This study investigated BAR130, a novel dual agonist of LXRα and GPBAR1 receptors. In vitro experiments showed that BAR130 activated both receptors in a concentration-dependent manner. When administered to mice, BAR130 did not increase liver lipids or inflammation markers. It increased expression of the GPBAR1 target gene GLP-1 in the intestine but did not induce expression of lipid metabolism genes in the liver. BAR130 represents a promising approach to target metabolic disorders by activating LXRα and GPBAR1 without causing liver lipid accumulation.
A reading report for <A Secreted Slit2 Fragment Regulates Adipose Tissue Ther...星云 王
A reading report for <A Secreted Slit2 Fragment Regulates Adipose Tissue Thermogenesis and Metabolic Function
>, only for private study use, please do not use it for profit or public.
A new effector pathway links ATM kinase with the DNA damage responseCostas Demonacos
The related kinases ATM (ataxia-telangiectasia mutated) and ATR (ataxia-telangiectasia and Rad3-related) phosphorylate a limited number of downstream protein targets in response to DNA damage. Here we report a new pathway in which ATM kinase signals the DNA damage response by targeting the transcriptional cofactor Strap. ATM phosphorylates Strap at a serine residue, stabilizing nuclear Strap and facilitating formation of a stress-responsive co-activator complex. Strap activity enhances p53 acetylation, and augments the response to DNA damage. Strap remains localized in the cytoplasm in cells derived from ataxia telangiectasia individuals with defective ATM, as well as in cells expressing a Strap mutant that cannot be phosphorylated by ATM. Targeting Strap to the nucleus reinstates protein stabilization and activates the DNA damage response. These results indicate that the nuclear accumulation of Strap is a critical regulator in the damage response, and argue that this function can be assigned to ATM through the DNA damage-dependent phosphorylation of Strap.
A reading report for <A Secreted Slit2 Fragment Regulates Adipose Tissue Ther...星云 王
A reading report for <A Secreted Slit2 Fragment Regulates Adipose Tissue Thermogenesis and Metabolic Function
>, only for private study use, please do not use it for profit or public.
A new effector pathway links ATM kinase with the DNA damage responseCostas Demonacos
The related kinases ATM (ataxia-telangiectasia mutated) and ATR (ataxia-telangiectasia and Rad3-related) phosphorylate a limited number of downstream protein targets in response to DNA damage. Here we report a new pathway in which ATM kinase signals the DNA damage response by targeting the transcriptional cofactor Strap. ATM phosphorylates Strap at a serine residue, stabilizing nuclear Strap and facilitating formation of a stress-responsive co-activator complex. Strap activity enhances p53 acetylation, and augments the response to DNA damage. Strap remains localized in the cytoplasm in cells derived from ataxia telangiectasia individuals with defective ATM, as well as in cells expressing a Strap mutant that cannot be phosphorylated by ATM. Targeting Strap to the nucleus reinstates protein stabilization and activates the DNA damage response. These results indicate that the nuclear accumulation of Strap is a critical regulator in the damage response, and argue that this function can be assigned to ATM through the DNA damage-dependent phosphorylation of Strap.
Generation of MRP2 Efflux Transporter Knock-Out in HepaRG Cell Linemdmitc
MilliporeSigma's Jennifer Pratt recently presented a poster at the 2016 AAPS/ITC Transporter Workshop demonstrating the utility of HepaRG MRP2 Knockout cells for investigating drug-transporter interactions in the liver involving MRP2.
Il punto sul microbiota e le malattie umane- Covegno scientifico Attività scientifica
Il microbiota e le patologie umane- Perugia 20 aprile 2018- Convegno a Perugia-
Probiotici e trattamento delle patologie umane
Microbiota intestinale e malattie infiammatorie
Giornata di studio sul microbiota e patologie umane- Esperti a confronto sul ruolo dl microbiota nelle patologie dell' apparato digerente Perugia 20 aprile 2018
Meeting sul microbiota umano 20 aprile 2018
Focus su microbiota intestinale e malattie infiammatorie ed epatiche- Probiotici: come valutare un probiotico?
These slides describe the pathophysiology and the management of patients with liver cirrhosis and portal hypertension. The slides are at the level of post-graduate students
GPBAR1 (TGR5) regulates il 10 production from intestinal macrophages Attività scientifica
Novel therapeutic approach to intestinal inflammation by targeting GPBAR1 (TGR5) stimulates release of anti-inflammatory cytokine IL-10.
Just published in J. Immunology June 2017
Nuovo website:
www.gastroenterologia.unipg.it
disponibile da oggi per aggiornamenti, didattica, blog ed informazioni sulla sezione di gastroenterologia di Perugia
BILE ACID ACTIVATED RECEPTORS: FROM MEDICINAL CHEMISTRY TO CLINICAL APPLICATIONSAttività scientifica
A scientific meeting on bile acids and their receptors and clinical applications in liver and metabolic disorders will be held on Feb 9, 2016 in Perugia. Attendance is free.
Inf: stefano.fiorucci@unipg.it
A seminar on nanotechnology at the Department of Surgical and Biomedical Science University of Perugia. The seminar will describe general concepts and biomedical applications of nanotechnologies.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...
Digestive Disease Week 2017 Discovery of dual LXR/GPBAR1
1. BAR130, a hyodeoxycholic derivative as the first example of dual LXRα/GPBAR1 agonist
Carino Adriana*, Cipriani Sabrina #, Marchianò Silvia*, Biagioli Michele* , Zampella Angela † , De Martino Simona † and Fiorucci Stefano*
*Dipartimento di Scienze Chirurgiche e Biomediche, Università degli studi di Perugia Nuova Facoltà di Medicina e Chirurgia, Sant’Andrea delle Fratte, Perugia, Italy
#Dipartimento di Medicina, Università degli studi di Perugia Nuova Facoltà di Medicina e Chirurgia, Sant’Andrea delle Fratte, Perugia, Italy
†Dipartimento di Farmacia, Università di Napoli Federico II, Napoli, Italy
Background.The liver X receptors (LXRs), activated by oxysterol and by secondary bile acid hyodeoxycholic acid (HDCA), heve been found essential in the regulation of lipid homeostasis in mammals. Unfortunately, LXRα activates lipogenic
enzymes causing accumulation of lipid in the liver. In the present study, we report the identification of a side chain modified HDCA (BAR130) that represents the first example of a potent dual LXRα/GPBAR1 agonist.
Aim of the study. To investigate in vitro and in vivo effects of BAR130, a dual GPBAR1/LXRα ligand.
Material and Methods. HepG2 and Glutag cells (a mouse counterpart of intestinal L cells) were exposed to GW3965 or TLCA (10µM), HDCA (10µM) along with increasing concentrations of BAR130 (1, 5, 10, 25, 50 µM); total RNA was isolated for RT-PCR.
C57BL6 mice were administered with BAR130 (30 mg/Kg daily by gavage) or vehicle (distilled water) for 2 weeks. Mice were sacrificed and blood, liver and terminal ileum collected for plasma AST, cholesterol and tryglicerides measurement, liver hystopathology
analysis (H/E) and for evaluation of steatosis marker genes expression (RT-PCR).
Results. BAR130 transactivates LXRα and GPBAR1 with an EC50 value of 3.2±0.03 and 4.9±0.02 µM, respectively. When administered in vitro to HepG2 cells, BAR130 increases the expression of LXRα target genes, ABCA1 and SREBP1C, in a concentration-
dependent manner with an EC50 of 8.3 µM and 5.8 µM respectively. BAR130 increases the expression of pro-glucagon mRNA (a GPBAR1 target gene) in GLUTAG cells with an EC50 of 6.5 µM. Treating mice with BAR130 did not increased AST, cholesterol
and tryglicerides plasma levels. Further on, rt-PCR analysis of liver samples demonstrated that the BAR130 did not induce the expression of lipidogenic genes including FAS, SREBP1C and CD36. The result was confirmed by Liver histology (H/E). BAR130
increased the expression of GPBAR1 target gene GLP1 in the ileum and reduced markers of inflammation in macrophages.
Conclusions. This study describes the first dual LXRα and GPBAR1 ligand. BAR130 represents a promising approach to target metabolic disorders.
Figure3: Effects of BAR130 on hepatic lipid metabolism and on terminal ileum after administration on intact mice. C57BL6
mice were treated with BAR130 (30 mg/Kg daily per os) for two weeks. Results are the mean ± SE of 3-5 mice per group; *p<0.05
versus control mice. Serum levels of AST, total cholesterol and triglycerides; Hematoxilin and Eosin liver staining; Relative hepatic
mRNA expression of genes involved in fatty acids metabolism (FASN, SREBP1C, CD36) and genes for nuclear receptors (PPARα,
PPARγ, LXR α); Relative mRNA expression of GLP-1, FGF21, FXR and LXRα genes in terminal ileum. (Figure 3, A-C)
Figure 1: Concentration-response curves for BAR130. HepG2 cells were transiently transfected
with the reporter vector p(UAS)5XTKLuc, a vector containing the ligand binding domain of LXRα
cloned upstream of the GAL4-DNA binding domain (i.e. pSG5-LXRαLBD-GAL4DBD) and with the
reporter vector pGL4.70 (Promega), encoding the human Renilla gene. To evaluate GPBAR1
mediated transactivation, HEK-293T cells were transfected with pGL4.29 (Promega), a reporter vector
containing a cAMP response element (CRE) that drives the transcription of the luciferase reporter
gene luc2P, with pCMVSPORT6-human GPBAR1, and with pGL4.70 Renilla. 24 hours post-
transfection cells were stimulated with increasing concentrations of BAR130 (1, 5, 10, 25 and 50 µM).
GW3965 (10 µM) or TLCA (10 μM) were used as a positive control.
For calculation of efficacy data, maximal transactivation of LRE or CRE caused by the compound (10
μM) was compared to maximal transactivation caused by GW3965 (10 µM) or TLCA (10 μM) and by
HDCA (10 μM). (Figure 1, A-C).
Figure 2: In vitro pharmacological evaluation of BAR130 activity.
Quantitative Real-Time PCR analysis of mRNA expression on LXRα
and GPBAR1 target genes. ABCA1 and SREBP1c expression in
HepG2 cells primed with increasing concentration of BAR130 (1, 5, 10
and 25 μM). GW3965 and HDCA were used as positive controls. Pro-
glucagon expression in Glutag cells stimulated with increasing dose of
BAR130 (1, 10, 25 and 50 μM). TLCA and HDCA were used as a
positive control. As shown BAR130 was able to induce the expression
of ABCA1 and SREBP1c genes in HepG2 cells in dose-dependent
manner with an EC50 of 8.3 µM and 5.8 µM respectively, and the
expression of pro-glucagon mRNA in Glutag cells; however, the
induction is dose-dependent only until the 10 µM concentration with an
EC50 of 6.5 µM. Values are normalized to GAPDH and are expressed
relative to those of not treated cells (NT) which are arbitrarily settled to
1. The relative mRNA expression is expressed as 2(-ΔΔCt). *p < 0.05
vs NT (Figure 2, A-C).
0
25
50
75
100
125
10-7 10-6 10-5 10-4
EC50 :3.2 0.03 M
0
130 (M)
%MaximalResponse
0
25
50
75
100
125
10-7 10-6 10-5 10-4
EC50: 4.990.2 M
0
130 (M)
%MaximalResponse
Compound GPBAR1* LXRa**
Efficacy Efficacy
(% vs TLCA) (% vs HDCA) (% vs GW3965) (% vs HDCA)
HDCA 26 15,5
BAR130 55,1 211,9 109,3 703,3
HepG2
NT GWHDCA 1 5 10 25
0
10
20
30
130
(M)
*
*
*
*
EC50:8.3M
ABCA1mRNArel.expression
A. B.
C.
HepG2
NT GW HDCA 1 5 10 25
0
5
10
15
20
25
130
(M)
*
* *
*
*
* EC50 : 5.8 M
SREBP1cmRNArel.expression
GLUTAG
NT TLCA HDCA 1 10 25 50
0
1
2
3
4
130
(M)
*
*
*
*
EC50 : 6.5 M
Pro-Glucagon
mRNArel.expression
A.
B.
C.
CTRL BAR130
0
50
100
150
200
ASTU/L
CTRL BAR130
0
20
40
60
80
100
TotalCholesterolmg/dL
CTRL BAR130
0
50
100
150
Triglyceridesmg/dL
0
1
2
3
FASNrelativemRNAexpression
(x10^-1)
0.0
0.5
1.0
1.5
2.0
SREBP1crelativemRNAexpression
(x10^-1)
0
1
2
3
4
5
CD36relativemRNAexpression
(x10^-3)
0
1
2
3
4
5
PPARrelativemRNAexpression
(x10^-2)
0
1
2
3
4
5
PPARrelativemRNAexpression
(x10^-3)
0
1
2
3
4
LXRrelativemRNAexpression
(x10^-2)
0
2
4
6
8
10
*
GLP-1relativemRNAexpression
(x10^-2)
0
5
10
15
FGF21relativemRNAexpression
(x10^-5)
0.0
0.5
1.0
1.5
2.0
FXRrelativemRNAexpression
(x10^-2)
0.0
0.5
1.0
1.5
2.0
LXRrelativemRNAexpression
(x10^-2)
Liver histopathology (H&E) 10x
BAR130 (30 mg/kg)Control
A.
B.
C.