The document discusses the role of salivary gland epithelial cells in the pathogenesis of Sjögren's syndrome. It finds that in Sjögren's syndrome patients, salivary gland epithelial cells lose their polarity and detach from the basal lamina. This is due to increased MMP expression degrading extracellular matrix proteins. While patients also show remodeling of the basal lamina and new cell-cell interactions to maintain organization. The loss of polarity results in aberrant exocytosis of mucins into the extracellular matrix. Exogenous mucins induce the expression of pro-inflammatory cytokines in cultured salivary cells. Together this suggests salivary gland epithelial cells actively participate in Sjögren's syndrome pathogenesis through the loss of polarity
Genetic Insights Into Multiple Sclerosis PathogenesisAaron Sparshott
A segment of a group presentation reflecting upon some of the genetic components that may contribute to Multiple Sclerosis pathogenesis.
IL2Rα and IL7Rα were the two genes of focus.
(This presentation was originally done for Semester 2 , 2008)
Genetic Insights Into Multiple Sclerosis PathogenesisAaron Sparshott
A segment of a group presentation reflecting upon some of the genetic components that may contribute to Multiple Sclerosis pathogenesis.
IL2Rα and IL7Rα were the two genes of focus.
(This presentation was originally done for Semester 2 , 2008)
Patients with systemic autoimmune diseases such as lupus and rheumatoid arthritis have a striking increase in risk to develop vascular complications such as myocardial infarction and stroke. Immune dysregulation characteristic of these diseases appears to play a prominent role in this enhanced risk.
In this presentation, Dr. Mariana Kaplan highlights the role of the immune system in the development of vasculopathy and premature atherosclerosis in systemic autoimmunity and discuss potential strategies to prevent these devastating complications.
Key Topics Include:
- To review the epidemiology of premature vascular disease in systemic autoimmunity
- To review the pathogenic mechanisms that lead to vascular damage in patients with autoimmunity
- To discuss potential therapeutic strategies to prevent/treat vascular damage in patients with autoimmune diseases
Identification of genetic regions in the yuk operon of Bacillus subtilis that are differentially required for secretion of YukE, a homolog to the virulence factor, ESXA in Mycobacterium tuberculosis
Gene therapy is that the introduction of a traditional gene into an individual’s genome so as to repair a mutation that causes a genetic disorder.
When a traditional gene is inserted into a mutant nucleus, it presumably will integrate into a chromosomal site different from the defective allele; although this might repair the mutation, a replacement mutation may result if the traditional gene integrates into another functional gene.
Human gene therapy has been attempted only on somatic (body) cells for diseases like cancer and severe combined immunodeficiency syndrome (SCIDS).
Patients with systemic autoimmune diseases such as lupus and rheumatoid arthritis have a striking increase in risk to develop vascular complications such as myocardial infarction and stroke. Immune dysregulation characteristic of these diseases appears to play a prominent role in this enhanced risk.
In this presentation, Dr. Mariana Kaplan highlights the role of the immune system in the development of vasculopathy and premature atherosclerosis in systemic autoimmunity and discuss potential strategies to prevent these devastating complications.
Key Topics Include:
- To review the epidemiology of premature vascular disease in systemic autoimmunity
- To review the pathogenic mechanisms that lead to vascular damage in patients with autoimmunity
- To discuss potential therapeutic strategies to prevent/treat vascular damage in patients with autoimmune diseases
Identification of genetic regions in the yuk operon of Bacillus subtilis that are differentially required for secretion of YukE, a homolog to the virulence factor, ESXA in Mycobacterium tuberculosis
Gene therapy is that the introduction of a traditional gene into an individual’s genome so as to repair a mutation that causes a genetic disorder.
When a traditional gene is inserted into a mutant nucleus, it presumably will integrate into a chromosomal site different from the defective allele; although this might repair the mutation, a replacement mutation may result if the traditional gene integrates into another functional gene.
Human gene therapy has been attempted only on somatic (body) cells for diseases like cancer and severe combined immunodeficiency syndrome (SCIDS).
Dr. masciotra shear wave elastography and more in a clinical case of multip...antonio pio masciotra
This is the presentation of a clinical case of Multiple Myeloma with a thorough discussion on many aspects of the disease and on applications of Shear Wave Elastography.
Anti-Mullerian hormone (AMH) is a glycoprotein, a member of the transforming growth factor-B super family. This hormone is a sensitive marker of ovarian reserve. The present study aims to measure the Anti-Mullerian hormone in thalassemic females receiving the regular blood transfusion as well as patients of chronic idiopathic thrombocgtopenic purpura and age and sex matched controls. Serum Anti-Mullerian hormone was measured by ELISA and Ferritin were measured by RIA. Clinical evaluation was done for all patients including anthropometric measurements, pubertal staging and history taking. Results of the study were analyzed by appropriate statistical methods. Obtained results revealed that the values of Body Mass Index as well as Anti-Mullerian were significantly higher in controls than thalassemics and chronic idiopathic thrombocytopenic purpura and there was a negative correlation between serum Ferritin and Anti-Mullerian hormone. Moreover, Anti-Mullerian hormone was significantly higher in patients receiving Desferal than in those receiving Deferriprone. Reduced Anti-Mullerian hormone in thalassemics as well as chronic ITP patients are considered an important indicator declines in ovarian function which entail modification in the therapeutic plans for thalassemic and chronic ITP patients.
Biomarkers of Systemic Lupus Erythematosus and Systemic Sclerosis diseases ac...Prof. Hesham N. Mustafa
Systemic Lupus Erythematosus (SLE) and systemic sclerosis (SSc) are systemic inflammatory autoimmune disorders characterized by a large spectrum of clinical and laboratory features. The aim of the present study was to investigate the possible use of serum level of soluble intercellular adhesion molecule-1(sICAM-1) and soluble interleukin-2 receptor (sIL-2Ra) as biomarkers for monitoring of SLE and SSc disease activity. Moreover, it aimed to compare the specificity and sensitivity as well as cut-off value of both biomarkers in a sample of Egyptian patients. 50 SLE patients, 30 SSc patients and 60 age and sex matched healthy controls were enrolled in our study. sICAM-1and sIL-2Ra were measured in serum samples obtained from all participants. In addition to Erythosedimentation rate (ESR), complete blood count (CBC), Antineuclearantibodies (ANA) estimation, disease activity of both diseases were also assessed. sICAM-1and sIL-2Ra levels were higher in SLE and SSc patients versus control. Both parameters are correlated with each other as well as the activity parameters. A cut-off levels of 455.59 (ng/ml) &2525935 (pg/ml) in both SLE & SSs respectively was observed with the highest specificity and sensitivity. It could be concluded that sICAM-1 and sIL-2Ra are noninvasive biomarkers for SLE and SSc that could play a pathophysiologic role in development and progression of both diseases. Moreover, sICAM-1 and sIL-2Ra are correlated with the disease activity at cut-off values of 455.59 (ng/ml) & 2525935(pg/ml) respectively.
Experimental Analysis of Tear Fluid and their Processing for the Diagnosis of...semualkaira
A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using various experimental methods: liquid chromatography-mass spectrometry, Raman spectroscopy, infrared spectroscopy, and atomic force microscopy. Infrared spectroscopy found no significant difference in both MS patients and control spectra of tear fluid; all three significant peaks were located at around the same position
Experimental Analysis of Tear Fluid and their Processing for the Diagnosis of...semualkaira
A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using
various experimental methods: liquid chromatography-mass spectrometry, Raman spectroscopy
Experimental Analysis of Tear Fluid and their Processing for the Diagnosis of...semualkaira
A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using
various experimental methods: liquid chromatography-mass spectrometry, Raman spectroscopy, infrared spectroscopy, and atomic
force microscopy. Infrared spectroscopy found no significant difference in both MS patients and control spectra of tear fluid; all
three significant peaks were located at around the same position
Experimental Analysis of Tear Fluid and their Processing for the Diagnosis of...semualkaira
A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using
various experimental methods: liquid chromatography-mass spectrometry, Raman spectroscopy, infrared spectroscopy, and atomic
force microscopy. Infrared spectroscopy found no significant difference in both MS patients and control spectra of tear fluid; all
three significant peaks were located at around the same position.
Raman analysis showed the differences between the spectrum of
tear fluid of the MS patients and the spectrum of healthy subjects,
which indicate a decrease in tryptophan and phenylalanine content
and changes in the relative contributions of secondary structures
of polypeptide chains of tear proteins
Experimental Analysis of Tear Fluid and their Processing for the Diagnosis of...semualkaira
A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using
various experimental methods: liquid chromatography-mass spectrometry, Raman spectroscopy, infrared spectroscopy, and atomic
force microscopy.
Experimental Analysis of Tear Fluid and their Processing for the Diagnosis of...semualkaira
A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using
various experimental methods: liquid chromatography-mass spectrometry, Raman spectroscopy, infrared spectroscopy, and atomic
force microscopy. Infrared spectroscopy found no significant difference in both MS patients and control spectra of tear fluid
Experimental Analysis of Tear Fluid and their Processing for the Diagnosis of...semualkaira
A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using
various experimental methods: liquid chromatography-mass spectrometry, Raman spectroscopy, infrared spectroscopy, and atomic
force microscopy. Infrared spectroscopy found no significant difference in both MS patients and control spectra of tear fluid
Similar to 8. Julieta Gonzalez. Bogota epithelial cells in lsg from ss patients (20)
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!
CDSCO and Phamacovigilance {Regulatory body in India}NEHA GUPTA
The Central Drugs Standard Control Organization (CDSCO) is India's national regulatory body for pharmaceuticals and medical devices. Operating under the Directorate General of Health Services, Ministry of Health & Family Welfare, Government of India, the CDSCO is responsible for approving new drugs, conducting clinical trials, setting standards for drugs, controlling the quality of imported drugs, and coordinating the activities of State Drug Control Organizations by providing expert advice.
Pharmacovigilance, on the other hand, is the science and activities related to the detection, assessment, understanding, and prevention of adverse effects or any other drug-related problems. The primary aim of pharmacovigilance is to ensure the safety and efficacy of medicines, thereby protecting public health.
In India, pharmacovigilance activities are monitored by the Pharmacovigilance Programme of India (PvPI), which works closely with CDSCO to collect, analyze, and act upon data regarding adverse drug reactions (ADRs). Together, they play a critical role in ensuring that the benefits of drugs outweigh their risks, maintaining high standards of patient safety, and promoting the rational use of medicines.
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
263778731218 Abortion Clinic /Pills In Harare ,sisternakatoto
263778731218 Abortion Clinic /Pills In Harare ,ABORTION WOMEN’S CLINIC +27730423979 IN women clinic we believe that every woman should be able to make choices in her pregnancy. Our job is to provide compassionate care, safety,affordable and confidential services. That’s why we have won the trust from all generations of women all over the world. we use non surgical method(Abortion pills) to terminate…Dr.LISA +27730423979women Clinic is committed to providing the highest quality of obstetrical and gynecological care to women of all ages. Our dedicated staff aim to treat each patient and her health concerns with compassion and respect.Our dedicated group ABORTION WOMEN’S CLINIC +27730423979 IN women clinic we believe that every woman should be able to make choices in her pregnancy. Our job is to provide compassionate care, safety,affordable and confidential services. That’s why we have won the trust from all generations of women all over the world. we use non surgical method(Abortion pills) to terminate…Dr.LISA +27730423979women Clinic is committed to providing the highest quality of obstetrical and gynecological care to women of all ages. Our dedicated staff aim to treat each patient and her health concerns with compassion and respect.Our dedicated group of receptionists, nurses, and physicians have worked together as a teamof receptionists, nurses, and physicians have worked together as a team wwww.lisywomensclinic.co.za/
New Drug Discovery and Development .....NEHA GUPTA
The "New Drug Discovery and Development" process involves the identification, design, testing, and manufacturing of novel pharmaceutical compounds with the aim of introducing new and improved treatments for various medical conditions. This comprehensive endeavor encompasses various stages, including target identification, preclinical studies, clinical trials, regulatory approval, and post-market surveillance. It involves multidisciplinary collaboration among scientists, researchers, clinicians, regulatory experts, and pharmaceutical companies to bring innovative therapies to market and address unmet medical needs.
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
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
- Video recording of this lecture in English language: https://youtu.be/kqbnxVAZs-0
- Video recording of this lecture in Arabic language: https://youtu.be/SINlygW1Mpc
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
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
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
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.
NVBDCP.pptx Nation vector borne disease control program
8. Julieta Gonzalez. Bogota epithelial cells in lsg from ss patients
1. ARE THE SALIVARY GLAND EPITHELIAL CELLS A TARGET OR AN
ACTOR IN THE SJÖGREN'S SYNDROME PATHOGENESIS?
Simposio de Autoimunidad en Reumatología:
Septiembre 9, 2016 | Bogotá, Colombia
María Julieta González Burgos
2. Salivary Glands of control subjects and Sjögren’s syndrome
patients are highly complex system
Reflection
Our physiology is a large puzzle; we know little about
each one of the different systems
3. How can we get specific answers that help us to understand highly
complex diseases such as autoimmune diseases?
So, an intriguing question is:
For me it's a mystery
Yes
No
a dream
……but step by step the answers have slowly been emerging ¡¡¡¡
4. General aspects of Sjögren's syndrome patients
Sjögren's syndrome is a chronic autoimmune disease that mainly affects
the salivary and lachrymal glands producing severe disorders of mouth and eye
dryness.
This disease has the second highest prevalence after rheumatoid arthritis
and with a ratio of 9:1 exhibiting one of the highest female-to-male ratios among
autoimmune diseases. It is diagnosed around the fourth and fifth decade and its
progress is insidious.
The symptoms are unclear and difficult to diagnose, causing much
frustration for the patient who becomes a conflictive person for their family,
friends and colleagues.
Like all autoimmune diseases, treatments are palliative and the action of
drugs is highly variable. As life expectancy has increased, the autoimmune
diseases have increased considerably and the scientists who study them are still
very scarce.
The National Institute of Health estimated that autoimmune disorders affected 24 million Americans (17 in 100).
5. ¿A que nivel nos ubicamos para hacer una observación?
Salvador Felipe Jacinto Dalí
What level do we stand to make an observation?
7. The role of intrinsic epithelial activation in the pathogenesis of Sjögren’s syndrome
Adapted of:
Sjögren’s Syndrome
Novel Insights in Pathogenic,
Clinical and Therapeutic
Aspects
Chapter | 12 page 195, 2016
8. Glandular epithelial cells are leading actors in the development and maintenance
of Sjögren’s syndrome autoimmune responses.
Salivary gland epithelial cells (SGECs) are suitably equipped to mediate the recruitment
activation and/or differentiation of T and B lymphocytes, macrophages (MΦs), and dendritic
cells (DCs), as well as the organization of infiltrates in the minor salivary glands (MSGs) of SS
patients through the production of cytokines/chemokines. Chapter | 12 page 198, 2016
10. Loss of Acinar Cell Polarity
Normal Acinus Altered Acinus
L
L
Control SS-patients
It is necessary to keep in mind
When we started studying this disease our first finding was to observe
acinar cells that had lost their cell polarity.
So the obvious question was “why did the acinar cells lose their attachment to the basal lamina?
11. The acinus function
depends on a polarized
architecture that is
based on continuous
dialogue between:
Acinar cells
Cells and Basal lamina
Acinar and myoepithelial
cells
ACINUS
14. ARTHRITIS & RHEUMATISM 43:2807–2817, 2000.
ARTHRITIS & RHEUMATISM 48: 2573–2584, 2003
ARTHRITIS & RHEUMATISM 52: 2751–2760, 2005
Ann Rheum Dis 2006;65;178-183
Salivary glands from SS-patients showed an increased
MMP-3 and MMP-9 expression, as well as MMP-9
catalytic activity. These enzymes were synthesized by
exocrine epithelial cells.
An altered balance between MMPs/TIMPs was detected
Salivary glandular extracts of SS-patients degraded purified
ECM proteins, such as laminins, fibronectin, type 1, III and
IV collagens, among others.
Basal lamina becomes disorganized as a result of
degradation of at least two components, laminin 1 and
type IV collagen.
15. Ann Rheum Dis 2009;68:991–996.
ARTHRITIS & RHEUMATISM 54: 3465–3475,2006.
ARTHRITIS & RHEUMATISM 63: 1106–1115,2011
Lateral redistribution of α6β4 integrin and the
formation of new cell–cell interactions was observed
Significant increase in BP230 protein levels and the
formation of new hemidesmosomes was found.
Significant increase in both mRNA and protein levels
of laminin α1, α4, and 2 chains showing that the
basal lamina of SG from SS-patients is undergoing
active remodeling.
Together these results showed that the salivary glands from SS-patients display a
molecular potential to disorganize their ECM thus inducing detachment of acinar cells.
However, acinar cells also have activated mechanisms for remodeling of basal lamina and
the renewal of new cell-cell and cell-basal interactions, which help maintain acinar
organization and promote cell survival.
16. Adapted of Am J Physiol Gastrointest Liver Physiol 279: G250–G254, 2000.
AF6
Rab 3B
Rab 13
PKC
7H6
Simplekin
ASIP
BAP-1
Sec 6/8
ZAK
ZONAB
Occludin
Claudins
Microfilaments of actin
Simplekin
VAP33
JAM
CAR
ZO1
ZO1
Plasma
membrane
CAR
Coxsackievirus and Adenovirus Receptor
Cell 1 Cell 2
Pole Apical
Tight Junctions are Multiproteic Complexes
17. Protein
Relative Protein Levels (Mean ± SD)
Controls Patients p
Ratio
P/C
Claudin-1 0.2 ± 0.2 0.4 ± 0.2 0.02 2.0
Claudin-3 1.5 ± 0.4 1.2 ± 0.5 0.66 0.8
Claudin-4 0.13 ± 0.1 0.34 ± 0.4 0.04 2.6
Occludin 1.1 ± 1.2
0.5 ± 0.7 0.089 0.45
a1.32 ± 0.8 0.38 1.2
b0.13 ± 0.15 0.004 0.12
ZO-1 0.5± 0.6 0.1 ± 0.1 0.002 0.2
Tight Junction Relative Protein Levels from LSG of SS
Patients
In SS patients, Claudin 1 and 4 were up-regulated, while Occludin
and ZO1 were down-regulated. However, Claudin 3 showed no
change ARTHRITIS & RHEUMATISM 62:1280–1289, 2010
18. Claudin-3 showed re-distribution from apical
to basal in Acini and Ducts from MSG of SS patients
Acini Ducts
Control
Patient
A A´ B´
C´ D´
B
C D
`L
L
LLL
L
L
L
L
L
ARTHRITIS & RHEUMATISM 62:1280–1289, 2010
19. Ultrastructure of Tight
Junctions of Acinar
Cells
TJ ultrastructure in the acinar
cells of SS patients showed an
electron-dense material
extending from the apical to
basolateral plasma membrane.
This distribution was similar to
the Claudin 3 distribution.
Cytoplasmic vesicles containing
a similar material were also
observed. Vesicles like exosomes
were observed in the lumen
L
L
B
L
A
d
d
L
Control
Patient
ARTHRITIS & RHEUMATISM 62:1280–1289, 2010
21. What factors could be participating in this
redistribution?
In other epithelia it has been reported that cytokines
may disorganize TJ. In addition, salivary glands from SS
patients present high levels of these cytokines.
So, we incubated control acini with TNF- alone and
with both TNF- and INF- and then occludin was
identified in these samples.
23. C
L
B
L
A Control
TNF- 10ng/ml
TNF- 30ng/ml
L
D
L
TNF- + IFN- 20 ng/ml
Effect of TNF- and/or IFN- on acinus
and localization of occludin
In all experimental
conditions we observed
re-distribution of
occludin
ARTHRITIS & RHEUMATISM 62:1280–1289, 2010
24. IFN- IFN- and TNF-
(Sci STKE.2006 (316):pe1 Review)
APICAL
What cytokine induced mechanisms cause TJ Disorganization?
25. STX3
VAMP8
SNAP-23
STX3
VAMP8
STX4
STX4 and
SNAP-23
VAMP8
VAMP8
STX4 and SNAP-23
VAMP8
Munc18
Rab3D
Synaptotagmin
GTP
Syntaxin 4
SNAP-23
Ca2+
Ca2+
Ca2+
NSF
α-SNAP
Mature secretory granule
Plasma membrane
1. Rab GTPases 2. SNARE proteins 3. NSF/αSNAP 4. Modulating proteins
SNARE proteins in
control acinar cells
SNARE proteins are receptors for
membrane fusion
How do changes in cell polarity affect the localization
of proteins involved in regulated exocytosis?
26. In control acini,
VAMP8 was
localized in the
apical secretory
granules and
the nuclei were
localized in the
basal region of
acinar cells.
In SS-patients,
VAMP8 was
observed all
over the
cytoplasm and
the nuclei had
lost their
polarity.
apical
C
Patient
VAMP8/nuclei
apical
D
apical
B
apical
A
Control
Re-distribution of VAMP8 and loss of nuclear polarity in
acinar cells from MSG of SS-patients
ARTHRITIS & RHEUMATISM 63:3126–3135, 2011
27. STX3, in control acini is located in the
apical cytoplasm, while in SS-patients it is
observed in all over the cytoplasm.
STX4, in control acini, is mainly localized
next to the basolateral plasma
membrane and to a lesser extent to the
apical plasma membrane. In SS-patients
STX-4 is mainly localized in the basal
plasma membrane.
SNAP-23, in control acini, is mainly
localized in the plasma membrane, while
in SS-patients, it has lower
immunoreactivity, particularly at the
apical and lateral plasma membranes.
Subcellular localization of STX3, STX4,
and SNAP-23 in SS-patients
Journal of Autoimmunity 39: 83-92, 2012
PatientControl
SNAP-23E
apical
A STX3
apical
B
F
DC
STX4
28. C
Patient
apical
BA
apical
Control Control
apical
B
D
basal
Patient
apical
Considering the re-distribution of SNARE proteins we were interested in determining if
SNARE complexes localized ectopically were formed
1. In acinar cells of SS patients there was a significant
increase in the levels of SNARE complexes for VAMP8
and STX4.
2. In controls, VAMP8 localized to the apical
cytoplasm of acinar cells, whereas in SS-patients
VAMP8 was observed all over the cytoplasm.
Formation of SNARE-complexes in vivo
3. In controls, STX4 localized in all the plasma
membrane, in SS-patients STX4 co-localized with
VAMP8, mainly in basolateral plasma membrane
Journal of Autoimmunity 39: 83-92, 2012
Control
32
100
Monomeric STX4
P25ºC
C25ºC
100
18
kDa
Monomeric VAMP-8
P100ºC
C100ºC
P25ºC
C25ºC
kDa
STX4 SNARE Complex
VAMP8 SNARE Complex
C100ºC
P100ºC
Patient
SNARE COMPLEXES ARE STABLE AT 25ºC
29. Could these ectopic SNARE complexes participate in
the basal exocytosis of mucins?
30. Patient
Control
D
m
S
ECM
S
m
S
A
MUC7/LAMININ
B
DIC
C
MUC7/LAMININ/DIC
E F
Presence of secreted MUC7 in the extracellular matrix of MSG from SS-patients
In controls, MUC7 localized in the cytoplasm of acinar cells. In SS patients MUC7 maintained
its localization in the acinar cells, but was also detected atypically in the ECM. The Laminin a
component of the basal lamina marked the boundary of the acinus.
Journal of Autoimmunity 39: 83-92, 2012
32. So, we wanted to determine whether
exogenous mucins were able to induce the
expression of pro-inflammatory cytokines in
cultured human salivary cells (HSG).
These results allowed us to conclude that the loss of
acinar cell polarity leads to ectopic exocytosis, which
could explain the oral dryness in these patients.
34. Relative mRNA levels of pro-inflammatory cytokines
in HSG cells stimulated with mucins
A significant increase (*) in relative mRNA levels of pro-inflammatory cytokines was
observed. Differences found for BAFF were not significant.
……..So we wondered………..
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
CXCL-8 TNF-α IFN-α IFN-β IL-6 IL-1β BAFF
Cytokine/h18S(Relativeexpressionratio) 0µM
0.001 µM
0.01 µM
0.1 µM
0.2µM
0.4 µM
*
*
*
*
*
*
*
**
*
BSM
Barrera et al, under revision
35. Are the mucin’s sugar residues involved in
the induction of these cytokines?
36. Effect of purified MUC5B and Sulfo-Lewis residues in the relative
expression levels of cytokines in HSG cells
Sulfo-Lewis showed a comparable response to purified MUC5B. These results
allowed us to postulate that the mucins might be recognized by their sugar
residues.
Sulfo Lewis : SO3-Galβ1-3GlcNAc is a sulfated disaccharide presents in MUC5B
MUC5B Sulfo-Le BSM
*
*
*
*
*
*
*
*
*
*
*
0
0,5
1
1,5
2
2,5
3
3,5
4
4,5
5
0 0.005 0.015 21.6 54 0.4 µM
Cytokine/h18S(Relativeexpressionratio)
IL-6
TNF-α
IL-1
Barrera et al, manuscript under review
37. Ligands containing sugar residues can bind Toll like
receptors, such as TLR4. We used the
lipopolysaccharide (LPS), a ligand of TLR4, to induce
expression of cytokines. Then, we compared the LPS-
induced cytokine profile with the mucin-induced
profile.
38. Comparison between relative mRNA levels of pro-inflammatory
cytokines in HSG cells stimulated with LPS or mucins
Stimulation of HSG cells with TLR4 ligand induced the expression of IL-6 and TNF-α
in a similar manner to stimulation with mucins.
LPS BSM
* *
*
*
*
*
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
0 0.01 0.1 0.2 0.4 µM
Cytokine/h18S(Relativeexpressionratio)
IL-6
TNF-α
Barrera et al, manuscript under review
39. The role of TLR4 in cytokine induction was evaluated using TBX2, a peptide that
specifically inhibits TIRAP binding to TLR4. The TNF- and IL-6 expression were
determined.
TBX2: GKMADWFRQTLLKKPKKRPNSPESTLQLRDATPGGAIVS
TBX2
Modified of Cytokine 42 (2008) 145–151Modified of JBiotechnology 129 (2007) 555–564
Dominant-negative domain
TIRAPTBX2
TLR4
MyD88
Molecular modeling and prediction of protein–
protein interaction. TLR4, TIRAP, MyD88 and TBX2
are shown in ribbon representation.
40. Relative mRNA levels of TNF-α and IL-6 in HSG cells treated with a
TIRAP inhibitory peptide of TLR4
TBX2 selectively inhibited the production of pro-inflammatory cytokines under
the induction of mucins or LPS. This inhibition might have been mediated by
the disruption of the recruitment of downstream effector molecules. Mucins
induced the expression of pro-inflammatory cytokines via TLR4.
TNF-α
#
*
#
*
0
1
2
3
4
5
6
UT TBX2 LPS TBX2/LPS BSM TBX2/BSM
TNF-α/h18S(Relativeexpressionratio)
IL-6
#
*
#
*
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
UT TBX2 LPS TBX2/LPS BSM TBX2/BSM
IL-6/h18S(Relativeexpressionratio)
Barrera et al, manuscript under review
41. anti-hTLR4-IgG
Modified of Cytokine 42 (2008) 145–151
To confirm the involvement of TLR4, we used blocking antibodies
against TLR4 and measured TNF- and IL-6 expression in LPS-treated
or mucin-treated cells.
42. Relative IL-6 mRNA levels in HSG cells treated with
a TLR4-blocking antibody
The specific antibody against TLR4 inhibited the expression of cytokines. These
results demonstrated that mucins induced the expression of pro-inflammatory
cytokines via a TLR4 mediated mechanism.
*
*
#*
*
#
0
0.5
1
1.5
2
2.5
3
3.5
4
IL-6/h18S(Relativeexpressionratio)
Barrera et al, manuscript under review
43. In HSG cells, exogenous MUC5B and Sulfo-Lewis can induce,
via TLR4, the mRNA expression of pro-inflammatory
cytokines. However, we aren’t discarding the possibility that
galectins and TIM family receptors might also play a role.
A similar response might be observed in the salivary gland
extracellular matrix of SS patients, where the ectopically
secreted mucins may promote the infiltration of inflammatory
cells, a hallmark in the pathogenesis of the disease.
CONCLUSIONS
44. Concluding Remarks
Cellular interactions are essential for the maintenance of morphological
and functional homeostasis of salivary epithelia.
An important element involved in the development of autoimmunity is the
loss of the protective function of mucosal barriers by disruption of tight junctions.
The potentially pathogenic ectopic secretion of mucins and other factors
into the ECM “may be the initial step in the genesis of the disease”.
45. Colaborators
National
Sergio Aguilera
Claudio Molina
Sergio González
Cecilia Alliende
Marcela Hermoso
Ulises Urzúa
Andrew Quest
Lisette Leyton
Cecilia Leyton
Internacional
Juan Manuel Anaya
Enno Veerman
Ulla Mandel
Bruce Baum
Henrik Clausen
Inka Brockhausen
FONDECYT
Our Students
Paola Pérez
Eduardo Goicovich
Yoon-Jeoung Kwon
Patricia Ewert
Marianela Sánchez
María José Barrera
Rodrigo Pinto
Mónica Brito
Juan Cortés
Isabel Castro
Verónica Bahamondes
Hsiao Hsin Sung
Camilo Tapia
Denisse Sepúlveda
Hery Urra
José Velozo
Sebastián Indo
Katherine Bravo
Camilo Tapia
Sebastián Puga
Carolina Lagos`
Patricia Carvajal
Nicolás Albornoz
A heartfelt gratitude to our Patients
46. Model of the changes in assembly of type I-HD
in LSG from SS patients
González S et al ARTHRITIS & RHEUMATISM 63, 2011, 1106–1115