study, level II.

  6. Colloidal templating of highly ordered gelatin methacryloyl-based hydrogel platforms for three-dimensional tissue analogues

    National Research Council Canada - National Science Library

    Bae Hoon Lee; Hitomi Shirahama; Myung Hee Kim; Jae Ho Lee; Nam-joon Cho; Lay Poh Tan

    2017-01-01

    Three-dimensional, protein-based hydrogel scaffolds that successfully mimic in vivo extracellular matrix microenvironments are desirable for tissue engineering and regenerative medicine applications...

  7. Tissue factor pathway inhibitor for prediction of placenta-mediated adverse pregnancy outcomes in high-risk women: AngioPred study.

    Science.gov (United States)

    Di Bartolomeo, Aurélie; Chauleur, Céline; Gris, Jean-Christophe; Chapelle, Céline; Noblot, Edouard; Laporte, Silvy; Raia-Barjat, Tiphaine

    2017-01-01

    The study aimed to evaluate if the rate of tissue factor pathway inhibitor during pregnancy and following delivery could be a predictive factor for placenta-mediated adverse pregnancy outcomes in high-risk women. This was a prospective multicentre cohort study of 200 patients at a high risk of occurrence or recurrence of placenta-mediated adverse pregnancy outcomes conducted between June 2008 and October 2010. Measurements of tissue factor pathway inhibitor resistance (normalized ratio) and tissue factor pathway inhibitor activity were performed for the last 72 patients at 20, 24, 28, 32, and 36 weeks of gestation and during the postpartum period. Overall, 15 patients presented a placenta-mediated adverse pregnancy outcome. There was no difference in normalized tissue factor pathway inhibitor ratios between patients with and without placenta-mediated adverse pregnancy outcomes during pregnancy and in the post-partum period. Patients with placenta-mediated adverse pregnancy outcomes had tissue factor pathway inhibitor activity rates that were significantly higher than those in patients without at as early as 24 weeks of gestation. The same results were observed following delivery. Among high-risk women, the tissue factor pathway inhibitor activity of patients with gestational vascular complications is higher than that in other patients. Hence, these markers could augment a screening strategy that includes an analysis of angiogenic factors as well as clinical and ultrasound imaging with Doppler measurement of the uterine arteries.

  8. Influence of resveratrol on endoplasmic reticulum stress and expression of adipokines in adipose tissues/adipocytes induced by high-calorie diet or palmitic acid.

    Science.gov (United States)

    Chen, Li; Wang, Ting; Chen, Guanjun; Wang, Nuojin; Gui, Li; Dai, Fang; Fang, Zhaohui; Zhang, Qiu; Lu, Yunxia

    2017-03-01

    This study aimed to determine whether resveratrol treatment alleviates endoplasmic reticulum stress and changes the expression of adipokines in adipose tissues and cells. 8-week-old male C57BL/6 mice were fed a high-calorie diet (HCD group) or high-calorie diet supplemented with resveratrol (high-calorie diet  + resveratrol group) for 3 months. Insulin resistance, serum lipids and proinflammatory indices, the size and inflammatory cell infiltration in subcutaneous and visceral adipose tissues were analyzed. The gene expressions of endoplasmic reticulum stress, adipokines, and inflammatory cytokines were determined. The induced mature 3T3-L1 cells were pretreated with resveratrol and then palmitic acid, and the gene expressions of endoplasmic reticulum stress, adipokines, and inflammatory cytokines were determined. Subcutaneous and visceral adipose tissues in the high-calorie diet-fed mice exhibited adipocyte hypertrophy, inflammatory activation, and endoplasmic reticulum stress. Resveratrol alleviated high-calorie diet-induced insulin resistance and endoplasmic reticulum stress, increased expression of SIRT1, and reversed expression of adipokines in varying degrees in both subcutaneous and visceral adipose tissues. The effects of resveratrol on palmitic acid-treated adipocytes were similar to those shown in the tissues. Resveratrol treatment obviously reversed adipocyte hypertrophy and insulin resistance by attenuating endoplasmic reticulum stress and inflammation, thus increasing the expression of SIRT1 and inverting the expression of adipokines in vivo and in vitro.

  9. Tissue damage after single high-dose intraoperative irradiation of the canine liver : Evaluation in time by means of radionuclide imaging and light microscopy

    NARCIS (Netherlands)

    Cromheecke, M; Piers, BA; Beekhuis, H; ter Veen, H; Sluiter, WJ; Hoekstra, HJ

    2000-01-01

    To establish the tolerance of liver tissue to single high-dose intraoperative irradiation, the histopathological changes in the canine liver after single high-dose intraoperative irradiation were investigated by means of radionuclide imaging and light microscopy, Intraoperative irradiation at doses

  10. High-fat diet induces changes in adipose tissue trans-4-oxo-2-nonenal and trans-4-hydroxy-2-nonenal levels in a depot-specific manner.

    Science.gov (United States)

    Long, Eric K; Olson, Dalay M; Bernlohr, David A

    2013-10-01

    Protein carbonylation is the covalent modification of proteins by α,β-unsaturated aldehydes produced by nonenzymatic lipid peroxidation of polyunsaturated fatty acids. The most widely studied aldehyde product of lipid peroxidation, trans-4-hydroxy-2-nonenal (4-HNE), is associated with obesity-induced metabolic dysfunction and has demonstrated reactivity toward key proteins involved in cellular function. However, 4-HNE is only one of many lipid peroxidation products and the lipid aldehyde profile in adipose tissue has not been characterized. To further understand the role of oxidative stress in obesity-induced metabolic dysfunction, a novel LC-MS/MS method was developed to evaluate aldehyde products of lipid peroxidation and applied to the analysis of adipose tissue. 4-HNE and trans-4-oxo-2-nonenal (4-ONE) were the most abundant aldehydes present in adipose tissue. In high fat-fed C57Bl/6J and ob/ob mice the levels of lipid peroxidation products were increased 5- to 11-fold in epididymal adipose, unchanged in brown adipose, but decreased in subcutaneous adipose tissue. Epididymal adipose tissue of high fat-fed mice also exhibited increased levels of proteins modified by 4-HNE and 4-ONE, whereas subcutaneous adipose tissue levels of these modifications were decreased. High fat feeding of C57Bl/6J mice resulted in decreased expression of a number of genes linked to antioxidant biology selectively in epididymal adipose tissue. Moreover, TNFα treatment of 3T3-L1 adipocytes resulted in decreased expression of GSTA4, GPx4, and Prdx3 while upregulating the expression of SOD2. These results suggest that inflammatory cytokines selectively downregulate antioxidant gene expression in visceral adipose tissue, resulting in elevated lipid aldehydes and increased protein carbonylation. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. Tissue thickness estimation for high precision head-tracking using a galvanometric laser scanner - a case study.

    Science.gov (United States)

    Wissel, Tobias; Stüber, Patrick; Wagner, Benjamin; Dürichen, Robert; Bruder, Ralf; Schweikard, Achim; Ernst, Floris

    2014-01-01

    Marker-less optical head-tracking constitutes a comfortable alternative with no exposure to radiation for realtime monitoring in radiation therapy. Supporting information such as tissue thickness has the potential to improve spatial tracking accuracy. Here we study how accurate tissue thickness can be estimated from the near-infrared (NIR) backscatter obtained from laser scans. In a case study, optical data was recorded with a galvanometric laser scanner from three subjects. A tissue ground truth from MRI was robustly matched via customized bite blocks. We show that Gaussian Processes accurately model the relationship between NIR features and tissue thickness. They were able to predict the tissue thickness with less than 0.5 mm root mean square error. Individual scaling factors for all features and an additional incident angle feature had positive effects on this performance.

  12. Corepressor SMRT promotes oxidative phosphorylation in adipose tissue and protects against diet-induced obesity and insulin resistance.

    Science.gov (United States)

    Fang, Sungsoon; Suh, Jae Myoung; Atkins, Annette R; Hong, Suk-Hyun; Leblanc, Mathias; Nofsinger, Russell R; Yu, Ruth T; Downes, Michael; Evans, Ronald M

    2011-02-22

    The ligand-dependent competing actions of nuclear receptor (NR)-associated transcriptional corepressor and coactivator complexes allow for the precise regulation of NR-dependent gene expression in response to both temporal and environmental cues. Here we report the mouse model termed silencing mediator of retinoid and thyroid hormone receptors (SMRT)(mRID1) in which targeted disruption of the first receptor interaction domain (RID) of the nuclear corepressor SMRT disrupts interactions with a subset of NRs and leads to diet-induced superobesity associated with a depressed respiratory exchange ratio, decreased ambulatory activity, and insulin resistance. Although apparently normal when chow fed, SMRT(mRID1) mice develop multiple metabolic dysfunctions when challenged by a high-fat diet, manifested by marked lipid accumulation in white and brown adipose tissue and the liver. The increased weight gain of SMRT(mRID1) mice on a high-fat diet occurs predominantly in fat with adipocyte hypertrophy evident in both visceral and s.c. depots. Importantly, increased inflammatory gene expression was detected only in the visceral depots. SMRT(mRID1) mice are both insulin-insensitive and refractory to the glucose-lowering effects of TZD and AICAR. Increased serum cholesterol and triglyceride levels were observed, accompanied by increased leptin and decreased adiponectin levels. Aberrant storage of lipids in the liver occurred as triglycerides and cholesterol significantly compromised hepatic function. Lipid accumulation in brown adipose tissue was associated with reduced thermogenic capacity and mitochondrial biogenesis. Collectively, these studies highlight the essential role of NR corepressors in maintaining metabolic homeostasis and describe an essential role for SMRT in regulating the progression, severity, and therapeutic outcome of metabolic diseases.

  13. Determination of valnemulin in swine and bovine tissues by ultra-high performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Li, Hui; Wang, Yingyu; Li, Xiaowei; Fu, Qin; Shan, Yawen; Liu, Tianhe; Xia, Xi

    2016-03-01

    A sensitive and reliable method has been developed and validated for the determination of valnemulin in swine and bovine muscle, liver, and kidney using solid-phase extraction (SPE) and ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The tissue samples were extracted with mixture solution of acetonitrile and 0.01mol/L hydrochloric acid, defatted by n-hexane, and further cleaned up using SPE cartridges with polymeric sorbent. Gradient UHPLC separation was performed using an Acquity BEH C18 column with water and acetonitrile as the mobile phase. Multiple reaction monitoring mode of two precursor-product ion transitions for valnemulin was used. Mean recoveries from fortified samples ranged from 93.4 to 104.3% with 3.3-10.7% relative standard deviation. The limit of detection and quantification was 0.2 and 1μg/kg for the analyte, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Endogenous IL-33 is highly expressed in mouse epithelial barrier tissues, lymphoid organs, brain, embryos, and inflamed tissues: in situ analysis using a novel Il-33-LacZ gene trap reporter strain.

    Science.gov (United States)

    Pichery, Mélanie; Mirey, Emilie; Mercier, Pascale; Lefrancais, Emma; Dujardin, Arnaud; Ortega, Nathalie; Girard, Jean-Philippe

    2012-04-01

    IL-33 (previously known as NF from high endothelial venules) is an IL-1 family cytokine that signals through the ST2 receptor and drives cytokine production in mast cells, basophils, eosinophils, invariant NKT and NK cells, Th2 lymphocytes, and type 2 innate immune cells (natural helper cells, nuocytes, and innate helper 2 cells). Little is known about endogenous IL-33; for instance, the cellular sources of IL-33 in mouse tissues have not yet been defined. In this study, we generated an Il-33-LacZ gene trap reporter strain (Il-33(Gt/Gt)) and used this novel tool to analyze expression of endogenous IL-33 in vivo. We found that the Il-33 promoter exhibits constitutive activity in mouse lymphoid organs, epithelial barrier tissues, brain, and embryos. Immunostaining with anti-IL-33 Abs, using Il-33(Gt/Gt) (Il-33-deficient) mice as control, revealed that endogenous IL-33 protein is highly expressed in mouse epithelial barrier tissues, including stratified squamous epithelia from vagina and skin, as well as cuboidal epithelium from lung, stomach, and salivary gland. Constitutive expression of IL-33 was not detected in blood vessels, revealing the existence of species-specific differences between humans and mice. Importantly, IL-33 protein was always localized in the nucleus of producing cells with no evidence for cytoplasmic localization. Finally, strong expression of the Il-33-LacZ reporter was also observed in inflamed tissues, in the liver during LPS-induced endotoxin shock, and in the lung alveoli during papain-induced allergic airway inflammation. Together, our findings support the possibility that IL-33 may function as a nuclear alarmin to alert the innate immune system after injury or infection in epithelial barrier tissues.

  15. Growing in Antarctica, a challenge for white adipose tissue development in Adelie penguin chicks (Pygoscelis adeliae).

    Science.gov (United States)

    Raccurt, Mireille; Baudimont, Fannie; Tirard, Julien; Rey, Benjamin; Moureaux, Elodie; Géloën, Alain; Duchamp, Claude

    2008-11-01

    Rapid growth is of crucial importance for Adélie penguin chicks reared during the short Antarctic summer. It partly depends on the rapid ontogenesis of fat stores that are virtually null at hatching but then develop considerably (x40) within a month to constitute both an isolative layer against cold and an energy store to fuel thermogenic and growth processes. The present study was aimed at identifying by RT-PCR the major transcriptional events that chronologically underlie the morphological transformation of adipocyte precursors into mature adipocytes from hatching to 30 days of age. The peak expression of GATA binding protein 3, a marker of preadipocytes, at day 7 posthatch indicates a key proliferation step, possibly in relation to the expression of C/EBPalpha (C/EBPalpha). High plasma total 3,5,3'-triiodo-l-thyronine (T(3)) levels and high levels of growth hormone receptor transcripts at hatching suggested that growth hormone and T(3) play early activating roles to favor proliferation of preadipocyte precursors. Differentiation and growth of preadipocytes may occur around day 15 in connection with increased abundance of transcripts encoding IGF-1, proliferator-activated receptor-gamma, and C/EBPbeta, gradually leading to functional maturation of metabolic features of adipocytes including lipid uptake and storage (lipoprotein lipase, fatty-acid synthase) and late endocrine functions (adiponectin) by day 30. Present results show a close correlation between adipose tissue development and chick biology and a difference in the scheduled expression of regulatory factors controlling adipogenesis compared with in vitro studies using cell lines emphasizing the importance of in vivo approaches.

  16. Perioperative high-dose-rate interstitial brachytherapy combined with external beam radiation therapy for soft tissue sarcoma.

    Science.gov (United States)

    Sharma, Daya Nand; Deo, S V Suryanarayana; Rath, Goura Kisor; Shukla, Nootan Kumar; Bakhshi, Sameer; Gandhi, Ajeet Kumar; Julka, Pramod Kumar

    2015-01-01

    The aim of our study was to evaluate the role of perioperative high-dose-rate interstitial brachytherapy (PHDRIBT) in combination with external beam radiation therapy (EBRT) in patients with localized soft tissue sarcoma (STS). From year 2004 to 2010, 52 patients with localized STS were treated with wide local excision plus PHDRIBT followed by EBRT. Median size of the tumor was 8 cm (range, 4-19 cm). A single-plane interstitial brachytherapy implant with an average of nine catheters was performed during the surgical resection. The PHDRIBT was started on third postoperative day to deliver a high-dose-rate dose of 16 Gy in four fractions over 2 days using twice-a-day fractionation schedule. After 4 weeks, EBRT was started for a prescription dose of 50 Gy by conventional fractionation. Subsequently, chemotherapy was administered, if indicated as per our institutional policy. Patients were followed up regularly to study local control, survival, and toxicity. At a median followup of 46 months, no patient developed local recurrence, but 12 patients developed distant metastases. The 5-year overall survival and disease-free survival were 67% and 63%, respectively. Main acute toxicity was delayed wound healing observed in 3 patients (5.7%). Commonest late toxicity was chronic skin/subcutaneous fibrosis noticed in 5 patients (9.6%). The PHDRIBT combined with EBRT provides excellent local control and survival rates with acceptable acute and late toxicity in patients with localized STS. Copyright © 2015 American Brachytherapy Society. Published by Elsevier Inc. All rights reserved.

  17. Catalpol ameliorates high-fat diet-induced insulin resistance and adipose tissue inflammation by suppressing the JNK and NF-κB pathways

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Jun, E-mail: hustzhj@hust.edu.cn; Xu, Gang; Ma, Shuai; Li, Fen; Yuan, Miao; Xu, Huibi; Huang, Kaixun

    2015-11-27

    Catalpol, a bioactive component from the root of Rehmannia glutinosa, has been shown to possess hypoglycemic effects in type 2 diabetic animal models, however, the underlying mechanisms remain poorly understood. Here we investigated the effect of catalpol on high-fat diet (HFD)-induced insulin resistance and adipose tissue inflammation in mice. Oral administration of catalpol at 100 mg/kg for 4 weeks had no effect on body weight of HFD-induced obese mice, but it significantly improved fasting glucose and insulin levels, glucose tolerance and insulin tolerance. Moreover, macrophage infiltration into adipose tissue was markedly reduced by catalpol. Intriguingly, catalpol also significantly reduced mRNA expressions of M1 pro-inflammatory cytokines, but increased M2 anti-inflammatory gene expressions in adipose tissue. Concurrently, catalpol significantly suppressed the c-Jun NH2-terminal kinase (JNK) and nuclear factor-kappa B (NF-κB) signaling pathways in adipose tissue. Collectively, these results suggest that catalpol may ameliorate HFD-induced insulin resistance in mice by attenuating adipose tissue inflammation and suppressing the JNK and NF-κB pathways, and thus provide important new insights into the underlying mechanisms of the antidiabetic effect of catalpol. - Highlights: • Catalpol ameliorates high-fat diet (HFD)-induced insulin resistance in mice. • Catalpol reduces adipose tissue macrophage infiltration in HFD-fed mice. • Catalpol regulates M1 and M2 inflammatory gene expression in obese adipose tissue. • Catalpol suppresses the JNK and NF-κB signaling pathways in obese adipose tissue.

  18. Multiplexed lasing in tissues

    Science.gov (United States)

    Chen, Yu-Cheng; Chen, Qiushu; Fan, Xudong

    2017-02-01

    Biolasers are an emerging technology for next generation biochemical detection and clinical applications. Progress has recently been made to achieve lasing from biomolecules and single living cells. Tissues, which consist of cells embedded in extracellular matrix, mimic more closely the actual complex biological environment in a living body and therefore are of more practical significance. Here, we developed a highly versatile tissue laser platform, in which tissues stained with fluorophores are sandwiched in a high-Q Fabry-Pérot microcavity. Distinct lasing emissions from muscle and adipose tissues stained respectively with fluorescein isothiocyanate (FITC) and boron-dipyrromethene (BODIPY), and hybrid muscle/adipose tissue with dual-staining were achieved with a threshold of only 10 μJ/mm2. Additionally, we investigated how tissue structure/geometry, tissue thickness, and staining dye concentration affect the tissue laser. It is further found that, despite large fluorescence spectral overlap between FITC and BODIPY in tissues, their lasing emissions could be clearly distinguished and controlled due to their narrow lasing bands and different lasing thresholds, thus enabling highly multiplexed detection. Our tissue laser platform can be broadly applicable to various types of tissues/diseases. It provides a new tool for a wide range of biological and biomedical applications, such as diagnostics/screening of tissues and identification/monitoring of biological transformations in tissue engineering.

  19. The bile acid chenodeoxycholic acid directly modulates metabolic pathways in white adipose tissue in vitro: insight into how bile acids decrease obesity.

    Science.gov (United States)

    Teodoro, João Soeiro; Rolo, Anabela Pinto; Jarak, Ivana; Palmeira, Carlos Marques; Carvalho, Rui Albuquerque

    2016-10-01

    Obesity is a worldwide epidemic, and associated pathologies, including type 2 diabetes and cardiovascular alterations, are increasingly escalating morbidity and mortality. Despite intensive study, no effective simple treatment for these conditions exists. As such, the need for go-to drugs is serious. Bile acids (BAs) present the possibility of reversing these problems, as various in vivo studies and clinical trials have shown significant effects with regard to weight and obesity reduction, insulin sensitivity restoration and cardiovascular improvements. However, the mechanism of action of BA-induced metabolic improvement has yet to be fully established. The currently most accepted model involves non-shivering thermogenesis for energy waste, but this is disputed. As such, we propose to determine whether the BA chenodeoxycholic acid (CDCA) can exert anti-obesogenic effects in vitro, independent of thermogenic brown adipose tissue activation. By exposing differentiated 3 T3-L1 adipocytes to high glucose and CDCA, we demonstrate that this BA has anti-obesity effects in vitro. Nuclear magnetic resonance spectroscopic analysis of metabolic pathways clearly indicates an improvement in metabolic status, as these cells become more oxidative rather than glycolytic, which may be associated with an increase in fatty acid oxidation. Our work demonstrates that CDCA-induced metabolic alterations occur in white and brown adipocytes and are not totally dependent on endocrine/nervous system signaling, as thought until now. Furthermore, future exploration of the mechanisms behind these effects will undoubtedly reveal interesting targets for clinical modulation. Copyright © 2016 John Wiley & Sons, Ltd.

  20. Adipose Tissue-Specific Deletion of 12/15-Lipoxygenase Protects Mice from the Consequences of a High-Fat Diet

    Directory of Open Access Journals (Sweden)

    Banumathi K. Cole

    2012-01-01

    Full Text Available Type 2 diabetes is associated with obesity, insulin resistance, and inflammation in adipose tissue. 12/15-Lipoxygenase (12/15-LO generates proinflammatory lipid mediators, which induce inflammation in adipose tissue. Therefore we investigated the role of 12/15-LO activity in mouse white adipose tissue in promoting obesity-induced local and systemic inflammatory consequences. We generated a mouse model for fat-specific deletion of 12/15-LO, aP2-Cre; 12/15-LOloxP/loxP, which we call ad-12/15-LO mice, and placed wild-type controls and ad-12/15-LO mice on a high-fat diet for 16 weeks and examined obesity-induced inflammation and insulin resistance. High-fat diet-fed ad-12/15-LO exhibited improved fasting glucose levels and glucose metabolism, and epididymal adipose tissue from these mice exhibited reduced inflammation and macrophage infiltration compared to wild-type mice. Furthermore, fat-specific deletion of 12/15-LO led to decreased peripheral pancreatic islet inflammation with enlarged pancreatic islets when mice were fed the high-fat diet compared to wild-type mice. These results suggest an interesting crosstalk between 12/15-LO expression in adipose tissue and inflammation in pancreatic islets. Therefore, deletion of 12/15-LO in adipose tissue can offer local and systemic protection from obesity-induced consequences, and blocking 12/15-LO activity in adipose tissue may be a novel therapeutic target in the treatment of type 2 diabetes.

  1. The effects of high-fat diets composed of different animal and vegetable fat sources on the health status and tissue lipid profiles of male Japanese quail (

    Directory of Open Access Journals (Sweden)

    Janine Donaldson

    2017-05-01

    Full Text Available Objective The current study aimed to investigate the impact of high-fat diets composed of different animal and vegetable fat sources on serum metabolic health markers in Japanese quail, as well as the overall lipid content and fatty acid profiles of the edible bird tissues following significantly increased dietary lipid supplementation. Methods Fifty seven male quail were divided into six groups and fed either a standard diet or a diet enriched with one of five different fats (22% coconut oil, lard, palm oil, soybean oil, or sunflower oil for 12 weeks. The birds were subjected to an oral glucose tolerance test following the feeding period, after which they were euthanized and blood, liver, breast, and thigh muscle samples collected. Total fat content and fatty acid profiles of the tissue samples, as well as serum uric acid, triglyceride, cholesterol, total protein, albumin, aspartate transaminase, and total bilirubin concentrations were assessed. Results High-fat diet feeding had no significant effects on the glucose tolerance of the birds. Dietary fatty acid profiles of the added fats were reflected in the lipid profiles of both the liver and breast and thigh muscle tissues, indicating successful transfer of dietary fatty acids to the edible bird tissues. The significantly increased level of lipid inclusion in the diets of the quail used in the present study was unsuccessful in increasing the overall lipid content of the edible bird tissues. Serum metabolic health markers in birds on the high-fat diets were not significantly different from those observed in birds on the standard diet. Conclusion Thus, despite the various high-fat diets modifying the fatty acid profile of the birds’ tissues, unlike in most mammals, the birds maintained a normal health status following consumption of the various high-fat diets.

  2. A large thermogenic-methane release event in the SW Barents Sea, during the Last Glacial Maximum. Indications from numerical modelling and seismic reflection data

    Science.gov (United States)

    Anka, Z.; Rodrigues, E.; di Primio, R.; Ostanin, I.; Stoddart, D.; Horsfield, B.

    2011-12-01

    The Barents Sea, located in the Norwegian Artic area, has undergone a series of tectonic, paleoceanographic and paleo-climatic events during the Cenozoic, which most likely have caused the redistribution and leakage of hydrocarbons accumulations (Ohm et al., 2008). (Dimakis et al., 1998). Present-day under-filled accumulations are known to have leaked in the past providing a source of hydrocarbons, mostly thermogenic methane. However, the timing, extent and driving factors for this event are largely unconstrained. We built a 3D basin model of the Hammerfest Basin in the SW Barents Sea, in order to quantify the masses of liquid and gaseous hydrocarbons generated, accumulated and eventually leaked from the reservoirs during the evolution of the basin. Particular emphasis was placed on analysing the fate of leaked volumes within the dynamics of Plio-Quaternary glacial cycles and formation or destabilization of gas hydrate deposits. The model was calibrated with maturity and temperature well data and reconstructs, with large degree of accuracy, the composition and volume of the hydrocarbons, particularly the gaseous phase present in the main reservoirs. Our results predict the development of overpressures in the reservoirs due to the ice loading of the basin during the glacial periods. Pressure fluctuations derived from cyclic loading-unloading during the glacial-interglacial periods reached up to 5 MPa. The under-filled nature of the present-day accumulations would result from leakage events during the episodes of glacial retreat, in the transition from glacial to interglacial periods. Considerations of the gas hydrate stability conditions in the basin during the time span between 1.00Ma and ≈11,500 years indicate that the leaking thermogenic methane was probably trapped as gas hydrate deposits during the glacial events and then released at once upon hydrate destabilisation during the Last Glacial Maximun (LGM). These results are supported by the presence of km

  3. Orexin restores aging-related brown adipose tissue dysfunction in male mice.

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    Sellayah, Dyan; Sikder, Devanjan

    2014-02-01

    The aging process causes an increase in percent body fat, but the mechanism remains unclear. In the present study we examined the impact of aging on brown adipose tissue (BAT) thermogenic activity as potential cause for the increase in adiposity. We show that aging is associated with interscapular BAT morphologic abnormalities and thermogenic dysfunction. In vitro experiments revealed that brown adipocyte differentiation is defective in aged mice. Interscapular brown tissue in aged mice is progressively populated by adipocytes bearing white morphologic characteristics. Aged mice fail to mobilize intracellular fuel reserves from brown adipocytes and exhibit deficiency in homeothermy. Our results suggest a role for orexin (OX) signaling in the regulation of thermogenesis during aging. Brown fat dysfunction and age-related assimilation of fat mass were accelerated in mice in which OX-producing neurons were ablated. Conversely, OX injections in old mice increased multilocular morphology, increased core body temperature, improved cold tolerance, and reduced adiposity. These results argue that BAT can be targeted for interventions to reverse age-associated increase in fat mass.

  4. Optimisation of high-quality total ribonucleic acid isolation from cartilaginous tissues for real-time polymerase chain reaction analysis

    NARCIS (Netherlands)

    Peeters, M.; Huang, C. L.; Vonk, L. A.; Lu, Z. F.; Bank, R. A.; Helder, M. N.; Doulabi, B. Zandieh

    2016-01-01

    Objectives Studies which consider the molecular mechanisms of degeneration and regeneration of cartilaginous tissues are seriously hampered by problematic ribonucleic acid (RNA) isolations due to low cell density and the dense, proteoglycan-rich extracellular matrix of cartilage. Proteoglycans tend

  5. Optimisation of high-quality total ribonucleic acid isolation from cartilaginous tissues for real-time polymerase chain reaction analysis

    NARCIS (Netherlands)

    Peeters, M; Huang, C L; Vonk, L A; Lu, Z F; Bank, R A; Helder, M N; Doulabi, B Zandieh

    2016-01-01

    OBJECTIVES: Studies which consider the molecular mechanisms of degeneration and regeneration of cartilaginous tissues are seriously hampered by problematic ribonucleic acid (RNA) isolations due to low cell density and the dense, proteoglycan-rich extracellular matrix of cartilage. Proteoglycans tend

  6. Inflammation triggers high mobility group box 1 (HMGB1) secretion in adipose tissue, a potential link to obesity.

    Science.gov (United States)

    Gunasekaran, Manoj Kumar; Viranaicken, Wildriss; Girard, Anne-Claire; Festy, Franck; Cesari, Maya; Roche, Regis; Hoareau, Laurence

    2013-10-01

    Low grade inflammation is one of the major metabolic disorders in case of obesity due to variable secretion of adipose derived cytokines called adipokines. Recently the nuclear protein HMGB1 was identified as an inflammatory alarmin in obesity associated diseases. However HMGB1 role in adipose tissue inflammation is not yet studied. The aim of this study was to prove the expression of HMGB1 in human adipose tissue and to assess the levels of expression between normo-weight and obese individuals. Furthermore we determined which type of cells within adipose tissue is involved in HMGB1 production under inflammatory signal. Western-blot was performed on protein lysates from human normo-weight and obese adipose tissue to study the differential HMGB1 expression. Human normo-weight adipose tissue, adipose-derived stromal cells (ASCs) and adipocytes were cultured and stimulated with LPS to induce inflammation. HMGB1, IL-6 and MCP-1 secretion and gene expression were quantified by ELISA and Q-PCR respectively, as well as cell death by LDH assay. HMGB1 translocation during inflammation was tracked down by immunofluorescence in ASCs. HMGB1 was expressed 2-fold more in adipose tissue from obese compared to normo-weight individuals. LPS led to an up-regulation in