1) The study found that red wines from France increased expression of the endothelial nitric oxide synthase (eNOS) gene and protein in human endothelial cells, whereas red wines from Germany did not.
2) French red wines were also found to increase activity of the eNOS promoter and stability of eNOS mRNA, resulting in increased production of bioactive nitric oxide (NO) from the endothelial cells.
3) The increase in eNOS expression, activity, and NO production brought about by French red wines may contribute to the cardiovascular benefits associated with moderate red wine consumption.
Accessing genetically tagged heterocycle libraries via a chemoresistant DNA s...Laura Berry
Presented at the Global Medicinal Chemistry and GPCR Summit. To find out more, visit:
www.global-engage.com
Andreas Brunschweiger, an Independent Group Leader at TU Dortmund, discusses the limitations of DNA-encoded compound libraries (DELs) and getting around these.
The microbiology of the winemaking process, which includes inoculated strains
of the yeast Saccharomyces cerevisiae and the lactic acid bacterium, Oenococcus
oeni, is critical to process efficiency and wine quality. In each case these organisms
are required to complete a core conversion (sugar to ethanol or lactate to malate,
respectively) as well as make desirable sensory contributions. These activities
typically occur under extreme conditions which may include high sugar (osmolarity)
and ethanol content and low pH, temperature and nutrient availability. We have used
mutant screening strategies and functional genomic approaches to identify the basis
of superior yeast performance in the face of these challenges. In addition we have
use adaptive evolution to yield yeast with enhance fermentation reliability based on
increase nitrogen efficiency, fructophilicity or general robustness. In parallel work,
we have isolated and heterologously expressed genes from O. oeni which encode
esterases or glucosidases. Characterisation of these gene products has provided
insights into their roles within the cell as well as potential contribution to wine.
USMLE Step 1 Molecular Biology and Biochemistry reviewAbril Santos
This is a review for the USMLE Step 1 exam for Molecular Biology and Biochemistry.
It contains anything you need to know for your exam in pictures, tables and diagrams.
Pescando genes: Peces que nos enseñan a entender enfermedades por Agustín G. Zapata (Departamento de Biología Celular-Universidad Complutense de Madrid) en la
Escuela de Verano "Ortega y Gasset" de la Universidad Internacional Menendez Pelayo. UIMP 2008. Palacio de la Magdalena, Santander
Accessing genetically tagged heterocycle libraries via a chemoresistant DNA s...Laura Berry
Presented at the Global Medicinal Chemistry and GPCR Summit. To find out more, visit:
www.global-engage.com
Andreas Brunschweiger, an Independent Group Leader at TU Dortmund, discusses the limitations of DNA-encoded compound libraries (DELs) and getting around these.
The microbiology of the winemaking process, which includes inoculated strains
of the yeast Saccharomyces cerevisiae and the lactic acid bacterium, Oenococcus
oeni, is critical to process efficiency and wine quality. In each case these organisms
are required to complete a core conversion (sugar to ethanol or lactate to malate,
respectively) as well as make desirable sensory contributions. These activities
typically occur under extreme conditions which may include high sugar (osmolarity)
and ethanol content and low pH, temperature and nutrient availability. We have used
mutant screening strategies and functional genomic approaches to identify the basis
of superior yeast performance in the face of these challenges. In addition we have
use adaptive evolution to yield yeast with enhance fermentation reliability based on
increase nitrogen efficiency, fructophilicity or general robustness. In parallel work,
we have isolated and heterologously expressed genes from O. oeni which encode
esterases or glucosidases. Characterisation of these gene products has provided
insights into their roles within the cell as well as potential contribution to wine.
USMLE Step 1 Molecular Biology and Biochemistry reviewAbril Santos
This is a review for the USMLE Step 1 exam for Molecular Biology and Biochemistry.
It contains anything you need to know for your exam in pictures, tables and diagrams.
Pescando genes: Peces que nos enseñan a entender enfermedades por Agustín G. Zapata (Departamento de Biología Celular-Universidad Complutense de Madrid) en la
Escuela de Verano "Ortega y Gasset" de la Universidad Internacional Menendez Pelayo. UIMP 2008. Palacio de la Magdalena, Santander
DNA damage repair Neil3 gene Knockout in MOLT-4iosrjce
RNAi is superannuated cellular mechanism that protect organism against viruses that replicate
through double- stranded RNA. RNAi can be used to diminish gene expression from plasmid expressing and
inserted sequence repeat. A stable harpin would be expressed after the vector was integrated into the genome.
In this paper a shiRNA expressing vector for Neil3 was designed and developed which is capable of replication
in MOLT-4. This shiRNA vector had the ability to arose the RNAi pathway, and reduce the gene expression of
Neil3. This was assessed by using pSilence 4.1CMV as a vector, and Gapdh as positive control.
This analysis deals with finding the residues of Endosulfan in blood. Researchers, Atmakuru Ramesh and Perumal Elumalai Ravi tested the blood samples of workers and people exposed to Endosulfan for a long time. The study concludes the absence of endosulfan residues in the blood reports.
Strategie nutraceutiche per ridurre l'infiammazione.CreAgri Europe
I polifenoli estratti dalle olive sono in grado di ridurre l'infiammazione mediata da TNF alfa. Esiste inoltre un effetto sinergico nella riduzione dello stato infiammatorio tra idrossitirosolo e glucosammina.
- inglese - testo scientifico -
Stable 16 year storage of DNA purified with the QIAamp® DNA Blood mini kit - ...QIAGEN
In this application note, we describe the success of the QIAamp DNA Blood Mini Kit in the preparation of highly stable DNA,as evidenced by 16-year storage data. We also report the best storage conditions for maximal protection against degradation.
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
NVBDCP.pptx Nation vector borne disease control programSapna Thakur
NVBDCP was launched in 2003-2004 . Vector-Borne Disease: Disease that results from an infection transmitted to humans and other animals by blood-feeding arthropods, such as mosquitoes, ticks, and fleas. Examples of vector-borne diseases include Dengue fever, West Nile Virus, Lyme disease, and malaria.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
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Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
These lecture slides, by Dr Sidra Arshad, offer a quick overview of the physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar lead (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
6. Describe the flow of current around the heart during the cardiac cycle
7. Discuss the placement and polarity of the leads of electrocardiograph
8. Describe the normal electrocardiograms recorded from the limb leads and explain the physiological basis of the different records that are obtained
9. Define mean electrical vector (axis) of the heart and give the normal range
10. Define the mean QRS vector
11. Describe the axes of leads (hexagonal reference system)
12. Comprehend the vectorial analysis of the normal ECG
13. Determine the mean electrical axis of the ventricular QRS and appreciate the mean axis deviation
14. Explain the concepts of current of injury, J point, and their significance
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. Chapter 3, Cardiology Explained, https://www.ncbi.nlm.nih.gov/books/NBK2214/
7. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
The Gram stain is a fundamental technique in microbiology used to classify bacteria based on their cell wall structure. It provides a quick and simple method to distinguish between Gram-positive and Gram-negative bacteria, which have different susceptibilities to antibiotics
Best Ayurvedic medicine for Gas and IndigestionSwastikAyurveda
Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
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2. Red wine increases the expression of human
endothelial NO synthase (eNOS):
a mechanism that may contribute to its
beneficial cardiovascular effects
Department of Pharmacology, Johannes Gutenberg University,
D-55101 Mainz, Germany
Thomas Wallerath, PhD Daniela Poleo
Huige Li MD, PhD Ulrich Förstermann, MD, PhD
3. Abstract
Objectives: In the present study, we tested the effect of red wine on
eNOS expression and eNOS activity in human endothelial cells.
Background: Endothelial-type NO synthase (eNOS) exerts
vasoprotective effects. Moderate alcohol consumption has been
associated with a reduction of cardiovascular disease, and red wine
seems to offer more benefits than any other kind of drink. However,
the molecular basis of this protective effect is unclear.
Methods: Human endothelial cells were treated with red wine and
eNOS mRNA expression was measured by RNase protection assay,
eNOS protein expression by Western blotting, and eNOS activity by
RFL-6 reporter cell assay. eNOS promoter activity was analyzed in
transfected endothelial cells; binding activities of relevant
transcription factors were determined by electrophoretic mobility
shift assay.
Results: Incubation of endothelial cells with red wines from France
upregulated eNOS mRNA and protein expression. In contrast, red
wines from Germany showed little or no effect on eNOS expression.
No significant difference in eNOS mRNA expression could be
detected between ”en barrique” (matured in oak barrels) and ”non
barrique” (matured in steel tanks) produced French red wines.
Endothelial cells treated with French red wines produced up to three
times more bioactive NO than control cells. French red wines
increased the activity of the eNOS promoter, with the essential trans-
stimulated sequence being located in the proximal 326 bp of the
promoter sequence. eNOS mRNA stability was also increased by red
wine.
Conclusions: The increase in eNOS expression and activity brought
about by red wines from France (and probably other locations) may
contribute to the beneficial effects of this beverage on the
cardiovascular system.
(J Am Coll Cardiol 2003; 41: 471-478)
4. Aggregating
platelets
Endothelial cells
Smooth muscle cells GTP cGMP
Vasodilatation
Ca2+
CaML-Arginine
+O2 L-Citrulline
R R
Ca2+
ADP
Serotonin
NO
NO
Receptor
agonists
PDGF
FGF
Proliferation
Leukocytes
Blood vessel
CD11/CD18
expression
+
+ -
-
-
NO is produced by vascular endothelium under basal conditions, and its
production can be further stimulated by a variety of receptor agonists. NO
dilates all types of blood vessels by stimulating soluble guanylyl cyclase
(sGC) and increasing cyclic cGMP in smooth muscle cells. In addition to its
vasodilator properties, NO can convey vasoprotection in several ways. NO
released towards the vascular lumen is a potent inhibitor of platelet
aggregation and adhesion to the vascular wall. Besides protection from
thrombosis, this also prevents the release of platelet-derived growth factor
(PDGF) and fibroblast growth factor (FGF) that stimulate smooth muscle
proliferation and its production of matrix molecules. Endothelial NO also
controls the expression of genes involved in atherogenesis. NO decreases
the expression of the chemoattractant protein MCP-1, and of surface
adhesion molecules such as CD11/CD18, P-selectin, vascular cell adhesion
molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) thereby
preventing leukocyte adhesion to vascular endothelium and leukocyte
migration into the vascular wall. This offers protection against an early phase
of atherogenesis. Furthermore, NO has been shown to inhibit DNA synthesis,
mitogenesis, and proliferation of vascular smooth muscle cells as well as
smooth muscle cell migration, thereby protecting against a later phase of
atherogenesis. Based on the combination of those effects, NO produced in
endothelial cells can be considered an anti-atherosclerotic principle.
Vasoprotective role of NO
eNOS
sGC
5. 0
50
100
150
200
250
300
350
400
450
500
eNOSmRNAlevel(%ofcontrol)
***
***
***
***
***
***
German red wines
Bordeaux
Burgundy
Rhone French red wines
Effect of red wines on eNOS mRNA expression in human endothelial
EA.hy 926 cells. Cells were left untreated (control, Co) or exposed for 24 h
to ethanol (EtOH, 1.25%, v/v), three different German red wines
("Dornfelder 1997“ (1), "Kriegsheimer Rosengarten 1997“ (2) and
"Spätburgunder 1999“ (3)) or six different French red wines from
Bordeaux ("Chateau Bonnet reserve 1997“ (4), "Chateau Bellegrave 1997“
(5) and "Chateau de la Marechaude 1997“ (6)), Rhone ("Chateau Cabieres
1998“ (7)), and Burgundy ("Chateau Rousseau 1998“ (8), "Les Chevaliers
de la Reine 1998“ (9)). In this experiment, the red wines were diluted in
the cell culture medium to a final concentration of 10% (v/v). RNase
protection analyses were performed using antisense RNA probes to
human eNOS and GAPDH (for standardization).
Co EtOH 1 2 3 4 5 6 7 8 9
Effect of different red wines on eNOS mRNA
expression in human endothelial cells
6. Effect of French red wines on eNOS mRNA
expression in human endothelial cells
eNOS
GAPDH
EtOH MT N GLes Chev.
10%
EtOH
***
Les Chev.
10%
0
50
100
150
200
250
300
350
400
450
eNOSmRNAlevel(%of
control)
118
140
151
200
249
311
417
100
nt
Upregulation of eNOS mRNA by the French red wine "Les Chevaliers de la
Reine 1998" in primary human umbilical vein endothelial cells (HUVEC).
eNOS mRNA expression was quantified with RNase protection assay. RNA
was prepared from HUVEC either incubated for 24 h with ethanol (EtOH,
1.25%, v/v) or with the red wine "Les Chevaliers de la Reine 1998" (Les Chev.,
10% v/v).
0
50
100
150
200
250
300
EtOH Chateau Bonnet
`non-barrique`
**
***
Chateau Bonnet
`en -barrique`
eNOSmRNAlevel(%ofcontrol)
n. s.
Upregulation of eNOS mRNA in human endothelial cells by the French red
wines "Chateau Bonnet 1997“ (“non-barrique“, matured in steel tanks) and
"Chateau Bonnet reserve 1997“ (“en-barrique“, matured in oak barrels).
eNOS mRNA expression was quantified with RNase protection assay. RNAs
were prepared from EA.hy 926 cells either incubated for 24 h with ethanol
(EtOH, 1.25%, v/v) or with the red wines (10% v/v).
7. B
100
120
140
160
180
200
0 12 24 36 48 60
Incubation time (h)
***
***
***
***
** **
*
*
10%
3%
1%
Ch.Bonnet
10 d
**
*
eNOSmRNAlevel(%ofcontrol)
A
**
***
***
Dornfelder Ch.Bonnet L.Chevaliers
1% 3% 10% 1% 1%3% 3%10% 10%EtOH
100
150
200
250
0
Concentration- and time-dependent upregulation of eNOS mRNA expression
by French red wines. eNOS mRNA expression was quantified with RNase
protection assay. Panel A: EA.hy 926 cells were incubated for 24 h with
ethanol (EtOH, 1.25%, v/v,), a German red wine ("Dornfelder 1997") and two
French red wines ("Les Chevaliers de la Reine 1998" and "Chateau Bonnet
reserve 1997") diluted to final concentrations of 1%, 3% and 10% (v/v). Panel
B: Cells were exposed to cell culture medium containing 1%, 3% and 10%
(v/v) of " Chateau Bonnet reserve 1997" for different periods of time.
Effect of different red wines on eNOS mRNA
expression in human endothelial cells
eNOSmRNAlevel(%ofcontrol)
8. Effect of French red wines on eNOS protein expression
and NO production in human endothelial cells
eNOS
Les Chevaliers de
la Reine
β-Tub.
Chateau Bonnet
EtOHEtOH 1% 1% 3%3% 10% 10%
A
NOproduction(%ofcontrol)
0
50
100
150
200
250
300
350
400
450
Chateau Bonnet
EtOH 10%1%
Les Chevaliers
de la Reine
3% 1% 3% 10%
B
**
*
*
**
Effect of French red wines on eNOS protein expression and eNOS activity in
endothelial EA.hy 926 cells. Panel A: Western blot analyses using a
polyclonal anti-eNOS- and a monoclonal anti-β−tubulin-antibody (for
normalization). Cells were incubated for 48 h with ethanol (EtOH, 1.25 %,
v/v) or two different French red wines ("Chateau Bonnet reserve 1997" and
"Les Chevaliers de la Reine 1998", 1%, 3% and 10%, v/v). The blots
represent four independent experiments each. Panel B shows the effect of a
48 h incubation with "L.Chevaliers" and "Ch.Bonnet reserve" on the NO
production in EA.hy 926 cells stimulated for 2 min with the calcium
ionophore A23187. Conditioned medium from EA.hy 926 cells was
transferred to RFL-6 reporter cells and incubated for 2 min. Thereafter,
cGMP content in RFL-6 cells was determined by radioimmunoassay.
9. Effect of French red wines on eNOS promoter
activity in human endothelial cells
eNOSpromoteractivity
(%ofcontrol)
0
50
100
150
200
250
300
350
400
**
***
Chateau Bonnet Les Chevaliers
de la Reine
EtOH 1% 3% 10%
***
***
1% 3% 10%
A
0 100 200 300
eNOS promotor activity (% of each control)
pGl3-
basic
+1
-1600
Luc
+1
-1111
Luc
+1
-633
Luc
+1
-326
Luc
B
150 25050 350
EtOH
Les Chevaliers 10%
Enhancement of eNOS promoter activity in EA.hy 926 cells after incubation
with French red wines. Panel A: EA.hy 926 cells were stably transfected with
p-eNOS-3500-Hu-Luc-neo. Stable cells were incubated for 24 h with ethanol
(EtOH, 1.25%, v/v), or various concentrations of two French red wines ("Les
Chevaliersde la Reine 1998" and "Chateau Bonnet reserve 1997"). The
luciferase activity (normalized by protein content) was taken as a measure of
eNOS promoter activity. Panel B: EA.hy 926 cells were transiently transfected
with pGl3-Basic (containing a promoter-less luciferase gene) or different eNOS
promoter luciferase constructs (p-eNOS-1600-Hu-Luc, p-eNOS-1111-Hu-Luc, p-
eNOS-633-Hu-Luc and p-eNOS-326-Hu-Luc, containing 1.6-kb to 0.33-kb of the
eNOS promoter cloned before a luciferase reporter gene). The different
transfected cells were either exposed 24 h to ethanol (EtOH, 1.25%, v/v) or to
the red wine "Les Chevaliers de la Reine 1998" (10%, v/v). The relative
luciferase activity (corrected with renilla-luciferase activity) was taken as a
measure of eNOS promoter activity.
10. Effect of a French red wines on eNOS mRNA
stability
eNOSmRNAlevel(%ofcontrol)
***
**
***
0 12 24 36
Time (h) after actinomycin D treatment
EtOH
Ch.Bonnet 10%
0
40
60
80
100
120
20
Effect of French red wine on eNOS mRNA stability. EA.hy 926 cells were pre-
incubated with ethanol (EtOH, 1.25%, v/v) or with or the red wine "Chateau
Bonnet reserve 1997" (10%, v/v) for 24 h. To inhibit transcription, actinomycin
D (20 µg/mL) was added to the culture medium. RNA was prepared 0, 12, 24
and 36 h thereafter. The eNOS mRNA expression was determined by RNase
protection assay. The eNOS mRNA levels in both groups at the time of
addition of actinomycin D (0 h) were set at 100%.
Red wine
eNOS mRNA
expression
eNOS protein
expression
eNOS promoter
activity
eNOS mRNA
stability
bioactive NO
Summary:
vasoprotection ?!