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Forskolin IBMX treatment dramatically activates CREs without altering the circadian properties of the SCN. After 4 hours of induction in a tissue culture incubator, luciferase activity was measured using the Dual Glo . = 0 15 μM for PGE2 and. The herpes simplex virus HSV 1 102 0 promoter contains a putative cAMP response element CRE) located at positions 109 68 to 109 60 with respect to the initiation of transcription.

Figure: Luciferase activities were monitored. The Ready To Glow Secreted Luciferase Assay is based on the secreted Metridia luciferase reporter. Statistical analysis and curve fitting was done by. A) b) c) d) a) b .

We fused them to the cDNA of luciferase Fig. GloResponse™ CRE luc2P HEK293 Cell Line - Promega The authors report the characterization of a novel cyclic adenosine monophosphate cAMP responsive luciferase Luc) reporter. 動します しかし 従来. Forskolin stimulation will increase cellular cAMP levels and increase the luciferase signal derived from the CRE Luc The CRE Luc GPCR Reporter Mouse Platform A Novel Cyclic Adenosine Monophosphate Responsive Luciferase.
Aip regulates cAMP signalling GH secretion in GH3 cells C) CRE luciferase activity with AzA suberic acid in Hana3A cells transfected with N terminal Lucy Flag tagged. These results demonstrate the ability to simultaneously detect two distinct pathway activations within the same cells.
Effects of IBMX in combination with forskolin and Lippia extract on CRE luciferase reporter activity. Assay was performed in accordance with the procedure described above. CRE luciferase reporter vector + constitutively expressing Renilla luciferase vector. Cells are lightly trypsinized and seeded into 96 well plate at.

Luciferase constructs bearing multiple copies of a CRE element: in KTC1 cells no induction by forskolin was observed whereas in. CREB Luciferase Reporter HEK293 Stable Cell Line - Signosis The herpes simplex virus HSV 1 α0 promoter contains a putative cAMP response element CRE) located at positions 68 to 60 with respect to the initiation of transcription. Tance is the demonstration that RIP1 CRE Luc does not exhibit the relatively high levels of basal luciferase activity inherent to.


Involvement of cAMP response element binding protein in the. Role of the Cyclic AMP Response Element Binding Complex and.


F I i i d t CRE L li. Simultaneous detection of multiple. Luciferase reporter cell line The GloResponse™ CRE luc2P HEK293 Cell Line is a clonal GloResponse™ CRE luc2P HEK293 cells response to forskolin CRE CREB Reporter Assay Kit - Amsbio Reporter. The CRE Secreted Luciferase Reporter System is designed to monitor the activation of cAMP binding protein CREB) and cAMP mediated signal transduction pathways.
Cis Regulatory Elements and trans Acting Factors - Circulation. The rat pituitary lactotrope cell.

Results were expressed as percentage of the response induced by 10 µM of Forskolin FSK . Lenti X Ready To Glow Secreted Luciferase Reporter System each.

Dependent inhibition of forskolin stimulated luciferase activ . - Diabetes Luciferase. In this report, the authors examined the functionality of this element using 1) luciferase reporter gene assays in.
ATP + D luciferin + O2 oxyluciferin + AMP + PPi + CO2 + light. In this report, the authors examined the functionality of this element using 1) luciferase reporter gene assays in nerve growth factor differentiated. PDE7A activity was monitored by CRE luciferase activity assay.

To discriminate between the effects of NFIL3 CREB in this experiment we combined. These results suggest that PKA is required for GSNO- mediated CREB phosphorylation. Luciferase reporter assays are used widely to investigate cellular signaling pathways and as high throughput screening tools for drug discovery 1 3 . Ready To Glow Secreted Luciferase Assay - SeparationsSeparations CRE. The VP16 CREB fusion protein moderately stimulates transcription from a minimal promoter containing three copies of the MK5 CRE motif.
PGL3 CRE reporter vectors in 3T3 L1 in response to rosiglitazone forskolin respectively. Luciferase reporter gene constructs were cloned to identify novel regulatory regions for the HBV. This cell line is validated for forskolin stimulation and to the treatment with an inhibitor of the cAMP PKA signaling pathway.
Goa was also able to couple the m4 mAChR receptor to inhibition ofAC. - Google Books Result Transfection of the ac- tivated Q205L Goa subunit decreased forskolin stimula- tion of CRE luciferase expression.

Supplier Data; Technical Support; Certificate of. The miniaturized luciferase assay was performed using. HEK293T cells expressing a CRE responsive luciferase reporter were treated with IBMX alone 25 400 μM) and in the presence of Lippia extract 100 μg mL ; Lippia extract 6 25 100 μg mL) with Fsk 100.

CRE luc2P HEK293 Cell Line Technical Bulletin TB362 - Promega. Background subtracted luciferase readout RLU .

Glucagon Like Peptide 1 Stimulates Insulin Gene. Cyclic AMP Response Element Based.
Ordering Information. Luciferase Reporter Gene Assay in Corning® 384 Well. For Gi coupled 5 HT1A receptor CRE luc based assays were usually performed in the presence of forskolin FSK to raise the cAMP. However forskolin concentration dose response studies revealed a 100 fold amplification of the signal measured by luciferase expression compared with direct cAMP accumulation indicating.
In which a consensus cAMP response element CRE) half site. Highlighting new detection capabilities brought about by engineering luciferase genes enzymes substrates. The presence of renilla luciferase alone both renilla luciferase , firefly luciferase is detected by adding reagent mixture s) to a biological sample producing glow luminescence having a duration of at least an hour.

For example forskolin activates CREB , ATF to bind CRE, treatment of cells with cAMP thus initiating transcription of the secretedMetridia luciferase. The action of GLP 1 at 410RIP1 LUC was also reduced by cotransfection with A CREB, a genetically engineered isoform of the CRE binding protein CREB . Comparison of stable CRE luc2P HEK293 and transiently transfected wildtype HEK293 cells on microscaffolds to a forskolin titration.

Phorylation of Ser133 of CREB. Plasmids containing tandem cAMP response elements CRE .

Miniaturization and Automation of a Luciferase. Positive control luciferase RFP vector was successfully achieved but the C EBPβ or. Data are presented as relative light units RLUs . B) Transient transfection assay of 293T cells with IRS2 promoter construct or control somatostatin CRE luciferase plasmid. A mouse model of Rubinstein Taybi syndrome: Defective long term. Brighter solutions The CRE Secreted Luciferase Reporter System is designed to monitor the activation of cAMP binding protein CREB) and cAMP mediated signal transduction pathways. Timecourse experiments showed that cAMP accumulation preceded luciferase expression and that longer incubations 2 h) were required to gain results with the reporter gene system.
Responsible for further transcriptional induction via CREB activators such as PKA and forskolin. NFIL3 and cAMP Response Element Binding Protein Form a. A) Representative. Berberine decreases IBMX- and FSK induced CRE luciferase activity.

The EC50 of Forskolin is 3 3 µM. These cells then were grown in 96 well plates exposed to drug compounds for four hours then stimulated with a suboptimal dose of forskolin which. C) Chromatin immunoprecipitation assay of the IRS2 promoter using CREB 244 antiserum. CRE CREB Driven Up Regulation of Gene Expression. ABBREVIATIONS: cAMP cyclic adenosine monophosphate response element; GPCR, cyclic adenosine monophosphate; CRE G protein- coupled. Sodium butyrate or 10 mmol L forskolin as a positive control) at 378C.

Cells were co transfected with a luciferase. However, forskolin concentration dose response studies revealed a 100 fold amplification of the signal measured by luciferase. Gq effects on circadian properties of CRE activation in SCN cultures are dose- time dependent persist after drug removal.


1A) and luciferase assays were subsequently performed. The CRE Luciferase Reporter cell line is a stably transfected HEK 293 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the cAMP response element CRE .
In producing luminescence from renilla luciferase alone, a reagent is added. 293 CRE Luciferase Host Cell Line - GenScript CRE Inducibility of 293 CRE Luc Cell Line: Unstimulated Control.

- Google Books Result PKA or forskolin stimulates cotransfected CRE luciferase reporter activity. Key Words: RGS; Astrocytoma cells; Forskolin; RT PCR; PKA; CRE. シグナル伝達ガイド - プロメガ.

Cyclase as measured by this CRE uc reporter gene assay. - Duke Luciferase assays: Use a secreted luciferase reporter for promoter studies in bacteria cultured cells, transgenic plants animals. A) Representative time course of the luciferase expression in HEK293T- CRE Luc cells stably expressing the CRE controlled luciferase upon stimulation with 10 m M of forskolin. The role of c ebp beta in regulating ucp1 expression in 3t3 l1 Luciferase coupled reporter vectors were used to monitor N FAT and CRE regulated gene expression in CHO K1 cells 40 000 cells in 96 well plates) that were.

Evaluation of a CRE directed luciferase reporter gene assay as an. Regulation of cAMP mediated gene transcription by wild type and.

In the luciferase assay because forskolin of this concentration usually induces a half saturating response , we treated cells with 10 μM forskolin for 6 hours to induce CRE activity 6 hours is usually the. Cells on Demand™ Transfection Services - Lonza. LTC1 cells, forskolin produced a 9 fold increase in activity. Differential Signaling by CMV encoded Receptors UL33 and R33.
Fig 1B tests GAL4 fusion contructs of full length CREB cDNA on a. Reporter cells were seeded at a density of 2 X l0 cells/ well in a 48 well culture plate 18 20 h prior to assay. Supplemental Information Elevation of the intracellular cAMP level activates cAMP response element binding protein CREB) to bind CRE and induces the expression of luciferase.

Transfected wild type. The initial screen for luciferase expressing founders was pe forskolin luciferase intensity quantitatively correlates with the severity of the induced psoriasis Finally we have. To further examine the role of CREB in the induction of CRE dependent gene expression by.

HeLa cells were transfected with either the eGFP control IBMX , TORC1 44 eGFP expression vectors with either CRE luciferase , NF AT luciferase before stimulation with forskolin , ionomycin PMA D . Calcineurin fos jun. Butyrate promoted luciferase expression from a CRE reporter construct, induced. Induction of CRE activity by forskolin in CRE HEK293 cells.

Using a simple dual luciferase assay the CRE Reporter CRE Reporter A mixture of inducible CRE cells were treated with different doses of forskolin Orphan Receptors - S T Neurobiology Lab - Google Sites receptors a recombinant Luciferase gene under transcriptional control of a cAMP response element CRE LUC . Activator forskolin in. We examined the possibility that the subunit since the components of EGF- , the CRE Prl luciferase reporters would be in- duced in a similar manner in a heterologous cell type forskolin induced cell signaling cas- cades are likely ubiquitous.

Team NEU China Demonstrate . This may suggest that both CRE elements are functional, but that the MK5 CRE cannot be stimulated by forskolin.

Constitutive Signaling of the Human Cytomegalovirus encoded. Novel 3 D micro scaffold platform: A paradigm shift for cell based. PDE4D Cell Based Activity Assay Kit | BPS 60505 | BPS pLVX CRE MetLuc.
CGTCA 1808 1804 bp) was mutated deleted confirms that this CRE site was required for the induction of luciferase activity by forskolin. 631745 Ready To Glow™ CRE Secreted Luciferase Reporter System Each Data Sheet - BPS Bioscience Western blotting. Luciferase Reporter Gene Assay on Human 5 HT Receptor: Which.

- Shan Lab Inventory of Supplemental Information. SuperLight Luciferase Reporter Gene Assay Kits.

The GloResponse™ CRE luc2P HEK293 Cell Line a f) is a clonal derivative of Human. Applying Transfection Technologies to Create Novel Screening. Reporter gene constructs used for transfection were 4 SomCRET81Luc 17] containing the luciferase gene under the control of four repeats of the CRE from the somatostatin promoter and a truncated thymidine.

Forskolin FSK) treatment increased luciferase activity by approximately 5 fold. Our results indicate that CRE luciferase reporter gene is sensitive and reliable to detect the activities of G protein coupled 5 HT receptors. HEK293 cells response to forskolin titration.

Human olfactory receptors deorphanization - ChemCom Automation Improves Performance of the Low Volume. These plasmids FlucP, were introduced into mammalian cells, along with analogous plasmids containing Fluc gene expression was induced by titrated addition of forskolin FSK . 200 µM) in Hana3A. The amino terminal myristoylation site was removed from wild type Gp 2 and Q205L Gia 2 by changing glycine 2 to. Stimulation of luciferase activity by forskolin. HEK293 cells were transiently transfected with CRE luc and β2 adrenergic receptor cDNA in. JEG3 placental cell line.
PKA or forskolin stimulates cotransfected CRE luciferase reporter. This bioluminescent reporter gene assay is extremely sensitive and is especially suitable for quantifying. Luciferase reporter cell line for analysis of cellular responses that result in modulation of CREB activities.
Fold stimulatory effect of forskolin at RIP1 CRE Luc greatly. Forskolin is commonly used to raise the intracellular level of cAMP in cell physiology studies. FSK activates adenylate cyclase, causing an increase of.

50017 at Vrije Universiteit on December 21, Medical Library light CO PPi AMP in oxylucifer Luciferase O luciferin D ATP + + + . Briefly cells plated one day before were transfected. Relative light units RLU . Forskolin treatment of F11 cells transfected with either the EBPRE luciferase construct or the CRE luciferase construct resulted in an induction of luciferase activity showing peak levels at 24 h after stimulation Fig.


CHO K1 CRE Luciferase Parental Cell Line - GenScript K1 CRE Luc cell line was treated with forskolin as indicated. Discovery of Novel Adenylyl Cyclase Inhibitor by Cell Based. Luciferase assays.

A) Cells were stably transfected with a CRE luciferase reporter gene exposed to vehicle alone , drug HT0712 rolipram) for 2 h before stimulation by a suboptimal dose of forskolin. Optimization of a cAMP response element signal.
CAMP dependent production of Luciferase was. GloResponse TM) CRE luc2P HEK293 Cell Line Technical. The CRE cell line is designed to monitor the cAMP PKA. GloResponse™ CRE luc2P HEK293 Cell Line - Promega Corporation A human neuroblastoma cell line was genetically engineered to carry a CRE luciferase reporter" gene, which makes the cells glow yellow when CREB is activated. Data represent means ± SEM for duplicate samples.

Buy CRE cell line MBS668850) product datasheet at MyBioSource, Cell Lines. Activation of cAMP Response Element Mediated Gene Expression. Cells treated with forskolin or DMSO vehicle. Forskolin resensitizes cell receptors by activating the enzyme adenylyl cyclase and increasing cAMP levels.

RNAi mediated CBP knockdown stop the effects of PKA and FSK on the CRE responsive reporter. For example ATF to bind CRE, forskolin activates CREB , treatment of cells with cAMP thus initiating transcription of the secreted Metridia luciferase.

Luciferase Assay of CHO K1 CRE Luc Reporter Cells: 1. Olfr544 or Olfr545 expression vector n 4 .

Synergism between cAMP and PPAR Signalling in the Initiation of. Up regulation of CRE dependent luciferase expression by prostaglandin E2 PGE2) and adenylyl cyclase activator forskolin in HEK293 cells transiently transfected with a CRE luciferase construct. Treating the cells with forskolin, an activator for.

Related to Figure 2. HEK293 cells transiently transfected with a CRE- luciferase construct. After 4 hours of induction in a tissue culture incubator, luciferase activity.
Of the receptors more than tripled CRE mediated expression through the Gαs pathway; 55% of the receptors inhibited by more than 50 ) forskolin induced CRE. Cignal Lenti CRE Reporter An inducible CRE responsive firefly luciferase reporter cells were treated with different doses of forskolin for 6 hours Evaluation of a CRE Directed Luciferase Reporter Gene Assay as an Alternative to Measuring cAMP Accumulation forskolin stimulated luciferase CRE Reporter – HEK293 Cell Line - - ABGENEX : Antibodies. These cells contain a luciferase gene.

Fi- nally luciferase analysis of 5 deletion mutant revealed that 273 to 16 bp of the renin promoter was sufficient to confer complete forskolin stimulation whereas deletion to 130 dele . ば 細胞内のcAMPレベルを調節している受容体の挙動は 一般にcAMP 応答配列 CRE によってルシフェラーゼの転写と連. LucGFP vectors constructed. Induced by incubation overnight with 20 µM forskolin in a humidity controlled incubator at 37 C with 5% CO2

Evaluation of a CRE directed luciferase reporter gene assay as an alternative to measuring cAMP accumulation. When cells were activated by forskolin . Luciferase activity driven by the Oatp2 promoter containing this CRE site was induced in cells cotrans- fected with a. The luciferase activity was determined after the indicated incubation periods mean.

Figure 1 Effect of lithium on forskolin stimulated CRE CREB directed gene transcription. Subunit gene in the. Embryonic Kidney 293 HEK 293 e) cells.
GloResponse™ CRE luc2P HEK293 Cell Line Luciferase reporter. Luciferase activities were monitored with Analyst HT plate reader upon treatment with forskolin CHO K1 CRE Luc cell line was treated with forskolin as indicated Assay was performed in accordance with the procedure described above Data represent means SEM for duplicate samples Assay Procedure Luciferase Assay of CHO K1 CRE Luc Live Cell Reporters, Now with Lentiviral Delivery Deorphanization screening was performed in HEK293T hRTP1S hRTP2 cells with CRE luciferase reporter assay system12.

CRE luciferase reporter construct in a 384 well plate assay. Lenti X DD Cyan Reporter System each. Using TrkB we showed that the assay was sensitive to physiological concentrations of brain derived neuro- trophic factor BDNF) that the signal was sufficiently ro- bust to be suitable for. Supplemental Information A Gq Ca2+ Axis Controls Circuit Level.

PDE4 inhibitors enhance forskolin induced gene expression in human neuroblastoma cells. The double immunocytochem- ical staining revealed nuclear pCREB immunoreactivity in a proportion of pituitary cells Fig.

Gel shift analysis showed specific DNA protein complexes within this region, which were displaced by the somatostatin consensus CRE. Highly Sensitive Multiplex Luciferase Reporter Assays | Thermo.

Use CRE reporter to study Phosphodiesterase PDE) activity: PDEs regulate the intracellular levels of. - Life Sciences After 4 hours of induction, luciferase activity was measured using the Dual Glo® Luciferase Assay System Reagent on the EnSpire Multimodal reader Perkin Elmer .

Luciferase Reporter Secreted Luciferase Assay - Clontech. Herpesvirus quiescence in neuronal cells. Bioluminescent Assays for High Throughput Screening - URMC Luciferase. Differential expression of the protein kinase A regulatory subunit.

Control of Gene Expression by Catecholamines and the. Forskolin treatment. HEK 293 CRE Luciferase Reporter Cell Line | CrownBio caused an inhibition of forskolin stimulated luciferase ex .

Calcineurin activity is required for depolarization induced, CREB. PLVX CRE DD ZsGreen1. GH3 cells were transfected with Aip siRNA or control non targeting NT) siRNA. Finally which depend on the PKA pathway , forskolin induced the expression of RGS2 mRNA in C6 astrocytoma cells CRE transcriptional pathways.


Evaluation of a CRE Directed Luciferase Reporter Gene Assay as. GSNO, we transfected RPaSMC with a CRE luciferase reporter plasmid together with a plasmid encoding. 3 5 μM for forskolin. And did not modulate the NF κB and AP 1 activity as measured by luciferase reporter gene assay.
FSK and CB2 ligands were incubated with cells for 4 5hr at 37oC. Cholinergic Mechanisms: From Molecular Biology to Clinical. After 22 24 hours of transfection, add threefold serial dilution of forskolin in 50 µl of growth medium.


After 24 h cells were stimulated with forskolin 1 M) for additional 6 h then CRE driven luciferase expres- sion was measured. Activation of cAMP Dependent Signaling Pathway Induces Mouse. Herpesvirus Quiescence in Neuronal Cells. Application: Functional Assay.
Cotransfection with A CREB expression plasmid is indicated. Contains the firefly luciferase gene under the control of multimerized cAMP response element CRE .
Butyrate Activates the cAMP Protein Kinase A - Ping Pong by Nucleofection® with a cAMP inducible luciferase reporter construct. ΜM for PGE2 and 3 5. Regulators of homeostatic functions in the liver CRE binding is increased in hepatic inflammation.
Negative Control. Reporter Component A : CRE luciferase reporter vector + constitutively expressing Renilla luciferase vector; 500 µl. GloResponse™ CRE luc2P HEK293 Cell Line is designed for rapid and Luciferase Assay to determine the activity of forskolin as an agonist of the CRE Regulation of PGC 1α expression in skeletal muscle: Comparison of.


And CRE driven transcription by cannabinoid agonists emphasizes. PGL4 29 Promega) subsequently treated with different Forskolin concentrations. Similarly, the activity of Gaussia luciferase expressed from the CRE Gaussia reporter plasmid increased with increasing concentrations of forskolin Figure 3B .
Cells were harvested using Versene and suspended in serum free DMEM F12 assay buffer. Dose response to Forskolin of CHO CRE luciferase cells, blue line] cryopreserved assay ready on 96 well plates in a proprietary LONZA cryoprotective. Detection limit: 4 cells. Cells were washed twice with serum free DMEM before the addi .

Forskolin Induced Stimulation of RGS2 mRNA in C6 Astrocytoma Cells Fig 1: Induction of CRE activity by forskolin in CRE Leeporter™ – HEK293 cells. D) CRE luciferase assay with AzA and SQ22536. Therefore, the results indicate that RIK of PKA is selectively expressed in pancreatic beta cells as compared to alpha cells: this. Can activate cAMP protein kinase A PKA - cAMP response element CRE binding protein CREB) signaling in Caco 2 cells, a model of intestinal.

After 48 h cells were left either untreated , treated with 1 μM forskolin only 10 μM. HEK 293 CRE luc cells Panomics Cat. Expressing a CRE luciferase reporter gene to develop an as- say for monitoring Trk activation in response to their cognate ligands.

Knock down of endogenous rat Aip in GH3 cells increased cAMP driven transcription of CRE luciferase A) and Id1 cAMP target gene B . Functional Coupling of Endogenous Serotonin 5 HT1B) and. The CRE Leeporter™ Luciferase Reporter cell line is a stably transfected HEK 293 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the cAMP response element CRE .

200 µM FSK forskolin 1 µM) , E) CRE luciferase assay with AzA 30 µM SQ22536. In parallel to the changes in CRE luciferase activity the percentage of pCREB ir lac- totrophs was increased by forskolin treatment the forsko . Optimization of a cAMP response element signal pathway reporter. - Karger Publishers CREB by both GSNO and forskolin.
Transcriptional activity of UCP1 promoter in the presence of forskolin in 3T3 L1 cells without stimulating.
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Figure: Luciferase activities were monitored with Analyst HT plate reader upon treatment with forskolin CHO K1 CRE Luc cell line was treated with forskolin as indicated The assay was performed in accordance with GenScript s standard procedures Data represent means SEM for duplicate samples IV References 1 Conway, S, et al Engineered Luciferase Reporter from a Deep. - ACS Publications Methods and Results. The luciferase assay protocol was as follows: 1.

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Harvest HEK CRE luc cells from culture flask using an enzymatic dissociation solution . Seed cells at 2 500 cells well in 10 20* µL volume IMDM + 10 FBS + 0 1 mg mL hygromycin . Induce cells with 2 5 5 µL of 100 µM Forskolin .

CRE Leeporter™ Luciferase Reporter HEK293 Cell.

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- Cyclic AMP response element CRE binding protein CREB) is a transcription factor that regulates and responds to diverse cellular responses, ranging from proliferation, survival, differentiation, stress responses, and. The hygromycin resistant clones were subsequently screened for forskolin induced luciferase activity. Dart NeuroScience LLC - Internal Research - Memory Enhancers cAMP response element CRE) and the CRE binding protein CREB) as an important mechanism of a cAMP dependent gene regulation.

Methods: We investigated the transcriptional activation by forskolin, an activator of the adenylyl cyclase, in chick embryonic cardiomyocytes transfected with a CRE controlled luciferase. CRE Luc GPCR Reporter Mouse Platform | Taconic Biosciences, Inc.

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Taconic Biosciences provides an in vitro in vivo luciferase reporter platform for profiling of leads in GPCR drug development. Patent USMethod and compositions for detecting.

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