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Pathology
Jaroslava Dušková
Inst. Pathol. ,1st Med. Faculty,
Charles Univ., Prague
Bridging
Medical
Branches
General Pathology I
Introduction
– where to find information
– who is a pathologist
– what is pathology
health and disease
Activities in pathology
– autopsy
– biopsy
– cyto(patho)logyPROGNOSTICATION-PREDICTION- PREVENTION
Methods
– macroscopy
– light microscopy
– ultrastructure (electron microscopy)
– molecular pathologyProf. Jaroslava Duskova, MD, PhD, FIAC
http://pau.lf1.cuni.cz/
Pathologist !?!
Pathologist ????
Pathologist =anonymous diagnostic service
for ALL medical specialisations
Pathologyscience on diseases
– causes (etiology)
– mechanisms of
formation (pathogenesis)
– development (prognosis)
Disease
– health disturbance
Health (WHO) – status of full
physical and mental and social
wellfare and harmony
Disease – adaptation mechanisms overcome
Disease - manifestation
features– symptoms
– general - local
– subjective - objective
sets of features – syndromes
W-F sy
Diagnosis – disease identification
Based on
history
symptoms
targeted search in asymptomatic
(screening)
laboratory tests and imaging
Differencial diagnosis !!!
Disease - description
Etiology – cause(s)
Pathogenesis – development
Complications
Healing (sequela/e)
Prognosis
Causes of
diseases
External
– physical
– chemical
– biological
– environmental
– nutritional
Internal
– genetic
– disposition
(tendency)
– immune disorders
hypersensitivity
(alergy)
immunodeficiency
autoimmunity
Pathology general
special– organ
– nosological(nosos= illness)
Dissection in the University of
Bologna in the 14th century. The
body, of an executed criminal, is
being studied by a group of
students.
New anatomical theatre,
University of Rome, Italy,
1785-1805.
Steps from the History of Pathology
(starting with the „anatomical“ era)
Andreas Vesalius (1514-1564) 1543
Jan Jesenius (1566-1621 1600
G.B.Morgagni 18th cent.
A.van Leuwenhoek
Steps from the History
of Pathology 19th cent.
R.L.K. Virchow
I.P.Pavlov
K. Rokitansky
V.Treitz
V.D.Lambl
German anatomist and pathologist,
• Rudolf Ludwig Karl Virchow (1821-
1902). studied medicine at Berlin
University (1840-1843)
• working as a surgeon in the Charite
Hospital in Berlin
• lectures on pathological and surgical
anatomy.
• In 1847 he established the Archives for
Pathological Anatomy, Physiology &
Clinical Medicine,
• In 1855-1856 he published his major
work 'Collected Essays for Scientific
Medicine', which was followed in 1859
by his 'Treatise on Constitutional
Syphilis'.
Karl Freiherr von
Rokitansky (1804-1878)
Austrian pathologist,
born February 19, 1804, Königgrätz, Böhmen, Austrian Empire (now Hradec Králové, East
Bohemia, Czech Republic);
died July 23, 1878, Wien.
Handbuch der
pathologischen
Anatomie IInd Band,
Wien 1842
The Vienna School of Pathology
(& Czech Specialists)
Karl Rokitansky, Josef Škoda-signatures from the memorial book
1837 - first autopsy (protocol preserved in the
institute archives)
pathology -
compulsory subject
founder of the
MUSEUM
VINCENC ALEXANDER
BOCHDALEK
1858
VÁCLAV
TREITZ 1819-1872 Hernia retroperitonealis -
Treitzi
Herwig
Hamperl
- 1945
Chairman of the German Institute of
Pathology, Charles University, Prague
in years 1940-1945.
Werdegang und Lebensweg eines
Pathologen . Stuttgart Vrlg. 1972
A Pathologist's Life and Evolution
Steps from the History
of Pathology 20th cent.
Jaroslav Hlava
Heřman Šikl
Antonín Fingerland
Blahoslav Bednář
Ctibor Povýšil
editors of Czech textbooks
of pathology
Activities in Pathology
Observation subject
necropsy (autopsy)
biopsy
cytology
experiment– animals– tissue cultures
molecular techniques
Methodology of Pathology
Observation
level
macroscopy
microscopy
ultrastructure
Special methods
histochemistry
immunohistochemistry
quantitative (computer)
image analysis
genetic methods
– FISH, PCR, CGH, …
AutopsyAfter the 2nd world war standing decrease
abroad < 10%, Czech Rep. < 50%
Aims– clinicopathological diagnosis
– cause of death
– dg. errors
– therapy effects assessment
– data on old and new diseases
– teaching
– event. medicolegal aspects of death
Autopsy
dissection method unchanged
since the end of 19th century
changes in taking material for
special methods & their
employment to cope with clinical
questions
Ulcus chronicum pepticum pylori
Helicobacter pylori (Warthin – Stary impregnation)
Clinicopathological Diagnosis
Morbus principalis
Complicationes
Causa mortis
Inventus accesorius
Epicrisis
Biopsy
After the 2nd world war standing increase
Aims
– nosological diagnosis
– tumour pathology
– typing, staging, grading
– evaluation of therapy efficiency
Biopsy sample - fixed
Basic staining
H&E
van Gieson
and elastics
Atherosclerosis gr. II. a. coronariae cordis
Myofibrosis ventriculi sin. cordis
alc. blue, pH 2,5
Carcinoma mammae lobullare invasivum alc. blue pH2,5
signet ring cells
Fe
Klűver- Barrera
Immunohistochemistry
detection of an antigenic component in
the histology section
helpful in identifying cellular &
extracellular components
part of daily routine & investigation
useful but limited (like any other method)
Immunohistochemistry - steps
blockage of non-specific positivities- esp.
endogenous peroxidase
incubation with a commercially available
specific antibody
visualisation of the positive reaction
Direct immunohistochemical method
Primary antibody
Antigen
Peroxidase
Primary antibody
Secondary antibody
Streptavidin-biotin method. Enzyme marked
avidin or avidin-biotin complex with enzyme
marked biotin reacts with biotinylated secondary
antibody.
Peroxidase
Leiomyoma intestini crassi (MSA immunohistochemistry)
H&E Fe
TGB
PTH
Adenoma glandulae
parathyreoideae
Carcinoma mammae lobulare invasivum ILC
HE ER
PRalc. blue 2,5
Carcinoma mammae ductale invasivum IDC
HE PRER
MIB1 e-cadherin c-erbB2 / Her2-neu
„Classical“ Biopsy (formol paraffin technique)
simple may be done in two days
immuno reactions two days more
further sectioning two days more
oncol. dg. - WHO classification
Peroperation Biopsy-Frozen section
reported DURING MINUTES !!!
morphological artefacts
(combination with cytology)
limited extensity of investigation
limited time
Cytology
After the 2nd world war standing increase
Aims
– screening
– nosological diagnosis
– evaluation of therapy efficiency
Diagnostic Cytopathology
Jaroslava Dušková
Inst. Pathol. ,1st Med. Faculty, Charles Univ. Prague
Gynecological
oncologic
cytology
laboratory investigation standard
Authors: MUDr Alena Beková, MIACMUDr Pavel Tretiník, MIAC
Oponents: doc. MUDr J. Dušková, CSc,FIACMUDr Eva Svobodová
SIL H
invasive ca
F
N
A
B
respiratory
epithelia
muscle
fatty
tissue
Morphological Diagnostic Methods
Clinical
Pathological
Morphological Diagnostic Methods
Clinical:
macroscopy of lesions
visible with the naked eye
invisible with the naked eye -
IMAGING (X-ray,
sonography, scintigraphy, endoscopy,
CT,…)
magnifying glass - colposcopy
Morphological Diagnostic Methods
Pathological
macroscopy
microscopy
ultrastructure
IMAGING
Archiving- sending- wireless consultations…..
Morphometric Investigations
Objectivisation & refinement
of diagnostic methods
staging (e.g. malignant melanoma –
thickness of the neoplasm)
grading (e.g. kidney carcinoma – size of the
nuclei)
Carcinoma intestini crassi
Malignant Melanoma measurement - a compulsory part of biopsy report
Breslow – max. thickness [mm]
Clark – level of skin involvement
AgNOR Measurements
Morphometric evaluation of
the Focus Score
in the LUCIA system
Jaroslava DuškováLadislav Korábek
Inst Pathol. and Stomatology Clinic 1.st Med. Faculty, Prague
Focus Score
morphometric expression of intensity of
focaly accented chronic sialoadenitis
focus – agregate of 50 and more
lymphocytes(defined 1968)
Ultra-
structural
morphometry
of internal
limiting
membrane
of retina
Dg. H35.3 Degeneration of
macule and posterior eye pole
Epimacular
membrane of retina
Ultrathin section
5000x
Semithin section 400x
Molecular Genetic Methods
PCR, FISH, CGH, Hybrid capture, New
Generation Sequencing…
become a part of daily rourine diagnostics of
neoplasms and other diseases
– prognostic, and
– predictive (therapeutic response) – EGFR,
VEGF, k- ras….
histopathological & cytopathological samples
suitable
Molecular Genetic Methods
PCR, FISH, CGH, Hybrid capture, Next-
Generation Sequencing…
become a part of daily rourine diagnostics of
neoplasms and other diseases
– prognostic, and
– predictive (therapeutic response) – EGFR,
VEGF, k- ras….
histopathological & cytopathological samples
suitable
Invasive ductal breast cancer with signal amplification
c-erbB2 (Her2-neu)
Hybrid capture system– DML 2000 microtitration plate– 8channel pipette– disposable pipettes– heater block– rotary shaker – luminometer– pc– sw– printer
Detection of LR & HR
HPV1. specimen denaturation
2. hybridization with HPV RNA-
probes
3. capture hybrids RNA/DNA using
monoclonal AB on microtiter plates
4. second monoAB marked with
alkaline phosphatase AP
5. AP splits chemoluminiscent
substrat
QUANTIFIABLE RESULT
detection of 5 types LR and 13 types HR HPV
Particitation of Pathologists
in the Czech National Screening Programs:
Breast cancer: mammography – core biopsy- resecate
Colon Cancer: occult bleeding test– endoscopy – endoscopic
biopsy - resecate
Cervical cancer: Pap test – colposcopy - punch biopsy - resecate
Pathology an extremely exciting medical
specialty
helping to understand & treat the diseases
via
diagnostic service
for ALL other specialties