П. Сутерс "Проявления инсулинорезистентности и гликемический контроль в интен...rnw-aspen
Доклад с 15 Межрегиональной научно-практической конференции "Искусственное питание и инфузионная терапия больных в медицине критических состояний" 21-22 мая 2015 г
Alterations of Hepcidin and Interleukin in Diabeticsasclepiuspdfs
Aim: This study was done to determine the levels of hepcidin and interleukin-6 (IL-6) in diabetics. Materials and Methods: The study involved 100 diabetics and 100 apparently normal subjects of the same age group 40–70. The levels of hepcidin and IL-6 were measured with an enzyme-linked immunosorbent assay method. Results: The level of hepcidin and IL-6 significantly increased in the diabetics when compared to the controls (P < 0.05). Conclusion: This study probably indicated a significant increase in serum IL-6 and hepcidin levels in patients with diabetics.
Abo incompatible kidney transplantion - A single center experiencemionresearch
We report our experience in ABO incompatible transplantations done in 13 patients since 2009. The transplantations have been across different blood group combinations. The pre conditioning of the patient was done as per the Japanese protocol. Follow up period has been from 4 weeks to 28 months. 2 patients had immediate antibody mediated rejection with loss of graft. The rest 11 patients have normal graft function without any complications. Successful ABO incompatible transplantation is feasible in our country without endangering the life of recipient with reasonable cost control. Further studies are required to modify the protocol to prevent immediate antibody mediated rejections (ABMR).
П. Сутерс "Проявления инсулинорезистентности и гликемический контроль в интен...rnw-aspen
Доклад с 15 Межрегиональной научно-практической конференции "Искусственное питание и инфузионная терапия больных в медицине критических состояний" 21-22 мая 2015 г
Alterations of Hepcidin and Interleukin in Diabeticsasclepiuspdfs
Aim: This study was done to determine the levels of hepcidin and interleukin-6 (IL-6) in diabetics. Materials and Methods: The study involved 100 diabetics and 100 apparently normal subjects of the same age group 40–70. The levels of hepcidin and IL-6 were measured with an enzyme-linked immunosorbent assay method. Results: The level of hepcidin and IL-6 significantly increased in the diabetics when compared to the controls (P < 0.05). Conclusion: This study probably indicated a significant increase in serum IL-6 and hepcidin levels in patients with diabetics.
Abo incompatible kidney transplantion - A single center experiencemionresearch
We report our experience in ABO incompatible transplantations done in 13 patients since 2009. The transplantations have been across different blood group combinations. The pre conditioning of the patient was done as per the Japanese protocol. Follow up period has been from 4 weeks to 28 months. 2 patients had immediate antibody mediated rejection with loss of graft. The rest 11 patients have normal graft function without any complications. Successful ABO incompatible transplantation is feasible in our country without endangering the life of recipient with reasonable cost control. Further studies are required to modify the protocol to prevent immediate antibody mediated rejections (ABMR).
Stem Cell Research and Therapy - Cryoviva IndiaAnkita-rastogi
Stem cell therapy has recently been introduced to treat patients with type 2 diabetes mellitus (T2DM). However, no data are available on the efficacy and safety of allogeneic Whartons Jelly-derived mesenchymal stem cell (WJ-MSC) transplantation in patients with T2DM.
For more details visit at - http://www.cryoviva.in/banking-benefits/stem-cells-building-blocks-of-the-body/
Serum zinc, copper and iron in children with chronic liver diseasesNoha Lotfy Ibrahim
Trace elements play a major role as both oxidants and antioxidants, promoting and protecting from tissue damage. The liver plays a pivotal role in the metabolism of trace elements and consequently their bioavailability. Therefore, we aimed to measure serum levels of essential trace elements in children with chronic liver diseases (CLDs) and to study their correlation with liver function tests.
An update of this lecture is available at: https://www.slideshare.net/MohammedGawad/membranous-nephropathy-234601451
- Visit our website for more lectures: www.NephroTube.com
- Subscribe to our YouTube channel: www.youtube.com/NephroTube
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Increase your Understanding of the Pathogenesis of Gluten Spectrum DisordersCell Science Systems
Recently, researchers at Harvard University, Alessio Fasano et. al., and the National Institutes of Health (laboratories of immunology and cellular and molecular biology), reported real-time microscopic observations of gluten-induced neutrophil activation.
According to authors, " To what extent neutrophil function adds to, or protects against, gluten intolerance is currently under vigorous investigation."
This presentation will shed light on this question. It will also review the Fasano study and examine the role of neutrophil function in multiple disease conditions, as well as explore how neutrophil function may also play a dual role in protecting the body from the untoward effects of dietary and environmental agents.
Stem Cell Research and Therapy - Cryoviva IndiaAnkita-rastogi
Stem cell therapy has recently been introduced to treat patients with type 2 diabetes mellitus (T2DM). However, no data are available on the efficacy and safety of allogeneic Whartons Jelly-derived mesenchymal stem cell (WJ-MSC) transplantation in patients with T2DM.
For more details visit at - http://www.cryoviva.in/banking-benefits/stem-cells-building-blocks-of-the-body/
Serum zinc, copper and iron in children with chronic liver diseasesNoha Lotfy Ibrahim
Trace elements play a major role as both oxidants and antioxidants, promoting and protecting from tissue damage. The liver plays a pivotal role in the metabolism of trace elements and consequently their bioavailability. Therefore, we aimed to measure serum levels of essential trace elements in children with chronic liver diseases (CLDs) and to study their correlation with liver function tests.
An update of this lecture is available at: https://www.slideshare.net/MohammedGawad/membranous-nephropathy-234601451
- Visit our website for more lectures: www.NephroTube.com
- Subscribe to our YouTube channel: www.youtube.com/NephroTube
- Join our facebook group: www.facebook.com/groups/NephroTube
- Like our facebook page: www.facebook.com/NephroTube
- Follow us on twitter: www.twitter.com/NephroTube
Increase your Understanding of the Pathogenesis of Gluten Spectrum DisordersCell Science Systems
Recently, researchers at Harvard University, Alessio Fasano et. al., and the National Institutes of Health (laboratories of immunology and cellular and molecular biology), reported real-time microscopic observations of gluten-induced neutrophil activation.
According to authors, " To what extent neutrophil function adds to, or protects against, gluten intolerance is currently under vigorous investigation."
This presentation will shed light on this question. It will also review the Fasano study and examine the role of neutrophil function in multiple disease conditions, as well as explore how neutrophil function may also play a dual role in protecting the body from the untoward effects of dietary and environmental agents.
Diagnosis of Inflammatory bowel disease have challenges including differentiating from Irritable bowel disease using noninvasive biomarkers. Fecal calprotectin is a novel fecal marker which meets the diagnostic & monitoring requirements for IBD.
Just released to the public domain: Results from use of HepQuant technology in a study of Ledipasvir/Sofosbuvir HCV antivirals.. “Early Improvement in the HepQuant®(HQ)-SHUNT Function Test during Treatment with Ledipasvir/Sofosbuvir in Liver Transplant Recipients with Allograft Fibrosis or Cirrhosis and Patients with Decompensated Cirrhosis who have not undergone Transplantation. O’Leary JG, Burton JR, Helmke SM, Herman A, Cookson MW, Lauriski S, Trotter JF, Denning JM, Pang PS, McHutchison JG, Everson GT. Hepatology 2014;60:1134A.”
Similar to 23 jan final slides copy (dr pooja-vaio's conflicted copy 2013-12-14) (20)
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
MANAGEMENT OF ATRIOVENTRICULAR CONDUCTION BLOCK.pdfJim Jacob Roy
Cardiac conduction defects can occur due to various causes.
Atrioventricular conduction blocks ( AV blocks ) are classified into 3 types.
This document describes the acute management of AV block.
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
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
2. Background
Genetic • Autoimmune Disease
predisposition
(host factors)
Environmental
trigger
(exogenous
factors)
Immune
response
(host factors)
How this antigen enter?
Skin
Lung
Intestine
Eyes
Tissue damage
Amplified cycle of inflammation,
allow further leakage of foreign Ag
Th1
Th2
Foreign Ag entry
through barrier defect
APC
T cell
TNF-α
IFN-γ
3. Intestinal mucosal barrier system
• The extrinsic barrier consisting i.e. mucus, bicarbonate, hormones,
cytokines prostaglandins
• The intrinsic barrier is composed of the epithelial cells lining i.e.
Junctions and channels
• Paracellular pathway (junction)
• Transcellular pathway (channel)
Paracellular pathway regulated by 4 specific junction
• Tight Junction
• Adherens junction
• Desmosomes
• Gap junction
4. TJs regulate intestinal permeability
• Permeability is a process where molecules are allowed to
pass through the epithelial lining by non mediated diffusion
• Barrier properties of the intestinal epithelium are regulated by
TJs. It is generally believed that disease related, increase in
IP is caused by defects in TJ structure
No disease Disease
Normal TJ
regulate normal IP
Disrupted TJ
leads to increase IP
5. Why to study Tight Junction ?
PARACELLULAR
LLuummeenn
TRANSCELLULAR
TTiigghhtt
JJuunnccttiioonn
BBllooooddssttrreeaamm
Paracellular permeability between the epithelial
cells (gate function )
Maintenance of apical basolateral cell polarity
(fence function)
6. Tight Junction Proteins
Name of Proteins Known Partner
Claudin family ZO-1
Occludin ZO-1, ZO-2,Vap33, Actin
JAM family ZO-1, MUPP1
ZO family
ZO-1
ZO-2
ZO-3
Occludin, Claudin , ZO-2-3, Cingulin, Actin & ZONAB
ZO-1, Actin
ZO-1, Occludin
Ras, Zo-1
Tricellulin
AF-6
DLg (Drosophila)
Scribble(Drosophila)
Cingulin Occludin, ZO-1-2, myosin
Symplekin
ASIP/Par3 PKC-a
Rab3b
Rab13 d-PDE
Rab8 G/C Kinase, exocyst
Sec6, Sec8
8. Distribution of Tight Junctions
Villous
crypt
• At the tip of the villi the TJs are smaller in size but larger in number and
more accessible
• At the crypt the TJs are larger in size but less in number and less accessible
• Small molecules (mannitol , 6.7 Ao) = Villous
• Larger molecules (lactulose, 9.5 Ao) = Crypt
9. TJ proteins in celiac disease
↓ ZO-1 in celiac patients (n=10) which normalized 10 months
after treatment (Montalto 2002)
↓ ZO-1 in celiac patients (n=20) which reverse after
treatment along with increased IP (Pizzuti 2002)
↓ZO-1 and occludin , on exposure to gliadin in vivo and vitro,
zonulin signaling gets activated, leading to increased IP
(Drago 2006)
↓ZO-1 and occludin both in in vivo and vitro (Ciccocioppo
2006)
↑ of Claudin- 2 & claudin-3 in naïve celiac patients (n=33)
responsible for increased IP (Sapone 2011)
Alteration in TJ proteins leads to increased IP, which improves after
GFD( gluten free diet)
10. TJ proteins in Crohn’s disease
• ↓ of Occludin in colonic epithelial cells of active IBD( UC and
CD) play a role in enhanced paracellular permeability
(Kucharzik 2001)
• ↑ of Claudin- 2 and ↓ claudin 3 &4, which have different
effect of cytokines and leads to increased IP (Shyam Prasad
2005)
• ↑ of Claudin- 2 and ↓ claudin 5 &8 leads altered TJ structure
leads barrier dysfunction (Zeissig 2007)
• ↑ claudin-1 and claudin-2 expression may be involved at
early stages of transformation in IBD-associated
neoplasia( Weber 2008)
Alteration in TJ proteins along with increased IP
11. IP in celiac and Crohn’s disease
Increased IP has been implicated in the pathogenesis of Crohn’s
disease, celiac disease
Increased IP in celiac patients is 60-80%
Studies for increased IP in case of Crohn’s disease is 36-40%, while
our study is also reported as 36% abnormal IP in Crohn’s disease
patients
Abnormal IP in patients with celiac disease is because of increase
in mannitol, while high lactulose in those with Crohn’s disease
Vogelsang H, Oberhuber G, Wyatt J 1996, Benjamin J, Govind K Makharia 2008
12. Lacunae
Lack of literature on the differences in tight junction
proteins in patients with Crohn’s disease and celiac
disease
Lack of knowledge on whether there is a difference in
the expression of tight junction proteins in those with or
without increase in intestinal permeability
Lack of literature on the effect of treatment in the tight
junction proteins expression in Crohn’s disease and
celiac disease
13. Differential expression of
TJ proteins in CD and
celiac may be attributed to
TJs disintegrity
Leads to ↑ IP, implicated in
pathogenesis of CD and
celiac disease
Restoration of TJP & IP,
may improve course of
CD and Celiac disease
Hypothesis
14. Objectives
Primary objective
To study the expression of tight junction proteins (Zo-1, Claudin 2, 3
& 4, Occludin and JAM) using Immunohistochemistry in patients
with Crohn's disease, celiac disease and controls
To study the ultrastructural changes of tight junction and to note the
changes of other junctions of a paracellular pathway by electron
microscopy
Secondary objective
To study the effect of treatment on the expression pattern of tight
junction proteins examined, as well as to see if any ultrastructural
improvement has taken place post-treatment, in those with celiac
disease and Crohn's disease
16. Patients
Crohn’s disease
• Diagnosis
ECCO guidelines
• Activity
CDAI score
• Location & Behavior
Montreal classification
Celiac disease
• Diagnosis
ESPGHAN criteria
• Pathologic changes (Activity)
Modified Marsh classification
ECCO, Gut, 2006
Best et al 1976
Silverberg et al 2005
ESPGHAN, Arch Dis Child 1990
Oberhuber 1999
17. Study criteria
Disease Inclusion criteria Exclusion criteria
Crohn’s disease 1. Age: 12 to 65 years
2. Both genders
3. Active disease
(Crohn’s disease
activity index >150-450)
1. Severely sick patients
2. Severe active disease (CDAI>450)
3. Massive bleeding
4. Those on active NSAID use
5. Patients with small intestinal resection
6. Ulcerative colitis
7. Indeterminate colitis
8. Intestinal tuberculosis
9. Patients with chronic or acute renal failure
10. Patients with intestinal perforation
11. Subject with other concomitants diseases
12. Pregnant and lactating mothers
13. Unwilling patients
Celiac disease 1. Age: 12 to 65 years
2. Both genders
3. Treatment Naïve
1. Patients who have received gluten free diet
earlier
2. Those on active NSAID use
3. Subject with other concomitants diseases
4. Pregnant and lactating mothers
5. Unwilling patients
6. Patients with tropical sprue
18. Selection of controls
Patients with functional dyspepsia, age (12-65)
of both gender served as controls, with no
alarming signal
Diarrhea
Anemia
Normal liver function test
Use of NSAID 15 days prior to study
Negative for celiac serology
Normal UGI endoscopy and normal histology
19. Work Algorithm
Patient with Crohn’s disease, celiac disease and controls recruited from the
Gastroenterology OPD of AIIMS
Inclusion and exclusion criteria
Informed consent taken
Diagnosis
•ECCO guideline for Crohn’s disease
•ESPGHAN criteria for celiac disease
Biochemical test, IP test, UGI endoscopy followed by Bx in CD, celiac and Controls
Duodenal Biopsy
Immunohistochemistry
•ZO-1
•Cld 2,cld-3 & cld-4
•Occludin,
• JAM
Histopathology Electron Microscopy
After 6 month follow up by clinician, we repeated all the
test in patients with celiac and Crohn’s disease
TJ protein
expression
Ultrastructural
changes
Effect of treatment
Objectives
20. Patients Enrollment
Crohn’s disease
Screened (n=50)
Excluded (n= 22 )
•Over age (n=1)
•Active bleeding (n=11)
•Co morbidity (n=5)
•Refused to participate (n=5)
Included (n=28)
Active Crohn’s disease
Follow-Up
Lost to follow up=8
Post treatment (n=20)
Celiac disease
Screened (n=80)
Excluded (n= 56 )
• Already on GFD (n=20)
• Co morbidity (n=31)
• Refused to participate (n=5)
Included (n=24)
Treatment naïve celiac
Follow-Up
Lost to follow up=3
Post treatment (n=21)
21. Analysis TJ structure and TJ protein
• TJ protein expression
– Immunohistochemistry, RT PCR, Western blotting
• Functional analysis of TJ
– IP by High performance liquid chromatography (HPLC), Ussing
chamber
• Ultrastructural of TJ
– Transmission electron microscopy (TEM), freeze fracture
Celiac, Crohn’s disease and
controls
TJ protein expression
Immunohistochemistry
(IHC)
Functional analysis of TJ
High performance liquid
chromatography(HPLC)
Ultrastructure of TJ by
Transmission electron
microscopy (TEM)
After 6 month test only in Celiac, Crohn’s disease
22. Statistical test
STATA 11.0 (College Station Texas USA) software used
Qualitative expression of TJ protein done by Fisher Exacts Test
Quantitative parameters of TEM and HPLC, by one way ANOVA
followed by Boneforroni for multiple regression
The effect of treatment assessed by using Mc Nemar’s test
Pairwise correlation coefficient for correlation
p value of <0.05 was considered statistically significant
23. Primary objective
To study the expression of tight junction proteins
(Zo-1, Claudin 2, 3 & 4, Occludin and JAM)
using Immunohistochemistry in patients with
Crohn's disease, celiac disease and controls
24. Immunohistochemistry Methodology
ZO-1 1:20
Claudin-2 1:50
Claudin-3 1:40
Claudin-4 1:400
JAM-A 1:40
occludin 1:20
Villous
Crypt
MS
ICJ
Cytoplasm
Nucleus
Distribution
Intensity
Negative 0
V. Focal <20% surface area
Focal >21%-50% surface area
Diffuse >50% surface area
Negative- 0
Faint 1
Moderate- strong 2
Strong 3
Villous
Crypt
Mucosal Surface (MS)
Intracellular junction
(ICJ)
Cytoplasm
Nucleus
25. Immunohistochemistry two day protocol
Steps Methods
Tissue Paraffinized section of duodenal biopsy
Deparaffinization Deparaffinized the slide in xylene and followed by acetone
& ethanol washing with running tap water
Blocking Blocking in 0.3% H2O2 for 30 minute washing in x PBS 3
times
Retrieval Prewarm then retrieval treatment
Primary Ab( Zymed, San
Francisco, CA) binding
Primary antibody binding for overnight incubation at 40 C
Secondary Ab (DAKO, Envision
real system) binding
After 3 washing x PBS incubate with secondary for 1
hour.
Colour development Di- aminobenizidine tetra hydrochloride (DAB)
Washing To stop the reaction, we washed the slides in water
Counterstaining By using hematoxylin stain
Mounting DPX Mount
Detection Examined using light microscope
26. TJ protein expression in normal control biopsy
villous vs. crypt
Villi Crypt
Claudin-2
Villi Crypt
Claudin-3
Villi Crypt
ZO-1
Villi Crypt
JAM
Villi Crypt Villi Crypt
Occludin Claudin-4
27. ZO-1 expression in celiac and Crohn’s disease
control
Treatment
naïve celiac
Active Crohn’s
disease
ZO-1 expression in villous and
crypt
Control vs
treatment naïve
celiac (P)
Control vs. Active
Crohn’s disease
(P)
Celiac vs. Crohn’s
(P)
Distribution in villous mucosal
surface
0.02 0.03 0.29
Intensity in villous mucosal surface 0.003 0.03 0.13
Distribution in crypt mucosal surface 0.05 0.008 0.84
Intensity in crypt mucosal surface 0.02 0.02 1.0
28. Cld-2 expression in Celiac and Crohn’s disease
Control
Treatment
naïve celiac
Active Crohn’s
disease
Cld-2 expression in villous and crypt Control vs
treatment naïve
celiac (P)
Control vs. Active
Crohn’s disease
(P)
Celiac vs. Crohn’s
(P)
Distribution in villous ICJ 0.002 0.001 0.27
Intensity in villous ICJ 0.02 0.03 0.03↑ in CD
Distribution in crypt ICJ 0.001 0.007 0.70
Intensity in crypt ICJ 0.01 0.001 0.73
29. Claudin-3 expression in Celiac and Crohn’s disease
Treatment
naïve celiac
Active Crohn’s
disease
Control
Cld-3expression in villous and crypt Control vs
treatment naïve
celiac (P)
Control vs. Active
Crohn’s disease
(P)
Celiac vs. Crohn’s
(P)
Distribution in villous ICJ 0.003 0.78 0.008 in CD
Intensity in villous ICJ 0.17 0.64 0.03 in CD
Distribution in crypt ICJ 0.01 0.43 0.009 in CD
Intensity in crypt ICJ 0.26 0.33 0.17
30. Claudin-4 expression in Celiac and Crohn’s disease
Control
Cld-4expression in villous and crypt Control vs
treatment naïve
celiac (P)
Control vs. Active
Crohn’s disease
(P)
Celiac vs. Crohn’s
(P)
Distribution in villous ICJ 0.08 0.29 0.04↑ in CD
Intensity in villous ICJ 0.18 0.44 0.32
Distribution in crypt ICJ 0.10 0.18 0.07
Intensity in crypt ICJ 0.24 0.20 0.28
Treatment
naïve celiac
Active Crohn’s
disease
31. Occludin expression in Celiac and Crohn’s disease
Control
Treatment
naïve celiac
Active Crohn’s
disease
Occ expression in villous and
crypt
Control vs treatment
naïve celiac (P)
Control vs. Active
Crohn’s disease
(P)
Celiac vs. Crohn’s
(P)
Distribution in villous ICJ <0.01 <0.01 0.66
Intensity in villous ICJ <0.01 <0.01 0.06
Distribution in crypt ICJ <0.01 <0.01 0.15
Intensity in crypt ICJ <0.01 <0.01 0.74
32. JAM expression in Celiac and Crohn’s disease
Control
JAM expression in villous and crypt Control vs
treatment naïve
celiac (P)
Control vs. Active
Crohn’s disease
(P)
Celiac vs. Crohn’s
(P)
Distribution in villous ICJ 0.21 1.0 0.05
Intensity in villous ICJ 0.39 0.07 0.30
Distribution in crypt ICJ 0.76 0.58 0.53
Intensity in crypt ICJ 0.15 1.0 1.0
Treatment
naïve celiac
Active Crohn’s
disease
33. Functional analysis of TJ
Assessment of IP by dual sugar
permeation test(LM ratio)
Lactulose mannitol ratio estimated
in urine sample by HPLC
Pumping unit
Sample-injection unit(injector)
Separation unit ( Column)
Detection unit (ELSD)
Data-processing unit
35. ELSD (Evaporating light scattering detector)
• Nebulisation of the effluent: Transformation of solvent from the
HPLC column into fine droplets
• Evaporation of the effluent: Droplets are carried by the gas flow
into the heated area
• Detection: The sample particles pass through a flow cell where
they get detect
36. Protocol for HPLC
Standard Urine sample
Serial dilution of standard i.e. lactulose,
Mannitol & cellobiose (75μg-5μg)
Conditioning by Maxi-Clean C18 600-mg
cartridges with 5 ml of methanol followed
by 5 ml of water
Passed the sugars in conditioned C-18
cartridge, residual volume was collected
and diluted 1:1 with water (cellobiose and
amberlite IRA-400) vortex for 10 s and
centrifuged for 2 min at 3000 rpm
Filter the supernatant in syringe filters
(Nylon 66, 0.2 mm) & centrifuge for 5 min
at 3000 rpm
Analyzed the standard by ELSD-HPLC
Centrifuge the urine (30%TCA) sample at
3000 rpm for 10 min
Conditioning by Maxi-Clean C18 600-mg
cartridges with 5 ml of methanol followed by
5 ml of water
Passed the urine from C-18 cartridge,
residual volume was collected and diluted
1:1 with water (cellobiose and amberlite
IRA-400) vortex for 10 s and centrifuged for
2 min at 3000 rpm
Filter the supernatant in syringe filters
(Nylon 66, 0.2 mm) ¢rifuge for 5 min at
3000 rpm
Analyzed the sample by ELSD-HPLC
37. Standard chromatogram
HPLC and ELSD components Standardized condition
Mannitol
Column 250 ´ 4.6 mm column
Mobile phase Water & Acetonitrile (ACN)
Mobile phase flow rate 1.0 ml/min
ELSD detector temperature 650C
ELSD flow rate 2 litre/min
Column temperature 400C
Lactulose
Cellobiose
Mannitol Retention time 7.3 minute
Lactulose Retention time 8.3 minute
Cellobiose Retention time 9.6 minutes
Total Retention time 12 minutes
Injector volume 10μl with needle wash
38. Calibration graph of urine and water
Urine Water
Concentration Concentration
Recovery = Concentration of (L or M) in Urine ×100
Concentration of (L or M) in water
Recovery was (97%-100%)
Lower detection limit for lactulose is 6.25mg/L and 3.25mg/L for mannitol
Area
Area
39. Intestinal permeability in celiac and
Crohn’s disease
Control Treatment naïve
celiac
P
Lactulose % 0.002±.001 0.004±.003 0.013↑
Mannitol % 0.04±.02 0.024±.013 0.037↓
LMR 0.07±.04 0.19±0.19 0.006↑
Control Active Crohn’s
disease
P
Data in mean ±SD
Lactulose % 0.002±.001 0.005±.002 0.0005↑
Mannitol % 0.040±.02 0.024±.019 0.038↓
LMR 0.07±.04 0.22±.16 <.01↑
Data in mean ±SD
40. Primary Objective
To study the ultrastructural changes of tight
junction and to note the changes of other
junctions of a paracellular pathway by electron
microscopy
41. Transmission Electron Microscope
To increase resolution of the
object many-folds over
conventional light microscope
Image analyses were performed
to measure
TJ diameter,
Microvilli (MV) length
Inter-MV width
Mitochondrial diameters (MD)
42. Sample preparation for TEM
Steps Description
Primary fixation Very small 1-2mm2 thick duodenal fragments were directly fixed in 2.5%
gluteraldehyde, made in 0.1M sodium phosphate buffer (pH=7.4).
Washing After fixation, repeated washing was made in 0.1M sodium phosphate
buffer (pH=7.4).
Post fixation OsO4(1% solution) was used as the secondary fixative, and it preserves
the lipid and again washed in 0.1M sodium phosphate buffer (pH=7.4).
Dehydration As the tissue dehydrated by passing through a series of ascending
concentration of ethanol
Clearing Epoxy propane was used for removing the traces of water in case of
incomplete dehydration.
Embedding Embedding was done in embedding capsule using gelatine or beam
capsule.
Polymerization To polymerize tissue, kept at 400 c - 500 c for overnight. After
polymerization capsules were washed in water at 700 c.
43. Continued
Steps Description
Staining for LM The semithin sections (thick 0.5 -2μm) was then stained with toludine
blue. After cleaning the stained section they were observed under light
microscope to mark the area of interest and went for ultra thin section
cutting
Staining for EM with
Uranyl acetate
Staining of ultrathin section ( thick 70-80 nm) was done with 50μl of
uranyl acetate
Staining with
Lead acetate
After washing of grid in double distilled water and dried carefully on a
filter paper. And stain with Lead acetate and lead citrate inside a
petridish.
Washing The grids were then washed in 0.02 M NaOH, followed by twice in
double distilled water. The grids were then dried carefully.
Acquiring of images The stained sections were then observed under a Morgagni 268D
TEM at an operating voltage 80 kV. Images were digitally acquired by
using a CCD camera (Megan view III, Fei Company) attached to the
microscope
45. Pentalaminar structure of Tight Junction
TJ
AJ
Desmosomes
Gap junction
Pentalaminar structure
X 120000
46. Morphometric measurement of TJ and other
ultrastructures
25000 x
Tight junction Mitochondria
10000 x
5000 x 10000 x
Microvillous length Intervillous width
47. Ultrastructural changes in celiac and Crohn’s
disease
Characteristics in Ultra
structure
Control
n=5(%)
Treatment naïve
Celiac n=12(%)
P
Alternation of TJ 0 5(41.6) 0.6
Pentalaminar structure 0 3(25) 0.3
ICJ 0 3(25) 0.3
Desmosomes 0 4(33.3) 0.2
Gap junction 0 0(0) 1.0
Mitochondria dilation 0 6(50) 0.2
Microvilli disarray 0 4(33.3) 0.2
Characteristics in Ultra
Control
Active Crohn’s
structure
n=5(%)
disease n=10(%)
P
Alternation of TJ 0 6(60) 0.1
Pentalaminar structure 0 6(60) 0.1
ICJ 0 2(20) 0.3
Desmosomes 0 5(50) 0.0
Gap junction 0 0(0) 1.0
Mitochondria dilation 0 2(20) 0.4
Microvilli disarray 0 3(30) 0.2
48. Ultrastructural quantitative changes of celiac
disease compare to control
TJ morphology Control
(n=5)
Treatment naïve
celiac(n=12)
P
Control
Diameter of TJ(nm) 21.67±20.6 31.14±21.6 0.5
Inter villous width(nm) 94.47±27.0 156.77±95.2 0.4
Length of microvilli(nm) 1250.31±343.9 1075±321.5 0.9
Diameter of Mitochondria(nm) 741.60±238.7 784.86±200.3 0.7
Treatment naïve
celiac
Tight junction Mitochondria Microvillous length Desmosome
49. Ultrastructural quantitative changes of Crohn’s
disease compare to controls
RESULTS
TJ morphology Control
(n=5)
Active CD
(n=10)
P
Control
Diameter of TJ(nm) 21.67±20.6 45.10±32.1 0.8
Inter villous width(nm) 94.47±27.0 140.35±77.5 0.7
Length of microvilli(nm) 1250.31±343.9 1110±436.2 0.7
Diameter of Mitochondria (nm) 741.60±238.7 826.77±236.4 0.3
Active Crohn’s disease
Tight junction Mitochondria Microvillous length Desmosome
50. Secondary objective
To study the effect of treatment on the
expression pattern of tight junction proteins
examined, as well as to see if any ultrastructural
improvement has taken place post-treatment, in
those with celiac disease and Crohn's disease.
54. Histological changes in celiac disease 6 month
after treatment
Post treatment
Pre Rx 0(N) 1(N) 2(3a) 3(3b) 4(3c) Total P
0 0 0 0 0 0 0
1 0 0 0 0 0 0
2(3a) 1 0 0 0 0 2 0.009
3(3b) 0 0 2 4 0 6
4(3c) 0 3 2 3 6 14
Total 1 4 4 7 6 22
55. Disease activity and Histological change in Crohn’s
disease after treatment
Variable Pre treatment
(n=28)
Post-treatment
(n=20)
P
CDAI Score 250.95±65.07 140.24±81.05
Remission (£150) 0(0%) 11(56.54%) 0.0005
Mild (151-220) 13(43.33%) 6(21.73%)
Moderate (221-400) 17(56.67%) 3(21.73%)
Severe (>400) 0(0%) 0(0%)
Post treatment
Pre Treatment 0(N) 2(3a) 3(3b) 4(3c) Total P
0 (N) 9 0 1 0 10
0.31
2(3a) 2 1 0 2 5
3(3b) 2 0 1 0 3
4(3c) 1 0 1 0 2
Total 14 1 3 2 20
56. Celiac pre vs post
(P)
ZO-1 in celiac pretreatment
Distribution in villous mucosal surface 0.02 0.02
Intensity in villous mucosal surface 0.003 0.03
Distribution in crypt mucosal surface 0.05 0.34
Intensity in crypt mucosal surface 0.02 0.75
ZO-1 in Celiac post-GFD
ZO-1 in CD pre-treatment
ZO-1 in CD post-treatment
ZO-1 expression in villous and crypt Control vs Celiac
(P)
ZO-1 expression in villous and crypt Control vs. Crohn’s
(P)
Crohn’s disease pre vs post
(P)
Distribution in villous mucosal surface 0.3 0.04
Intensity in villous mucosal surface 0.03 0.13
Distribution in crypt mucosal surface 0.008 0.14
Intensity in crypt mucosal surface 0.02 0.12
57. Cld -2 Celiac disease pre-treatment
Cld-2 expression in villous and crypt Control vs Celiac
Celiac pre vs post
(P)
Distribution in villous ICJ 0.002 0.12
Intensity in villous ICJ 0.02 0.14
Distribution in crypt ICJ 0.001 0.71
Intensity in crypt ICJ 0.01 0.55
Cld-2 Celiac disease post treatment
Cld -2 Crohn’s disease pre-treatment
Cld-2 Crohn’s disease post treatment
a b c
(P)
Cld-2 expression in villous and crypt Control vs. CD
(P)
CD pre vs post
(P)
Distribution in villous ICJ 0.001 1.0
Intensity in villous ICJ 0.03 0.25
Distribution in crypt ICJ 0.007 0.25
Intensity in crypt ICJ 0.001 0.37
58. Cld -3 Celiac disease pre-treatment
Cld-3 Celiac disease post treatment
Cld -3 Crohn’s disease pre-treatment
Cld-3 Crohn’s disease post treatment
Cld-3expression in villous and crypt Control vs Celiac
(P)
Celiac pre vs post
(P)
Distribution in villous ICJ 0.003 0.59
Intensity in villous ICJ 0.17 0.19
Distribution in crypt ICJ 0.01 0.45
Intensity in crypt ICJ 0.26 1.0
Cld-3expression in villous and crypt Control vs CD
(P)
CD pre vs post
(P)
Distribution in villous ICJ 0.78 1.0
Intensity in villous ICJ 0.64 1.0
Distribution in crypt ICJ 0.43 1.0
Intensity in crypt ICJ 0.33 1.0
59. Cld-4expression in villous and crypt Control vs Celiac
Cld -4 Celiac disease pre-treatment
Cld-4 Celiac disease post treatment
Cld -4 Crohn’s disease pre-treatment
Cld-4 Crohn’s disease post treatment
(P)
Celiac pre vs post
(P)
Distribution in villous ICJ 0.08 0.57
Intensity in villous ICJ 0.18 0.3
Distribution in crypt ICJ 0.10 0.7
Intensity in crypt ICJ 0.24 0.23
Cld-4expression in villous and crypt Control vs. CD
(P)
CD pre vs post
(P)
Distribution in villous ICJ 0.29 0.88
Intensity in villous ICJ 0.44 0.57
Distribution in crypt ICJ 0.18 0.59
Intensity in crypt ICJ 0.20 0.43
60. Occludin Celiac disease pre-treatment
Occ expression in villous and crypt Control vs Celiac
Occludin Celiac disease post treatment
Occludin Crohn’s disease pre-treatment
Occludin Crohn’s disease post treatment
(P)
Celiac pre vs post
(P)
Distribution in villous ICJ <0.01 1.0
Intensity in villous ICJ <0.01 1.0
Distribution in crypt ICJ <0.01 0.5
Intensity in crypt ICJ <0.01 1.0
Occ expression in villous and crypt Control vs. CD
(P)
CD pre vs post
(P)
Distribution in villous ICJ <0.01 0.6
Intensity in villous ICJ <0.01 0.3
Distribution in crypt ICJ <0.01 0.25
Intensity in crypt ICJ <0.01 0.05
61. JAM Celiac disease pre-treatment
JAM expression in villous and crypt Control vs Celiac
JAM Celiac disease post treatment
JAM Crohn’s disease pre-treatment
JAM Crohn’s disease post treatment
(P)
Celiac pre vs post
(P)
Distribution in villous ICJ 0.2 1.0
Intensity in villous ICJ 0.39 1.0
Distribution in crypt ICJ 0.76 1.0
Intensity in crypt ICJ 0.15 1.0
JAM expression in villous and crypt Control vs. CD
(P)
CD pre vs post
(P)
Distribution in villous ICJ 1.0 1.0
Intensity in villous ICJ 0.07 0.21
Distribution in crypt ICJ 0.58 0.5
Intensity in crypt ICJ 1.0 0.25
62. Effect of treatment on IP in celiac and Crohn’s
disease
Treatment naïve celiac
(n=24)
Celiac after GFD
(n=21)
P
Lactulose % 0.004±.003 0.002±0.001 0.01↓
Mannitol % 0.024±.013 0.020±0.007 0.18
LMR 0.19±0.19 0.12±0.06 0.04↓
Active CD
(n=28)
CD Post-treatment
(n=20)
P
Data expressed in mean± SD
Lactulose % 0.005±.002 0.002±0.001 0.002↓
Mannitol % 0.024±.019 0.020±0.019 0.48
LMR 0.22±.16 0.14±0.08 0.01↓
Data expressed in mean± SD
63. Ultrastructural changes in celiac disease 6
month after GFD
TJ morphology Treatment
Tight junction Desmosome Mitochondria Intervillous width
naïve celiac
(n=12)
Celiac after
GFD
(n=12)
P
Diameter of TJ(nm) 31.14±21.6 23.96±8.4 0.14
Pre-treatment
10000x
Pre-treatment
8000x
Pre-treatment
16000x
Pre-treatment
4000x
There was no correlation of the ultrastructural changes
Treatment naïve celiac
Inter villous width(nm) 156.77±95.2 164.28±124.2 0.87
Post-treatment
10000x
with changes in IP (LMR)
Post-treatment
8000x
Post-treatment
16000x
Post-treatment
4000x
Length of microvilli(nm) 1075±321.5 1110.89 ± 337.6
0.97
Diameter of Mitochondria (nm) 784.86±200.3 892.11± 173.3 0.85
Celiac disease post GFD
64. Effect of treatment on ultrastructural changes in
Crohn’s disease
TJ morphology Active CD
Tight junction Desmosome Mitochondria Intervillous width
(n=10)
CD Post-treatment(
n=10)
P
Diameter of TJ(nm) 45.10±32.1 20.37±7.2 0.04
Pre-treatment
10000x
Pre-treatment
3200x
Except from the inter-villous width, Pre-with treatment
lactulose excretion Pre-treatment
(p-0.01
& Inter r-0.72) villous ,there width(nm) was no correlation 140.35±77.5 8000x
of the ultrastructural 142.97±73.4 4000x
changes
0.94
Active Crohn’s disease
Post treatment
10000x
with IP changes( LMR)
Post treatment
3200x
Post treatment
8000x
Post treatment
4000x
Length of microvilli(nm) 1110±436.2 1506.84±1439
0.46
Diameter of Mitochondria(nm) 826.77±236.4 715.45±210.1 0.04
Crohn’s disease post-treatment
65. Summary: Celiac disease
Celiac disease vs. controls Celiac disease pre vs. post
Claudin-2
Claudin-3
Claudin-4
Occludin
ZO-1
ZO-1
JAM
IP Increased
Dilated TJ
Claudin-2
Claudin-3
Claudin-4
Occludin
ZO-1
ZO-1
JAM
IP improved
Dilated TJ
66. Summary: Crohn’s disease
Crohn’s disease vs. controls Crohn’s disease pre vs. post
Claudin-2
Claudin-3
Claudin-4
Occludin
ZO-1
ZO-1
JAM
IP Increased
Dilated TJ
Claudin-2
Claudin-3
Claudin-4
Occludin
ZO-1
ZO-1
JAM
IP improved
TJ improved
67. Summary: Crohn’s disease & celiac disease
Celiac vs. Crohn’s disease pre Rx Celiac vs. Crohn’s disease post Rx
Claudin-2
Claudin-3
Claudin-4
Occludin
ZO-1
ZO-1
JAM
IP , No change
Dilated TJ,
No change
Claudin-2
Claudin-3
Claudin-4
Occludin
ZO-1
ZO-1
JAM
IP, No change
Dilated TJ,
No change
68. Conclusion
Pretreatment
TJ proteins
Celiac disease
pore forming protein: ↑cld-2
Pore sealing protein : ↓ cld-3
Scaffold protein : ↓ ZO-1
Ultrastructure TJ dilated
IP Increase
Post-treatment
TJ proteins
pore forming protein : ↓ cld-2
Scaffold protein : ↑ ZO-1
Ultrastructure TJ dilated
IP Reduced
Crohn’s disease
Pretreatment:
TJ proteins
pore forming protein : ↑cld-2
Scaffold protein : ↓ ZO-1
Ultrastructure TJ dilated
IP Increase
Post-treatment:
TJ proteins
pore forming protein : ↓ cld-2
Scaffold protein : ↑ ZO-1
Ultrastructure partially resolved
IP Reduced
There was no difference in TJ protein expression, IP and TJ dilation in celiac
and Crohn’s disease at baseline as well as 6 month after treatment
69. Conclusion
Possibly the expression patterns of TJ proteins
- ZO-1
- pore forming protein claudin-2
- and pore sealing protein claudin-3
regulate the tight junction structure and intestinal
permeability
70. CHIEF GUIDE
Dr Govind K. Makharia
Additional Professor
Dept. of Gastroenterology & HNU
DC member CO-Guide
AIIMS, New Delhi
Dr. Vineet Ahuja
Additional Professor
Dept. of Gastroenterology & HNU
AIIMS, New Delhi
Dr S.K. Acharya
Professor & Head
Dept. of Gastroenterology & HNU
AIIMS, New Delhi
Dr Subroto Sinha
Professor & Head
Department of Biochemistry
AIIMS, New Delhi
Dr. S. k. Panda
Professor & Head
Dept. of Pathology
AIIMS, New Delhi
Dr. H.K. Prasad
Professor
Dept. of Biotechnology
AIIMS, New Delhi
Dr Y.K. Joshi
Professor
Dept. of Gastroenterology & HNU
AIIMS, New Delhi
Shyam Prakash
Scientist
Dept. of Gastroenterology & HNU
AIIMS, New Delhi
Dr. V. Sreenivas
Associate Professor
Dept. of Biostatistics
AIIMS, New Delhi
Dr. Siddhartha Datta Gupta
Professor
Dept. of Pathology
AIIMS, New Delhi
71. Awards
ISG WIN Medicare Award for Best oral Paper presentation :
P. Goswami*1, P. Das2, S. P. Khanel3, V. Sreenivas3, S. Datta
Gupta2, G. K. Makharia. “Expression of tight junction protein (ZO-1,
Claudin-2, 4 and Occludin) in patients with celiac disease and active
Crohn’s disease and change in their expression 6 months after
treatment” in ISG 2009 held at Kolkata
Prof K.C. Basumallick Award for best research paper : P Das,
Pooja Goswami, Siddharth Datta Gupta, Subrat K Panda & Govind
K Makharia, Tight Junction proteins (Zo-1, Claudin 2, 3 & 4,
Occludin and JAM) expression and Intestinal permeability in
patients with Celiac and Crohn's disease, and their association with
light microscopic and ultrastructural findings in APCON 2011 held at
Patiala
72. Publications Journal publication
Prasenjit Das, Pooja Goswami, Tapash K Das, Tapas Nag, Vishnubhatla Sreenivas, Vineet
Ahuja,Subrat K Panda, Siddhartha Datta Gupta, Govind K Makharia.. Comparative Tight junction
protein expressions in colonic Crohn’s disease, ulcerative colitis and tuberculosis: a new
perspective: Virchows Archiv 2012
Sood A, Midha V, Makharia GK, Ahuja V, Singal D, Goswami P, Tandon RK. . Probiotics for
ulcerative colitis ... Are the good bugs back? Gastroenterology, 2010 Sep; 139(3):1054-6. [Epub
ahead of print] PubMed PMID: 20667486.
Makharia GK, Srivastava S, Das P, Goswami P, Singh U, Tripathi M, Deo V, Aggrawal A, Tiwari
RP, Sreenivas V, Gupta SD. Clinical, endoscopic, and histological differentiations between
Crohn's disease and intestinal tuberculosis. Am J Gastroenterology, 2010 Mar; 105(3):642-51.
Epub 2010
G. K Makharia, A. Seth, S. K. Sharma, A. Sinha, P. Goswami, A. Aggrawal, K. Puri & V.
Sreenivas. Structural and functional abnormalities in lungs in patients with achalasia.
Neurogastroenterol Motility .2009 1365-2982.
Sood A, Midha V, Makharia GK, Ahuja V, Singal D, Goswami P, Tandon RK. The Probiotic
Preparation, VSL#3, Induces Remission in Patients With Mild-to-Moderately Active Ulcerative
Colitis. Clin Gastroenterol Hepatol. 2009;(11):1202-9
Das CJ, Makharia G, Kumar R, Chawla M, Goswami P, Sharma R, Malhotra A. PET-CT
enteroclysis: a new technique for evaluation of inflammatory diseases of the intestine. Eur J Nucl
Med Mol Imaging. 2007 ;(12):2106-14.
Gupta AK, Chauhan DS, Srivastava K, Das R, Batra S, Mittal M, Goswami P, Singhal N, Sharma
VD, Venkatesan K, Hasnain SE, Katoch VM. Estimation of efflux mediated multi-drug resistance
and its correlation with expression levels of two major efflux pumps in mycobacteria. J Commun
Dis. 2006 Mar; 38(3):246-54.
Book-Chapter:
Pooja Goswami, Chandan Jyoti, Govind K Makharia .Inflammatory bowel disease ; Update
2008.Non-invasive Diagnostic Tools for Inflammatory Bowel Disease: 32-35..Kolkata
Gastroenterology society on eve of Enterocon 2008.
73. Abstracts
P. Goswami*1, P. Das2, TC Nag, TK Das V. Sreenivas3, S. Datta Gupta2, G. K. Makharia. Tight junction alteration in
Celiac disease and Crohn’s disease and effect of treatment on them. J. of Gastroenterology and Hepatology 2010.
(584small intestine)
P. Goswami*1, Meenakshi Sharma, P. Das2, S. Datta Gupta2, G. K. Makharia. Difference in the features of celiac disease
in adolescents and adults. J. of Gastroenterology and Hepatology 2010.(582 small intestine)
P. Das2, G. K. Makharia, P. Goswami*1, , TC Nag, TK Das V. Sreenivas3, S. Datta Gupta2. Expression of tight junction
proteins in Crohn’s disease , ulcerative colitis and intestinal tuberculosis. – 2009 (585 large intestine)
P. Goswami*1, P. Das2, S. P. Khanel3, V. Sreenivas3, S. Datta Gupta2, G. K. Makharia. “Changes in the expression of
ZO-1, claudin-2, claudin-4 and Occludin in the duodenal mucosa of patients with celiac disease before and after treatment
with Gluten free diet”. J. of Gastroenterology and Hepatology 2009, P0079
S. Srivastava*1, G. K. Makharia1, P. Das2, P. Goswami1, S. Datta Gupta2 J.Development of a therapeutic algorithm for
management of intestinal tuberculosis and Crohn’s disease in tuberculosis endemic countries. of Gastroenterology and
Hepatology 2009,OP412
, U. Sharma1, R. R. Singh1, P. Goswami*2, V. Ahuja2, G. K. Makharia2, N. R. Jagannathan. Similarity in the metabolic
profile in macroscopically involved and uninvolved colonic mucosa in patients with inflammatory bowel disease: An in-vitro
proton MR. Spectroscopy study J. of Gastroenterology and Hepatology 2009, P1744
P. Goswami*2, Das C.J.,V. Ahuja2, Rakesh Kumar G. K. Makharia2 PET-CT Colonography: A novel non-invasive
technique for assessment of extent and activity of the disease in patients with ulcerative colitis. Gastroenterology and
Hepatology 2009, P1059
P. Goswami*1, P. Das2, S. P. Khanel3, V. Sreenivas3, S. Datta Gupta2, G. K. Makharia Expression of tight junction protein
(ZO-1, Claudin-2, 4 and Occludin) in patients with celiac disease and active Crohn’s disease and change in their expression
6 months after treatment Indian J.Gastroenterol.2009.
Pooja Goswami, Namrata Singh, Rajesh Khadgawat,* Siddhartha Datta Gupta,** Govind K Makharia. Response to
treatment in adult patients with celiac disease. J. of Gastroenterology and Hepatology 2008:23 Suppl.5):A-93
S. Srivastava, G.K. Makharia, U Singh, P Goswami, M. Tripathi, SD Gupta, YK Joshi. Development of therapeutic algorithm
of intestinal tuberculosis endemic area. J. of Gastroenterology and Hepatology 2008:23 Suppl.5): A-95.
Chandan J Das, Govind Makharia*, Raju Sharma, Pooja Goswami*, Rakesh Kumar, AK Malhotra. PET-CT Colonography:
A Novel Noninvasive Technique for Assessment of disease extent in Ulcerative Colitis. J. of Gastroenterology and
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