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