This document discusses diagnostic procedures for schistosomiasis in both high and low endemic areas. It describes several diagnostic methods including microscopic examination of stool, urine, or biopsy samples to detect parasite eggs. Serological tests to detect antibodies or antigens are also used when egg detection is difficult. These include ELISA, western blot, and dipstick tests to detect circulating cathodic antigen. Molecular tests can also detect parasite DNA from different life cycle stages. Direct microscopic examination remains the gold standard, but serology and molecular tests provide increased sensitivity for low-level infections.
LUMEN DWELLING FLAGELLATES - GIARDIA
REFS:
INTERNATIONALLY ACCEPTED BOOK OF MEDICAL PARASITOLOGY BY K. D. CHATTERJEE
TEXT BOOK OF MEDICAL PARASITOLOGY BY PANIKER
IMAGE SOURCES : FROM INTERNET
LUMEN DWELLING FLAGELLATES - GIARDIA
REFS:
INTERNATIONALLY ACCEPTED BOOK OF MEDICAL PARASITOLOGY BY K. D. CHATTERJEE
TEXT BOOK OF MEDICAL PARASITOLOGY BY PANIKER
IMAGE SOURCES : FROM INTERNET
PowerPoint about laboratory diagnosis of Taenia species (T.saginata and T. solium ) .Contain information about diagnosis with picture to simplify understanding .
Protozoa of this group possess one or more whip like flagella as their organs of locomotion
Classification
According to their habitat
Lumen dwelling flagellates
Alimentary canal – Intestinal flagellates
Urogenital tract – Genital flagellattes
2.Hemoflagellates - flagellates found in blood and tissues
Pathogenic:
Intestinal flagellates - Giardia lamblia Duodenum, Jejunum -Diarrhoea.
Genital flagellates - Trichomonas vaginalis Vagina, Urethra -Vaginitis , Urethritis
Non pathogenic:
Trichomonas tenax ( Mouth)
Trichomonas hominis ( Caecum).
Enteromonas hominis ( Colon)
Dientamoeba fragilis( Colon)
PowerPoint about laboratory diagnosis of Taenia species (T.saginata and T. solium ) .Contain information about diagnosis with picture to simplify understanding .
Protozoa of this group possess one or more whip like flagella as their organs of locomotion
Classification
According to their habitat
Lumen dwelling flagellates
Alimentary canal – Intestinal flagellates
Urogenital tract – Genital flagellattes
2.Hemoflagellates - flagellates found in blood and tissues
Pathogenic:
Intestinal flagellates - Giardia lamblia Duodenum, Jejunum -Diarrhoea.
Genital flagellates - Trichomonas vaginalis Vagina, Urethra -Vaginitis , Urethritis
Non pathogenic:
Trichomonas tenax ( Mouth)
Trichomonas hominis ( Caecum).
Enteromonas hominis ( Colon)
Dientamoeba fragilis( Colon)
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
These lecture slides, by Dr Sidra Arshad, offer a quick overview of 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 leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
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. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Knee anatomy and clinical tests 2024.pdfvimalpl1234
This includes all relevant anatomy and clinical tests compiled from standard textbooks, Campbell,netter etc..It is comprehensive and best suited for orthopaedicians and orthopaedic residents.
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.
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
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
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
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.
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.
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.
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...
Diagnostic procedures of schistosomiasis
1. COLLEGE OF MEDICINE AND HEALTH SCIENCES
SCHOOL OF HEALTH SCIENCES
BLS Dpt
Year 4
PARASITOLOGY IV
ASSIGNMENT
TOPIC: Diagnostic procedures of Schistosomiasis
in high and low endemic areas/immigrants
GROUP 10 MEMBERS
215040646 NSENGIYUMVA Emmanuel
215040593 NSENGIMANA Bernard
215040715 NSENGIYUMVA Prosper
2. INTRODUCTION
• Schistosomiasis is parasitic disease caused by schistosoma species.
affect more than 200 million people worldwide, and has been
classified as the second most common tropical disease following
Malaria.
• The schistosomes are a group of closely related flukes that inhabit the
portal vascular system of a number of animals(blood flukes).
• Species which infect human: S. mansoni,, S. haematobium, and S.
japonicum, S. Intercalatum, and s. mekongi.
• Transmission: cercariae penetration of human host skin
• Schistosomiasis manifestation: Schistosomiasis manifested in three
stages, The first is initiated by the penetration and migration of the
schistosomula, the second or intermediate begins with oviposition and
is associated with a complex of clinical manifestations (fever and
chills, patients experience cough, urticaria, arthralgia,
lymphadenopathy, splenomegaly, abdominal pain, and diarrhea )and
The third or chronic stage is characterized by granuloma formation
and scarring around retained eggs.
3. • katayama syndrome: syndrome characterized by
leukocytosis; marked peripheral eosinophilia; and
elevated levels of IgM, IgG, and IgE
immunoglobulins
• Infective stage: Cercariae
• Definitive host: Human
• Intermediate host: snails, bulinus spp , oncomelania
spp, and biomphalaria spp for schistosoma
haematoboum, s. japonicum,and s. mansoni
respectively
• Reproduction: the schistosomes are dioecious,
sexual dimorphism; the male surrounds the female
and encloses her within his gynacophoric canal for
the entire adult lives of the worms.
• Treatment: Praziquantel (PZQ) is the drug of first
choice for treatment of all types of schistosomiasis
5. DIAGNOSTIC PROCEDURES OF SCHISTOSOMIASIS IN
HIGH AND LOW ENDEMIC AREAS/IMMIGRANTS
• Schistosomiasis diagnostic procedures in low endemic
area use tests or procedures with high sensitity and
specifity than in high endemic area in order to find even
light infections
• Laboratory procedures includes direct methods and
indirect methods.
• Direct methods are used for detection of eggs, while
indirect methods are used for detecting specific
antibodies and antigens.
• Sample : urine for urinary schsitosomiasis (S.
hematobium), blood and/or serum, stool for intestinal
schistosomiasis, biopsy, and lavages
6. MICROSCOPIC EXAMINATION
• For the diagnosis of schistosomiasis,
microscopic detection of excreted eggs in stool
or in urine remains the gold standard.
• In direct examination a small quantity of the
faeces, is placed on glass slide along and drops
of physiological or normal saline are added.
• The faecal sample is thoroughly emulsified
,evenly spread over the slide, covered with
cover slide and examined under the low power
of microscope.
7. Cont’
• This method is easy and useful in heavy
infection.
• In case no egg is found, does not mean human
host is worm free.
• So, Varieties of techniques are available such as;
direct concentration, sedimentation concentration,
concentration floatation and filtration assays to
increase sensitivity.
• The principle of these methods is to concentrate
all the eggs in a given amount of faeces.
8. Direct concentration(Kato katz )
• In the Kato-Katz technique faeces are pressed
through a mesh screen to remove large particles.
• A portion of sieved sample is then transferred to the
hole of a template on a slide. After filling the hole,
the template is removed and the remaining sample
is covered with a piece of cellophane soaked in
glycerol.
• The glycerol clears the faecal material from around
the eggs. The eggs are then counted and the number
calculated per gram of faeces.
9. Sedimentation concentation
• The majority of trematode eggs are too
large and heavy to float reliably in the
flotation fluids normally used for
nematode eggs.
• They do however sink rapidly to the
bottom of a faecal/water suspension and
this is the basis of the faecal
sedimentation technique
10. Principle of Formal ether (Formalin-Ethyl
Aceatate) sedimentation technique
• Sedimentation techniques use solution of
lower specific gravity than the parasitic
organisms, thus concentrating the latter
in the sediment.
• Ethyl acetate is used as an extractor of
debris and fat from the feces and leaves
the parasites at the bottom of the
suspension.
11. Floatation Concentration technique
• Eggs are separated from faecal material and
concentrated by a flotation fluid of an appropriate
specific gravity. The principle is to use an emulsifying
fluid of a greater specific gravity than that of the parasite
eggs, which result in flotation of eggs in the solution.
Thereby mere examination of the solution in the top
most layer will clearly indicate the presence of egg. The
faecal material and fiber settle at the bottom
Filtration assays
• Urine filtration is recommended by the WHO for S.
haematobium. A standard 10-ml of the urine to be tested
is forced through the device with a syringe. If eggs are
present, they are unable to pass through a Millipore filter
and can be observed and counted under the microscope
12. McMaster counting technique
• This is a quantitative technique to determine the
number of eggs present per gram of faeces
(e.p.g.). A flotation fluid is used to separate eggs
from faecal material in a counting chamber
(McMaster) with two compartments.
• Multiply the total of eggs in 2 chamber by 50 .
This gives the e.p.g. of faeces. (Example: 12
eggs seen in chamber 1 and 15 eggs seen in
chamber 2 = (12 + 15) x 50 = 1350 e.p.g.)
14. Diagnostic procedures of Schistosomiasis low
endemic areas/immigrants
• Diagnosis of schistosomiasis in low
endemic area and in immigrants is
done with serological tests and
molecular biology tests to measure
even less infection that can not be
seen on fecal or urine microscopic
exam.
15. Serological tests
• Serological tests done by detecting
Antibody and/or antigen in sample of
infected patients who have traveled in
schistosomiasis endemic areas and in
whom eggs cannot be demonstrated in
fecal or urine specimens .
16. Antibody detection
• This performed with Detection of antibodies to
schistosoma adult worm microsomal antigen
including mansoni adult worm microsomal
antigen [MAMA], haematobium adult worm
microsomal antigen [HAMA], japonicum adult
worm microsomal antigen [JAMA]) using
Falcon assay screening test (FAST), enzyme-
linked immunoassay (ELISA), and immunoblot
assays.
• In acute presentation of katayama syndrome,
Antibody detections are generally negative, even
though serology often becomes positive before
eggs become detectable. Seroconversion
generally occurs 4-8 weeks after infection.
17. Antigen detection
• In infected persons, Schistosome antigens are present in
serum and urine, these antigens are referred as CAA
(circulating anodic antigen) and CCA (circulating
cathodic antigens) according to their migratory behaviour
in immunoelectrophoresis.
• These two circulating adult worm antigens are the basis
of antigen capture immunoassays. Measurement of CAA
in the blood, serum, and urine by ELISA-based assays is
sensitive, specific and much less variable than egg
counts. The CCA assay has been further developed as a
point of care (POC) urine ELISA dipstick.
• These tests can differentiate between active and past
infections, as the circulating antigens are probably
present only when there is active infection
18. Test used in schistosoma antigens and
antibodies detection
Test used are:
• Indirect immunofluorescence (IIF) and
• Enzyme-linked immunosorbent assay (ELISA)
• Circumoral precipitin test (COPT),
• the cercaria-Hullen reaction (CHR), and
• Total and fractionated soluble adult worm antigens
(SWAP),
• Bilharziose Fumouze IHA,
• Schistosoma mansoni IgG-ELISA,
• Schistosoma serology microwell ELISA and
• West blotting for confirmation of ELISA test.
19. Schistosoma ELISA Test Principle:
• This test use microwells that are coated with Schistosoma
recombinant antigen.
• Firstly diluted patient sample is incubated in microwells
and containing antibodies which are reactive to antigen
bind to coated microwells and unbounded antibodies are
washed away by washing step,
• secondly, enzyme conjugate anibodies are added and react
with these antibodies, then unbound conjugate antibodies
are washed away,
• thirdly, substrate is added to react with enzymes
conjugated with antibodies, the substrate color will turn
blue.
• Finally the reaction is ended with the stop solution, and
what appears is a yellow color instead of the blue
(Automation/Cortez & Diagnostics 2015).
20. Schistosoma Indirect Fluorescent-
Antibody Tests(IFA-Test)
• Indirect fluorescent-antibody tests are used to
demonstrate the presence of antibodies against a
Schistosoma specific antigen in serum, urine and
stool.
• Antigen is incubated with the patient's serum,
Excess serum is washed away, leaving only
antibodies specific for the antigen present in the
patient's serum bound.
• Then, antibodies labeled with fluorescent dye that
are specific for human antibodies is added. The
sample is viewed with a fluorescence viewer.
21. Indirect Haemagglutination Test (IHT)
• The IHT detects reactivity between antibodies in
the serum of an infected individual and
schistosome antigen-coated red blood cells
which is indicated by agglutination.
• Due to its simplicity and relatively high
sensitivity, this test has been widely used in
large scale community surveys and for
surveillance tool studies in schistosome-
endemic areas.
22.
23. Circumoval precipitin test (COPT)
• The circumoval precipitin test (COPT) is based on
patient serum precipitation with lyophilized eggs or
purified live eggs identified under microscope.
• This method is useful for the diagnosis of S.
mansoni and S. japonicum.
• One drop, about 0.025 mI, of the suspension
containing living larvae, eggs or 6-8 specimens of
immature adults is put into the well of a slide, and 3
drops, about 0.075 ml, of serum is added.
• A cover slip is placed over the well. The slide is
incubated at 34 DC. After 24 hr, the slide is
examined by microscope for the appearance of
precipitates attached to the worms or eggs.
25. Cercarian hullen reaction(CHR)
• The test is simple rapid and sensitive.
• A positive reaction is indicated by formation
of an envelope or a preceracrial sheath around
the ceracariae when incubated in the positive
sera.
26. Fractionated soluble adult worm antigens
(SWAP)
• Soluble adult worm antigens (SWAP) of
Schistosoma mansoni are fractionated by fast
protein liquid chromatography (FPLC) system,
using Q-Sepharose anion-exchange resin, in order to
characterize antigenic fractions that may elicit cell
responses in human schistosomiasis.
• SWAP fractions eluted. The FPLC system resolves
6 fractions, enumerated I to VI, according to the
NaCl gradient.
• The analysis of each fraction on SDS-PAGE showed
that fractions I to IV are constituted by multiple
protein bands with M, ranging from 21 to > 200
kDa.
27. • Large amounts of nucleic acids evidenced in
fractions V and VI, as revealed by ethidium
bromide staining of agarose electrophoresis
gels. Using ELISA, it shows that sera from
chronic schistosomiasis patients contained
antibodies that recognized antigens in
practically all fractions.
28. Western blot
• Western blot :schistosomula antigens preparation
(SAP) and adult worm antigens proteins are
separated denaturing polyacrylamide gel by
electrophoresis according to size, charges and
molecular weight. From the gel, the proteins are
transferred to nitrocellulose membranes.
• The nitrocellulose membranes is blocked with 5% dry
milk
• Appy primary antibody against these proteins.
• After three washes, the membranes are incubated anti-
mouse IgG-alkaline phosphatase conjugate.
• After three washes, the membranes are treated with
alkaline phosphatase reaction (Carvalho et al. 2011) .
29. Western blot cont’
• Results are measured looking for bands
with colors.
• Ex. Recognition of proteins with
approximately 200 kDa, 100 kDa, 43 kDa
and 18 kDa by sera from infected mice in
AWP similar recognition pattern of proteins
from SAP with approximately molecular
weight of 100 kDa, 43 kDa and 18 kDa by
sera of infected mice was observed for
diagnosis of S.mansoni (Carvalho et al.
2011).
32. Reagent strip test
Reagent strip for antigens
• Reagent strip used by detecting parasite antigen in urine
of infected individuals. The principle of test is based
lateral flow through a nitrocellulose strip of the sample
mixed with a colloidal carbon conjugate of a monoclonal
antibody specific for Schistosoma circulating cathodic
antigen (CCA).
• The presence of the analyte (CCA) is made visible by
capture of the immune complex of antigen and
carbon-labeled antibody by the anti-CCA monoclonal
antibody that is immobilized on the strip as a test line.
• In addition, a line of immobilized polyclonal anti-mouse
antibodies is used to capture the excess carbon-labeled
antibodies to act as a positive control line.
34. Point of care (POC) – CCA dipstick.
• The urine POC- CCA test is a lateral flow
immuno-chromatographic urine dipstick assay
that uses a nitrocellulose strip with a
monoclonal antibody coated test line to detect
the presence of Schistosoma-specific CCA
antigen in urine.
• When urine from an infected individual flows
through the strip, the antigen will bind to the
test line, which becomes visible with the
binding of added labelled monoclonal
antibodies.
36. Reagent strip test for blood in urine
(heamaturia)
• This test is based on the pseudoperoxidase
action of hemoglobin and erythrocytes which
catalyzes the reaction of 3, 3’, 5, 5’-
tetramethyl-benzidine and buffered organic
peroxide.
• The resulting colors range from orange to
yellow-green and dark green. Very high blood
concentration may cause the color
development to continue to dark blue
(Diagnostics 2008).
37.
38. Intradermal test
• The test is made intradermally injection of antigen
from adult worm, cercaria and snail, the aim being
to inject 0.01 cc. only. It is read in 10, 15 and 20
minutes, and a positive result is indicated by a
raised, button-like weal, which may have
pseudopodia.
• Intradermal test has widely been used in screening
human population for presence of schistosomiasis
in endemic countries with good results, both
immediate and delayed reactions are recorded
after 15 min and 24 hour of application
respectively
39.
40. Molecular biology tests
• Molecular approaches can be used to detect DNA of
different life cycle stages of S. mansoni by analyzing
different biological samples: feces, snail tissues and
infested water bodies, resulting in diagnosis of vertebrate
and invertebrate host infections and identification of
transmission sites
• The application of PCR as a technique for the detection
of schistosomiasis has been explored for S. mansoni and
S. japonicum in human faeces and urine in areas of
medium and low intensity of infection.
• PCR
• DNA for amplification can be extracted from adult
worm, eggs, and cercaria stages, these can be ribosomal,
mitochondria and nuclear DNA.
41. PCR Amplification
Materials: PCR primers (forward and reverse) ,The reaction
volume of 25 μL consisted of 1.25 units Taq DNA polymerase,
2.5 μL 10× buffer, 1.5 mM MgCl2, 200 μM (each) of dATP,
dCTP, dGTP, and dTTP, 1 μM of each of the amplification
primers and 5 μL (3 ng) of template DNA.
Steps of PCR
• Activation /denaturing step of 15 min at 95°C
• Annealing temperature of 53°C for 1.5 min, and expansion at
72°C for 1 min
• Final extension step at 60°C for 5 min.
• Amplification is conducted in 0.2 mL PCR tubes in a thermal
cycler. The products are analyzed on a 2% agarose gel stained
with ethidium bromide (10 mg/μL) and visualized with UV
light. Bp ladder are used to estimate band size (Ibironke et al.
2011).
• Types of PCR used are real PCR, nested PCR,
44. Loop-mediated isothermal amplification
(LAMP)
• Loop-mediated isothermal amplification
(LAMP) is a unique amplification method with
extremely high specificity and sensitivity able
to discriminate between a single nucleotide
difference.
• It is characterized by the use of six different
primers specifically designed to recognize
eight distinct regions on a target gene, with
amplification only occurring if all primers bind
and form a product
45. Histological tests
• Biopsy is helpful when stool sample findings are
negative or in light infection.
• Mucosal biopsy is effective for visualizing eggs.
• Rectal or bladder biopsy.
• Sigmoidoscopy/proctoscopy: to obtain mucosal
biopsies for diagnosis and to identify
complications such as pedunculated and sessile
polyps
• Cystoscopy : is useful in schistosomiasis with
primary bladder involvement for ulcers, polyps ,
hematuria, dysuria.
46. histology
• Surgical biopsy: used to identify ectopic
schistosomiasis
• Bronchoscopic washings or transbronchial
biopsies : for determining Eggs individuals with
pulmonary involvement.
• Lumbar puncture : for Eosinophils present in
the cerebrospinal fluid (CSF) of individuals with
neurologic involvement.
• Upper endoscopy : to assess for esophageal
varices; treat upper intestinal bleeding with
endoscopic sclerotherapy
47. AA, S Mansoni, B: S. mansoni, C: S. Japonicum, D: S. hematobium in urinary bladder
biopsy
48. Adults worms(cross sectional cuts)
Adults of Schistosoma spp. in lung tissue, stained with
H&E. Images courtesy of Harvard Medical School,
Cambridge
49. Imaging tests
• Ultrasonography: for assessing hepatosplenic disease
with periportal fibrosis or urinary obstruction. It can
demonstrate periportal fibrosis, splenomegaly, portal
collaterals, periportal adenopathy, ureteral obstruction,
and obstructive nephropathy.
• Echocardiography and/or invasive hemodynamic
studies can demonstrate pulmonary hypertension and
cor pulmonale, if present.
• Chest radiographs may show patchy infiltrates in
acute schistosomiasis and can indicate pulmonary
hypertension and cor pulmonale in end-stage chronic
infection, if present.
• Computed tomography (CT) or magnetic resonance
imaging (MRI) scanning may be useful in the
evaluation of CNS disease or in the detection of
periportal fibrosis.
50. A.calcified bladder in a chronic form of schistosomiasis, B. Liver
fibrosis in hepatosplenic schistosomiasis (white areas around the
vessels). There is a thrombus inside the portal vein (red arrow)
A B
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