7/21/2019 2007 Malaria

1/108

7/21/2019 2007 Malaria

2/108

-- 1 -

3 23-24 2550

PCR

9 2550

7/21/2019 2007 Malaria

3/108

-- 2 -

1 3 5 8 11 18 26 31 39PCR : 86 95 99

100

103

7/21/2019 2007 Malaria

4/108

-- 3 -

Diagnosis of Malaria as a Professional

3 23 24 2550

-----------------------------------------------------

23 255008.30 09.00 . 09.00 10.30 .

10.30 10.45 . 10.45 - 12.00 .

12.00 13.00 . 13.00 14.00 .

14.00 14.45 .

14.45 15.00 . 15.00 15.45

.

15.45 16.30 .

7/21/2019 2007 Malaria

5/108

-- 4 -

24 255008.30 09.00 .

09.00 10.00 .

10.00 10.15 . 10.15 12.00 . Thick and thin smear

12.00 13.00 . 13.00 14.45 . WHO

14.45 15.00 . 15.00 16.00 . PCR :

16.00 16.30 .

-----------------------------------------

7/21/2019 2007 Malaria

6/108

-- 5 -

1.

.. 2518 300

1-2

7/21/2019 2007 Malaria

7/108

-- 6 -

/

2. 2.1 2.2 2.3

PCR antigen detection2.4

3. 23 24 2550 3

4. /

5.

- - - -

7/21/2019 2007 Malaria

8/108

-- 7 -

- - Thick and thin smear - WHO

-

PCR :- Rapid Diagnostic test Kit

6. 6.1 /

6.2 / PCR antigen detection

6.3/

6.4/

6.5

6.6

7.

. 0-2354-9100 1830 1833 0-2643-5601

7/21/2019 2007 Malaria

9/108

-- 8 -

1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 12.

1. 2. 3. 4.

5. 6. 7.

7/21/2019 2007 Malaria

10/108

-- 9 -

1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11.

1. 2. 3. 4.

1. 2.

3.

7/21/2019 2007 Malaria

11/108

-- 10 -

1.

2. 3. 4. 5.

1. 2. 3. 4. 5.

________________________

7/21/2019 2007 Malaria

12/108

-- 11 -

.. 1, . 2, . 3,.. 2, .. 2

12

3

Genus Plasmodium

1600 (1)

- -

10-35 2

.. 2549 30,338 161 65 2

7/21/2019 2007 Malaria

13/108

-- 12 -

- -

(1) (recrudescence) 2 (hypnozoite)

(relapse) hypnozoite

primaquine primaquine primaquine (redical cure)

3-6

1

7/21/2019 2007 Malaria

14/108

-- 13 -

(influenza) (denguehemorrhagic fever) (viral hepatitis)

(fulminant hepatitis withhepatic encephalopathy)

(bacterial sepsis) (pneumoniawith pulmonary edema) (feverof unknown origin)

2-6

7

( 15%

5 g/dl)

/DIC (2 2 ) (pH 7.35 bicarbonate 15

mmol/l base excess 10) (macro hemoglobinuria) Prostration

7/21/2019 2007 Malaria

15/108

-- 14 -

hyperparasitemia

1.

2. (antigen)

(rapid diagnostic test) Optimal test,Paracheck, ICT test

3. (antibody)

4.

(DNA RNA)

5. Quantitative buffy coat (QBC)analysis

8 2

1

7/21/2019 2007 Malaria

16/108

-- 15 -

1 dipsticks

HRP-2 pLDHpLDH HRP-2

False positive < 5

False positive < 7-21

15 (OptiMAL) 3-5 (OptiMAL)

1 1 Asexual & young

gametocyte

+++ (OptiMAL) ++ (ICT)P.falciparum

+++ (OptiMAL) ++++ (ICT)

++++ (OptiMAL) +++ (ICT)false positive ? Rheumatoid factor ? Rheumatoid factor 70-90 100 Certificate of free sell Certificate of free sell NaN3 NaN3

7/21/2019 2007 Malaria

17/108

-- 16 -

blood smear influenza, viral hepatitis, dengue

hemorrhagic fever, pneumonia, leptospirosis

intensive care respirator

dialysis

corticosteroidacute respiratory distress syndrome (ARDS) dialysis

platelet concentrate systemicbleeding

7/21/2019 2007 Malaria

18/108

-- 17 -

1. Breasted JH. "The Edwin Smith Surgical Papyrus." Chicago, Illinois:

University of Chicago Press, 1930.

2. [Online]. 2549 [cited 2007 Jan]; Available from: URL:http://www.thaivbd.org/cms/index.php?option=com_content&task=view&id=57&I

temid=1

3. Bunnag D, Karbwang J, Thanavibul A, Chittamas S, Rattanapongse Y,

Chalermrut K, et al. High dose of primaquine in primaquine resistant vivax

malaria. Trans R Soc Trop Med Hyg 1994;88(2):218-219.

4. Looareesuwan S, Buchachart K, Wilairatana P, Chalermrut K, Rattanapongse

Y, Amradee S, et al. Primaquine-tolerant vivax malaria in Thailand. Ann Trop

Med Parasitol 1997;91(8):938-943.

5. Wilairatana P, Silachamroon U, Krudsood S, Singhasivanon P, Treeprasertsuk

S, Bussaratid V, et al. Am J Trop Med Hyg 1999;93(6):973-977.

6. Silachumroon U, Krudsood S, Treeprasertsuk S, Wilairatana P, Chalermrut K,

Mint HY, et al. Am J Trop Med Hyg 2003;69(1):14-18.

7. Severe falciparum malaria. World Health Organization, Communicable

Diseases Cluster. Trans R Soc Trop Med Hyg 2000;94 Suppl 1:S1-90.

8. The use of malaria rapid diagnostic tests. Geneva: World Health Organization,

2004. pp 19.

----------------------------------------------

7/21/2019 2007 Malaria

19/108

-- 18 -

.

.. 1955 300

1-2

(Plasmodium)

4 (species)

(Plasmodium falciparum) (Plasmodium vivax)(Plasmodium malaria (Plasmodium ovale)

(P.falciparum)

7/21/2019 2007 Malaria

20/108

7/21/2019 2007 Malaria

21/108

-- 20 -

(Schizogony)

sporozoite merozoitemerozoites merozoites 4 - 36

merozoite merozoite (gametocyte)

Plasmodium falciparum

Plasmodium malaraie merozoite merozoites Plasmodium vivax

Plasmodium ovale 2 merozoite Hypnozoite

(reactivate) merozoite relapse

(Sporogony)

gametocytes gametes zygote ookinete oocyst sporozoite

oocyst sporozoite (salivary gland) (salivary duct) sporozoite

(Morphology)

(species)

7/21/2019 2007 Malaria

22/108

-- 21 -

1. Early trophozoite Ring form chromatin 1-2 cytoplasmvacuole

2. Growing trophozoitecytoplasm pigment

3. Late trophozoite vacuole chromatin pigment

4. Early schizont chromatin 1 2

5. Growing schizont chromatin 2 cytoplasm

6. Mature schizont chromatin cytoplasm merozoite

pigment merozoites

7. Gametocyte cytoplasm vacuole chromatin cytoplasm pigment

cytoplasm dots

Giemsa

7/21/2019 2007 Malaria

23/108

-- 22 -

P. vivax P. falciparum P. malariae P. ovale

EE.schizogony 8 6 9 7-12 E.schizogony 48 48 72 48

1.4

1.2

Stipping Schuffners

dot

Maurers dot Ziemanns dot Schuffners dot

Reticulocyte

(86%)

Reticulocyte &

erytrocyte

Erytrocyte reticulocyte

/

trophozoite

schizont

gametocyte

ring

gametocyte

Trophozoite

schizont

gametocyte

trophozoite

schizont

gametocyte

7/21/2019 2007 Malaria

24/108

-- 23 -

()

P.vivax P.falciparum P.malariae P.ovale

Early

trophozoite

2.5 cytoplasm

chromatin 1

1.251.5

cytoplasm chromatin 1-2

(accole)

P.vivaxCytoplasm

P.vivax

Growing

trophozoite

cytoplasm vacuolepigment

amoeboid

form

cytoplasm

compact pigment

Cytoplasm band form

compactform

cytoplasm vacuole

pigment

Late

trophozoite

cytoplasm vacuolechromatin

growing

trophozoite vacuole

growing

trophozoite cytoplasm

P.vivax

Early

schizont

chromatin

2

chromatin

2

P.vivax P.vivax

7/21/2019 2007 Malaria

25/108

-- 24 -

()

P.vivax P.falciparum P.malariae P.ovale

Growing

schizont

chromatin 2 cytoplasm

chromatin

2 cytoplasm

P.vivax P.vivax

Mature

schizont

merozoite12-24

pigment

merozoite8-36

pigment

merozoite6-12

pigment

merozoite4-18 (8)

Gametocyte

vacuole

cytoplasm pigment

P.vivax P.vivax

Malarial

pigment

1-2

P.vivax

7/21/2019 2007 Malaria

26/108

-- 25 -

Giemsa

7/21/2019 2007 Malaria

27/108

-- 26 -

4

1.

smear 3-5

30-45

2. 2

Romanowskystain

methylene blue eosin eosinatedmethylene blue oxidizedmethylene blue (Azure)

Romanowsky Wright stain, Field stain, Giemsa stain

Giemsa Giemsa 7.0 - 7.2 (pH 7.0 - 7.2)(Phosphate buffer water)

7/21/2019 2007 Malaria

28/108

-- 27 -

2 1. (thick film)

1.1 Field (Fields stain)

A (stain A) 5 5 B (stain B) 3 5

1.2 Modified Wright (Modified Wrights stain)

Wright 1 3

3-5

1.3Giemsa (Giemsas stain) Giemsa

3% coplin jar Giemsa 3% jar 30

7/21/2019 2007 Malaria

29/108

-- 28 -

2. (thin film)2.1 Wright (Wrights stain)

Wright 1 -2

3

2.2 Modified Field (Modified Fields stain)

absolute methanol alcohol 30

B (stain B) 3 5 A (stain A) 5 5

2.3 Giemsa (Giemsas stain)

Giemsa 3%

absolute methanol alcohol 30

coplin jar Giemsa 3%jar 30

7/21/2019 2007 Malaria

30/108

-- 29 -

1. Giemsa

oxidation2. Giemsa

3. Giemsa

deionized water

4. Giemsa

(6 ) (objective lens) 100x

7/21/2019 2007 Malaria

31/108

-- 30 -

1. 2. 3. 4

1. 2. 303. 4.

o Immuno diagnosis Genetic probes (DNA, RNA)

Dye-conjugated MAB-based detection (Dipsticks)

Dot blot

Two-side sandwich ELISA

Immunoradiometric assay (IRMA)

oGene Amplification technique (PCR) for antigen detection

oBiochemical technique

7/21/2019 2007 Malaria

32/108

-- 31 -

.

1. (Microscopy)1.1 (Stained smears) Giemsa Wright's stain

thick thinsmears

1.2

acridine orange

test kit

1.3 Immunofluorescence

microscopy antibody

7/21/2019 2007 Malaria

33/108

-- 32 -

1. thin smear

2. thin smear

7/21/2019 2007 Malaria

34/108

-- 33 -

3. thin smear

4. thin smear

7/21/2019 2007 Malaria

35/108

-- 34 -

2. (Immunoassays)

2.1 histidine rich protein-2 (HRP-2) P. falciparum(5)

2.2. lactate dehydrogenase (pLDH) ezymatic activityimmunoassay (6)

test kit

3. (Molecular biology method)

Polymerase chainreaction (PCR) species

7/21/2019 2007 Malaria

36/108

-- 35 -

5. Amrad ICT Pf immunochromatographic test P. falciparumHRP-2.

6. OptiMAL test pLDH

7/21/2019 2007 Malaria

37/108

-- 36 -

7. 4 PCRLane 1: P. vivax(120 bp)

Lane 2: P. malariae(144 bp)

Lane 3: P. falciparum(205 bp)

Lane 4: P. ovale( 800 bp)

PCR

run gelelectrophoresis Real-time PCR

primers probe 2

primers (fluorescent group) probe 1 probe 2

7/21/2019 2007 Malaria

38/108

-- 37 -

probe 1 probe 2 FluorescenceResonance Energy Transfer

(FRET) ( 8 ) melting curve analysis

4 species probes primers peak meltingtemperature

8. Fluorescence Resonance Energy Transfer

7/21/2019 2007 Malaria

39/108

-- 38 -

References

1. Bain, B.J. et. al. The laboratory diagnosis of malaria. Clin. Lab. Haematol.

1997;19:165-170.

2. Hanscheid, T. et. al. Diagnosis of malaria:review of alternatives to conventional

microscopy. Clin. Lab. Haematol.1999;21:235-245.

3. Tham, J.M. et al. Detection and species determination of malaria parasites by

PCR: comparison with microscopy and with Parasight-F and ICT malaria Pf

tests in a clinical environment. J. Clin. Microbiol. 1999;37:1269-1273.

4. Siribal S, Nakasiri S, Looareesuwan S, Chavalitshewinkoon-Petmitr P.

Identification of human malaria parasites and detection of mixed infection in Thai

patients by nested PCR. Southeast Asian J Trop Med Public Health

2004;35:5-9.

5. Rosenblatt J, Sloan L, Swan H, Chavalitshewinkoon-Petmitr P, Looareesuwan

S, Muyombwe A, Krudsood S, Leowattana W, Wilairatana P. Evaluation of a

real-time PCR assay for the diagnosis of malaria in patients from Thailand.

American Journal for Tropical Medicine and Hygiene 2005; 73:850-854.

---------------------------------------------------------------

7/21/2019 2007 Malaria

40/108

-- 39 -

Class Sporozoa, GenusPlasmodium

1600(1 ) Balanites 400 Hippocrates (2)

Celsus .. 800 -.. 1300

Columbus

(3,4,5)

64o32o 400 Dead Sea 2,600 100

.. 2544 300 1

7/21/2019 2007 Malaria

41/108

-- 40 -

700,000(6)

DuffyNegative

10 (7)

.. 2544 67,749 625

(8)

P l a s m o d i u m (sporozoite)

7/21/2019 2007 Malaria

42/108

-- 41 -

2 2

( ) zygote zygote

oocyst () 2(9)

4

(recrudescence) 2

(hypnozoite) (relapse)

(9 )

hypnozoite primaquine primaquine

(radical cure) primaquine(10-14)0.5

50:50(7)

7/21/2019 2007 Malaria

43/108

-- 42 -

410 72 (15)

1. A n o p h e l e s dirus

2. Anopheles minimus

3. A n o p h e l e s m a c u l a t u s

4. Anopheles sundaicus

5. Anopheles aconitus

7/21/2019 2007 Malaria

44/108

-- 43 -

3

1. 15-60

2.

2-6

3.

3

endogenous pyrogen (interleukin-1) (16)

40 C

(17-19) asymptomatic parasitemia

7/21/2019 2007 Malaria

45/108

-- 44 -

G6PD capsule

post cere bralpsychosis

(20)cellebellar signs(21)

4 malignant malaria

(22 ) ( m e t a b o l i cacidosis)

(23)

4-5

48 tertian malaria

be nigntertain malaria (relapse) hypnozoite

7/21/2019 2007 Malaria

46/108

-- 45 -

(24)

P . v i v a xmultinucleatum

(25)

2 8

(relapse) 3-6 (subtropics) 9

P. v ivax h ibernans 8-9 sporozoite

hypnozoite

1 5

3 13 4

quartan malaria 53

n e p h r o t i csyndrome

7/21/2019 2007 Malaria

47/108

-- 46 -

(mixed infections)2

endemic area

0.5 2 33 (26)2

(27)

3 1.

2Glasgow 8 (30-31)

2.

(asexual form)

3.

7/21/2019 2007 Malaria

48/108

-- 47 -

50 grand mal epilepsy aspiration pneumonia

algid malaria

Cheyne-Stokes breathing metabolicacidosis hypoglycemia

39C

40C

per iphera l

vasoconstriction pulse pressurehigh cardiac output systemic vascular resistance

50% 1-2 .

30%

9% corticosteriod heparin

mild meningealirritation (meningism) board like rigidity

7/21/2019 2007 Malaria

49/108

-- 48 -

lymphocyte5 1 ..cranial nerves

15% nystagmus eye ground retinal haemorrhage Rothsspot flame shape 18% (33)pupillary,oculocepalic, oculovestibular, corneal reflex jaw

jerk 30% primitive reflexpoutreflex finger reflex 5% ankle clonusmuscle tone hypotonus hypertonus

air way endotrachealtube s p a s t i c r i g i d i t y decerebrate rigidity abdominal cremasteric reflex

conversion reaction plantar reflex extensor response

50% 25%

2-3

coma 2 3

5% postmalarial psychosis, post malarial

tremor, cranial nerve 6 hypoglycemia

7/21/2019 2007 Malaria

50/108

-- 49 -

(34)graymatter malaria pigment whitem a t t e r (petechial haemorrhage) (schizont) (capillary) sequestration endothelium cytoadherence

70%(35)

endothelium(knob)

cytoadherance

(36)fibrin

(sequestration) demyelination Durcksgranulomata

(37)

7/21/2019 2007 Malaria

51/108

-- 50 -

sequestration 50-90%

(38)

humanendothe l ia l ce l l amelano t icmelanoma cell

(39)

strain

(40)

knob(41)

knob cytoadherence knob histidine

histidine knob adhesin (42)adhesin 250-300 k endothelial l igand adhesin endothelial ligand thrombospondin(43)adhesinknob cytoadherence endothelial ligand thrombospondin glycoprotein 420 k

ex-vivo (44 )() knobcytoadherance

knob

7/21/2019 2007 Malaria

52/108

-- 51 -

(45)(strain-specificantibody) ame la no t i cme lanoma ce l l a n t i -thrombospondin, monoclonal polyclonal antibody melanoma cell(46)

monoclonal antibody OKM5

cytoadherance(47)Cohen .. 1961

(poo led hyper immuneimmunoglobulin) (48)

anaerobic pathway lactate lactic acidosis lactate CSF (49)

(cerebral blood flow)

(50) high flow

luxury perfusion syndrome

7/21/2019 2007 Malaria

53/108

-- 52 -

(neurological deficit)

hypoxia

1. (rigidity) (viscosity) (51-52)

dextran reticuloendothelial system(53)

(capillary)(54)

venule

2. (agglutinationand sludging) Knisely .. 1941 (55-56)

sludge

endogenous pyrogen interleukin 1 (57 -58 )

interleukin 1 prostaglandinE 2 interferon

(59)monokine tumor

n e c r o s i s f a c t o r ( TNF )( 6 0 )

cachectin TNF interleukin 1(61)

neutrophil TNF procoagulant activity

7/21/2019 2007 Malaria

54/108

-- 53 -

10-50% (27)

coma 1 10% coma 2 20% coma3 4

3 50%

1. 100,000

.. 2 2.

3. coma coma 2-4,

APACHAE SCORE (62)4.

5. bacteria

6. r e t i n a lhaemorrhage

(33)

177 20% 4 bacteria metabolic acidosis (27)

80% 48

2

80%

7/21/2019 2007 Malaria

55/108

-- 54 -

(27)

.. 1946 Spitz (34 )

(14 15)

hyaline membrane monocyte, neutrophil lymphocytemacrophage

(37)

10

1.(fluid

overload) (63)

uremia

2.

Mac-Pherson (35) q u i n i n e ,quinidine

24pulmonary wadge pressure

(64-67)

3. hyaline membrane (68)

jugular veincentral venous pressure (CVP) pulmonary wedge pressure free oxygen

7/21/2019 2007 Malaria

56/108

-- 55 -

radicles adult respiratorydistress syndrome

(69)

28%

creatinine 3 ./. creatinine 5 (27)

creatinine 3 ./.

BUN 40 . / . creatinine BUN 10%

hypercatabolism metabolic acidosis

(haemodialysis) (exchange transfusion) peritoneal dialysis

peritonealdialysis

h y p o v o l a e m i a , i n t r a v a s c u l a r

haemolys is , renalarteriole over activity sympathetic nervous system vasoact ive pept ide (70-75)

7/21/2019 2007 Malaria

57/108

-- 56 -

1.

2.malaria toxin toxin

3.DICDICfibrin

a c u t eglomerulonephritis

(76-78) albumin 1 24 RBC cast

Spitz.. 1946 950 acute glomerulonephritis

renalfailure(79-81)

acutetubular necrosis (ATN)

(82)

ATN ATNATN

20% (83)

(schizont) 320 28% 26%

7/21/2019 2007 Malaria

58/108

-- 57 -

320 3 units

(84)ferritin folate

ThalassemiaHaemoglobin S, E, H 2

1.

schizogony

.. 1966(85-86)

Zuckerman

51 Cr(87)

Zuckerman (autoimmune hemolysis) Facer .. 1979, 1980(88-89)

directantiglobulin test (DAT) antibodyIgM IgG

AbdallaWeatherall.. 1980 1982

(90-91)

antibodyantibody

radioimmunoassay IgG(quan t i ta t i ve )

7/21/2019 2007 Malaria

59/108

-- 58 -

complement 1 d i r e c tantiglobulin test14% IgG

IgG 1 autoimmune

hemolysis 2

1.macrophage (increase macrophage phagocytic

activity).. 1970 Sheagren reticuloendothelial system

macr ophagephagocytosis macrophage Fc receptor (92) macrophage

(93-95)

2.(increase splenic trapping) . . 1981 Wy le r

. . 1984Cranston

(95)

membrane reticuloendothelial system2.

reticulocyte

7/21/2019 2007 Malaria

60/108

-- 59 -

reticulocyte 3-4 fo la te fo la te ironparticle serumferritin

serum ferritin1

red ce l lp r e c u r s o r cytoplasm nucleus dyserythropoiesis

(96-98)

myeloid erythroid

erythroid series myeloidseries dyserythropoiesis (98)

dyserythropoiesis(99-100) (jaundice)

total bilirubin 3 ./.

(16) non-obstruct ive jaundice indirect bili rubin t o t a l b i l i r u b i n a l k a l i n ephosphatase 3 e n z ymetransaminase 2-4

10

cirrhosis (27)

7/21/2019 2007 Malaria

61/108

-- 60 -

(17 18) malaria pigment malaria pigment glycogen (19) mitochondria (101-102)

2

CPK toxin

(62)vasoconstrictionliver blood flow

liver blood flow liver bloodflow (103 -104 ) sequestration

amino

ac id , , mucosa (104)

endotoxin

7/21/2019 2007 Malaria

62/108

-- 61 -

1. 80% 3 (105)

2.

8% hypoglycemia(106)hypoclycemia50% hypoglycemia (asymptomatic

hypoglycemia)(107)

glycogen reserve glycogen glucosegluconegenesis

3. p r o t e i n serum albumin

underlying disease cirrhosis hypoalbuminemia (108)

4. ( t o x i c s u b s t a n c e endo tox in )

endotoxinlimulus test endotoxin endotoxin

67% 21.4% endotoxin endotoxin

5. (109)

mucosa(110)(111)

7/21/2019 2007 Malaria

63/108

-- 62 -

(27) recrudescence

6. mucosa g r amnegative bacteria

40% 50% (27 )

aspiration pneumonia

(32) (grand mal seizure)

(febrile convulsion) 3,000 7.7%

(112)

epilepsy

41

vasoconstriction

38.5 . 39.5-42 .

7/21/2019 2007 Malaria

64/108

-- 63 -

heatstroke neurological dificit

40 ./.

3

1.

100,000 . .lactic acidosis

2.3.

8% 144

glucose 5%

quinine

3

50% quinine 80%1-4 quinine

(asymptomatic hypoglycemia)fatty acid

7/21/2019 2007 Malaria

65/108

-- 64 -

( s t a r v a t i o nhypoglycemia)

(114)

quinine

50% 40 ./.

13.3% 66.6% (115 )

ch lo roqu ine chloroquineinsulin (116)

4 mediatormacrophage

(117) 50-90% glucose

20 (118)glycogen glucose glycogen alaninelactatehepatic gluconeogenesis metabolism

endotoxin macrophagemediator insulin quinie

quinine quinidinecell insul in C-peptide insulin

ch lo roqu ine , su l f ados ine -pyrimethamine mefloquinequinine Hughes

7/21/2019 2007 Malaria

66/108

-- 65 -

.. 1925 ..1937 quinine

lactic acidosis

4 po orprognosis (118) lact ic acidosis hydrogen ion

lactic acid 3

1. g l u cose anaerobic glycolysis lactic acid lactic acid

2.hypoxia sequestration anaerobric glycolysis lactic acid lactate CSF

3. lactate

lactate

1.

pH7.20

2.anion gap [Na+ -(CI- +HCO3-)] 10-12mEq\

7/21/2019 2007 Malaria

67/108

7/21/2019 2007 Malaria

68/108

-- 67 -

IgG (123)

2. (consumptive coagulopathy DIC)

5% f ib r inogen f ibr indegration product 20 / .

p r o t hr o m b i n p a r t i a lthromboplastin time

3. (druginduced thrombocytopenia)

4.(platelet aggregation)

DIC D IC ( d i s sem ina t ed

intravascular coagulation)

DIC

cortricosteroid, dextran heparin fibrinogen fibrinogen

turnover rate FDP(124-128)

174

7/21/2019 2007 Malaria

69/108

-- 68 -

prospective fibrinogen (actue phaseprotein response ) FDP 20 /. prothrombin t ime (PT) partial thromboplastin time (PTT) 18% 31% antithrombin 111 9 %DIC

thrombin

DIC

DIC fibrinogenturnover rate

FDP

(septicemia)

(129) aspiration pneumonia (pressure sore) CVP

(130)

"a lg id ma la r ia " (131)

7/21/2019 2007 Malaria

70/108

-- 69 -

(27,32)endotoxin

endotoxin macrophage mediator cachectin(tumour necrosis factor: TNF) plasma volume

plasma volume r e n i n aldosterone (103)

generalisedvasodilatation

haemody namicfunc t i on

cardiac index systemicvascular resistance

arrhythmia metabolic acidosis, hypoxia

(111) alpha-blocking agent(phenoxybenzamine)

2 cr eatinineclearance

(haemoglobinuria, black water fever)

(20) ( s e q ue s t r a t i o n )

haemoglobin 100/haptoglobin haemoglobin albumin methemalbumin hemoglobin

7/21/2019 2007 Malaria

71/108

-- 70 -

g lom eru larmembrane oxyhaemoglobinmethemoglobin

1.G6PD (classicalblack water fever) (133-136)o

G6PDo

o

quinineo

o

o10-40%

(109)o

chloroquine

proguanil mepacrine

quinine( d r u g i n d u c e d a u t o i m m u n e

haemolysis) stress

G6PD

2. G6PD

o x i d a t i o n primaquine (fava) druginduced hemolysis

( 1 3 8 )

2G6PD G6PD G6PD

7/21/2019 2007 Malaria

72/108

-- 71 -

primaquine G6PD classical black water fever

quinine

(sequestration)

2-3

(asexual form)(thick and thin film) Field's stainWright's stain Giemsa'sstain

(3)

2

1. (Antimalarial Agents)

(23)

2.

1. diazepam 0.2-0.4 ./.

7/21/2019 2007 Malaria

73/108

-- 72 -

3-5 5-10 phenobar-bitone 3.5-5./. (NaF) 50%

(139-141)

(semi -prone position) aspiration

pneumonia(postural pneumonia)

2.

(26) acetaminophendipyrone

agranulocytosis 42 oC

3. 80 50% 1-2 /

rebound effect 50%hypertonic solution glucagon 10

diazoxidesomatostatin

50% 5-10%

7/21/2019 2007 Malaria

74/108

-- 73 -

(NGtube)

4.

corticosteroid (alkalinization) myog lob inu r ia

( h emog l ob i n s)

5. 21%

(142)21% packed red cell

furosemide 20-40

6. DIC (disseminatedintravascular coagulation)5-92-3

(replacement therapy)10-30 heparin

(143)

7. lactic acidosis tissue anoxia

lactate

7/21/2019 2007 Malaria

75/108

-- 74 -

dichloroacetate lacticacidosis (143)

8. (supportivetreatment)

(144-146)

9. 2

adult respiratory distress syndrome

(ARDS) neutrophil, monocytemacrophage ARDSmediators

cachec in tum ournecrosis factor (TNF) ARDS (147)

positive-end expiratory pressure(PEEP) volumeloadrotatingtourniquet ap reso l i ne d iazox ide nitroprusside

10. 2 acute tubular necrosis

(148)

pre-renal

7/21/2019 2007 Malaria

76/108

-- 75 -

azotemia hemodialysis11. 10% 12.

100,000 / 40%.. 1974(exchange transfusion) .. 1979

2continuous flow cell separator10-27 1-75%

(par t ia lexchange transfusion)

4-5

7/21/2019 2007 Malaria

77/108

-- 76 -

2

1.

(chemoprophylaxis)

2.(mechanical measures)

(7)1.(anti adult measure)

(residual spraying)

2.(impregnated mosquito bednet) (33)10% 0.3

3.(antilarval measure)

7/21/2019 2007 Malaria

78/108

-- 77 -

9. Breasted JH. "The Edwin Smith Surgical

Papyrus." Chicago, Illinois: University of

Chicago Press. 1930.

10.Boyd MR, ed. Malariology. Philadelphia.

Pennsylvania: 1949.

11.Coatney GR. The simian malarias:

Zoonoses, anthroponoses, or both? Am J

Trop Med Hyg 1971; 20: 795-803.

12.Suizer AJ, Cantella R, Colichon A,

Gleason NN, Walls KW. A focus of

hyperendemic Plasmodium malariae, P.

vivax with no P. falciparum in a primitive

population in the Peruvian Amazonian

jungle: Studies by means of

immunofluorescence and blood smear.

Bull WHO 1975;52: 273-8.

13.Jarcho S. Some observations on disease

in pre-historic North America. Bull Hist

Med 1964: 38:1-18.

14.WHO fact sheet. http://www.who.int.

15. 2544. .

. ISSN 0857-6459. 1- 63.

16. . .. (). . ISBN 974-89466-2-2.2540:1-11.

17.. . ,

(). . ISBN 974-588-520-7. 2539:86-100.

10. Bunnag D, Karbwang J, Thanavibul A, et

al.High dose of primaquine in primaquine

resistant vivax malaria. Trans R Soc

Trop Med Hyg 1994;88:218-9.

11. Wilairatana P, Silachamroon U, Krudsood

S, et al. Efficacy of primaqine regimens

for primaquine-resistant Plasmodium

vivax malaria in Thailand. Am J Trop

Med Hyg 1999;61(6):973-7.

12.Buchachart K, Krudsood S, Singhasivanon

P, et al. Effect of primaquine standard

dose (15 mg/day for 14 days) in the

treatment of vivax malaria patients

inThailand. Southeast Asian J Trop Med

Public Health 2001;32(4):720-6.

13. Wilairatana P, Silachamroon U, Krudsood

S, et al. Efficacy of primaquine

regimens for primaquine-resistant

Plasmodium vivax malaria in Thailand.

Am J Trop Med Hyg 1999;61(6):973-7.

14.Silachamroon U, Krudsood S,

Treeprasertsuk S, et al. Clinical trial of

oral artesunate with or without high-dose

primaquine for the treatment of vivax

malaria in Thailand. Am J Trop Med

Hyg 2003;69(1):14-8.

15. Harrison BA, Rattanarithikul R, Peyton EL,

Mongkolpanya K. Taxonomic changes,

revised occurrence records and notes on

the Culicidae of Thailand and

neighboring countries. Mosquito

Systematics. 1990;22(3):196-227.

16. Cranston WL. Temperature regulation.

Br. Med J 1996;3:69-75.

17. Harinasuta T, Bunnag D. The clinical

features of malaria In:Malaria Principles

and Practice of Malariology.

Wernsdorfer WH & McGregor I eds.

Churchill Livingstine 1988:709-734.

18. Harinasuta T, Looareesuwan S, Bunnag

D. Cerebral malaria in Thailand :

Clinical manifestations and the

7/21/2019 2007 Malaria

79/108

-- 78 -

management. Mosquito Borne Dis Bull

1985;1:49-55.

19. White NJ, Looareesuwan S. Cerebral

malaria In: Infections of the Nervous

System, PGE Kenedy & RT Johnson ed:

Butteworths, London 1987:118-148.

20. Warrell DA, Looareesuwan S, Warrell MJ,

Kasemsarn P, Intraprasert R, Bunnag D,

Harinasuta T. Dexamathasone proves

deleterious in cerebral malaria: A

double-blind trial in 100 comatose

patients . N Engl J Med

1982;306(6):313-319.

21. Senanayake N. Delayed cerebelar

ataxia: a new complication of falciparum

malaria? Br Med J 1987;294:1253-1254.

22. White NJ, Warrell DA, Chanthavanich P,

Looareesuwan S, Werrell MJ, Hrishma

S, Wilaiamson DH, Turner RC. Severe

hypoglycemia and hyperinsulinaemia in

falciparum malaria. N Engl J Med

1983:309(29):61-66.

23. White NJ, Warrell DA, Looareesuwan S,

Chanthavanich P, Phillips RE, PongpaewP. Pathophysilogical and prognostic

significance of CSF lactate in cerebral

malaria. Lancet 1985;1:776-778.

24. amiton DK &Pikacha D. Ruptured spleen

in a malarious area. Australia & New

Zealand J Surg 1982;52:310-313.

25. Jian JB & Yu SY. An observation on

multiple infections of Plasmodium vivax

multinucleatum in its late stages. Acta

Scientiarum Naturalium Universitatis

Sunyatseni 1965;3:361-365.

26. Looareesuwan S, White NJ, Chittamas S,

Bunnag D, Harinasuta T. High rate of

Plasmodium vivax ralapse following

treatment of falciparum malaria in

Thailand. Lancet 1987;2:1052-1054.

27. White NJ, Looareesuwan S. Cerebral

malaria. In. Infections of the Nervous

System, PGE Kenedy & RT Johnson

eds. Butterworths. Lend 1987; 118-114.

28. World Health Organization. Division of

Control of Tropical Diseases: Severe

and Complicated Malaria. Trans R Soc

Trop Med Hyg 1990;84 (supplement

2):1-65.

29. World Health Organization. Severe

falciparum malaria. Trans R Soc Trop

Med Hyg 2000;94(suppl. 1):S1-S90.

30. Warrell DA, Looareesuwan S, Warrell MJ,

lntaraprasert R, Kasemsarn P, Bunnag D,

Harinasula T. Dexamethasone proves

deleterious in cerebral Malaria: a double

blind trial in 100 comatose patients. N

Engl J Med 1982; 306:313-319.

31. Teasdale G, Jennett B. Assessment of

coma and impaired consciousness: a

practical scale. Lancet 1974;2:81-84.

32. WHO Malaria Action Programme. Severeand complicated malaria. Trans R Sc)c

Trop Med Hyg 1990; 80 (suppi): 1 - 50.

33. Looareesuwan S, Warrell DA, White NJ,

Chanthavanich P, Warrell MJ,

Chantaratherakitti S, Changswek S,

Chongmankongcheep L& Kanchanaranya

C. Retinal haemorrhage, a common

physical sign of prognostic significance in

cerebral malaria. Am J Trop Med Hyg

1983, 32: 911-915.

34. Spitz S. Pathology of acute falciparum

malaria. Milit Surg 1946; 99: 555 - 572.

35. MacPherson GG, Warrell MJ, While NJ,

Looareesuwan S & Warrell DA. Human

cerebral malaria: a quantitative

7/21/2019 2007 Malaria

80/108

-- 79 -

ultrastructure analysis of parasilized

erythrocyte sequestration. Am J Path

1985; 119:385-401.

36. Udeinya 1, Schmidt JA, Aikawa M, Miller

LH & Green 1. Falciparum malaria-

infected erythrocytes bind to cultured

human endothelial cells. Science 1981 ;

213:555-557.

37. Schmid AH, Cerebral malaria: on the

nature and significance of vascular

changes. Eur Neurol 1974; 12:197-208.

38. Yoeli M. Cerebral malaria the quest for

suitable experimental models in parasitic

diseases of man. Trans R Soc Trop Med

Hyg 1976. 70:24-35.

39. Schmidt JA, Udeinya IJ, Leech JH.

Plasmodium falciparum malaria. An

amelanotic melanoma cell line bears

receptors for the knob ligand on infected

erythrocytes. J Clin Invest 1982, 70:379-

382.

40. Udeinya IJ, Miller LH, McGregor JA,

Jensen JB. Plasmodium falciparum strain

-specific antibody blocks binding ofinfected erythrocytes to amelanotic

melanoma cells. Nature 1983; 303:429-31.

41. Aley SB, Sherwood JA & Howard RJ.

Knob-positive and knob-negative

Plasmodium falciparum differ in expression

of a strain-specific malarial antigen on the

surface of infected erythrocytes. J Exp

Med Hyg 1984; 160:1585-1590.

42. Leech JH, Barnwell JW, Aikawa M, Miller

LH & Howard RJ. Plasmodium falciparum

malaria: association of knobs on the

surface of infected erythrocytes with a

histidine-rich protein and the erythrocyte

skeleton. J Cell Blot 1984; 98:1256-1264.

43. Lawler J. The structural and functional

properties of thrombospondin. Blood 1986,

67:1197-1209.

44. Raventos-Suarez C, Kaul DK, Macaluso

F, Nagel RL. Membrane knobs are

required for the microcirculatory

obstruction induced by Plasmodium

falciparum infected erythrocytes. Proc

Nati Acad Sci USA 1985; 82:3829-3833.

45. David PH, Hommel M, Miller LH, Udeinya

IJ & Oligino LD. Parasite sequestration in

Plasmodium falciparum malaria: Spleen

and antibody modulation of cytoadherence

of infected erythrocytes. Proc Nati Acad

Sci (USA) 1983; 80:5075-5079.

46. Barnwell JW, Ockenhouse CF & Knowles

DM. Monoclonal antibody OKM5 inhibits

the in vitro binding of Plasmodium

falciparum infected erythrocytes to

monocytes; endothelial and C32

melanoma cells. J lmmunol 1985;

135:3494-3497.

47. Singh B, Ho M, Warrell DA,

Looareesuwan S, Hommel M. Plasmodiumfalciparum: inhibitionlreversal of

cytoadherence of Thai Isolates with local

immune sera, Clin Exp Jummunol 1988;

72:145-1 50.

48. Cohen S, McGregor IA & Carrington S.

Gamma-globulin and acquired immunity to

human malaria. Nature 1961; 192:733-

737.

49. White NJ, Warrell DA, Looareesuwan S,

Chanthavanich P, Phillips RE, Pongpaew

P. Pathophysiological and prognostic

significance of CSF lactate in cerebral

malaria. Lancet 1985; i:776-778.

50. Warrell DA, White NJ, Veall N,

Looareesuwan S, Chanthavanich P,

7/21/2019 2007 Malaria

81/108

-- 80 -

Phillips RE, Karbwang J, Pongpaew P.

Cerebral anaerobic lycolysis and reduced

cerebral oxygen transport in human

cerebral malaria. Lancet 1988; 2:534-538.

51. Miller LH, Usami S & Chien J. Alteration

in the rheologic properties of Plasmodium

knowlesi infected red cells. A possible

mechanism for capillary obstruction. J

Clin Invest 1972; 50:1451-1455.

52. Lee MV, Ambrus JL, De Souza JM, Lee

RV, Diminished red blood cell

deformability in uncomplicated human

malaria. A preliminary report. J Cell

Physiol 1986; 114:245-251.

53. Looareesuwan S, Ho M, Wattanagoon Y,

White NJ, Warrell DA, Bunnag D.

Harinasuta T, Wyler D. Dynamic

alteration in splenic function during acute

falciparum malaria. N Engl J Med 1987;

317(1 1) 675-679.

54. Knisely MH, Stratman-Thomas WK &

Elliot TS. Observations in circulating

blood in the small vessels of internal

organs in living Macaca rhesus infectedwith malarial parasites. Anat Rec 1941;

79:90.

55. Knisely MH, Bloch EH. Microscopic

observations of intravascular

agglutination of red cells and

consequent sludging of the blood in

human diseases. Anat Rec 1942;

82:426.

56. Knisely MH, Stratman-Thomas WK &

Elliot, T.S. & Bloch EH. Khowfesi

malaria in monkeys. 1. Microscopic

pathoioical circulatory physiology of

rhesus monkeys during acute

Plasmodium knowlesi malaria in

monkeys (a motion picture). J Nati

Malaria Soc 1945: 4:285-300.

57. Onabanio AO, Maegraith BG.

Pathological lesions produced in the

brain by kallikrein (kininogenase) in

macaca mulatta infected with

Plasmodium knowlesi. Ann Trop Med

Parasitol 1970; 64:237-347.

58. Cranston WI. Temperature regulation. Br

Med J 1966; ii: 69- 75.

59. Oja -Amaize EA, Salimono LS, Williams

AI, Akinwolere OA, Shabo R, Aim GV,

Wigzeii H. Positive correlation between

degree of parasiteamia, interferon friars

and natural killer cell activity in

Plasmodium falciparum infected children.

J lmmunol 1984; 127:2296-300.

60. Klebanoff SJ, Vadas MA, Harian JM,

Sparks LH, Camble JR, Agosti JM,

Waltersdorph AM. Stimulation of

neuirophils by tumor necrosis factors. J

fmumnol 1986; 136:4220-4225.

61. Nawroth PP, Bank 1. Handley D,

Cassimeris J, Chess L, Stern D. Tumornecrosis factor/cachectin interacts with

endothelial cell receptors to induce

release of interieukin 1. J Exp Med

1986; 163: 1363-1375.

62. Wilairatana P & Looareesuwan S.

APACHE II scoring for predicting

outcome in cerebral malaria. J Trop

Med Hyg 1995;98: 256-260.

63. Hall AP, Charoendhum D, Sonkom P.

Pulmonary edema due to fluid overload

in falciparum malaria. Annual Progress

Report of the South East Asian Trebty

Organization Medical Research

Laboratories 1974175; p@ 226-233.

7/21/2019 2007 Malaria

82/108

-- 81 -

64. Duarte MIS, Corbett CEP, Boulos M,

Amato Neto V. Ultrastructure of the lung

in falciparum malaria. Am J Trop Med

Hyg 1985; 34:31-35.

65. Martell RA, Kallenbach J, Zwi S.

Pulmonary oedema in falciparum

malaria. Br Med J 1979; i:l 763-1 764.

66. James. Pulmonary damage associated

with falciparum malaria. Ann Trop Med

Hyg 1985; 79:123-138.

67. Brooks MH, Kiel FW, Sheehy TW & Berry

KG. Acute pulmonary edema in

falciparum malaria. N Engl J Med 1968;

279:732-737.

68. Rocker GM, Wiseman MS, Pearson D,

Shale DJ. Diagnostic criteria for adult

respiratory distress syndrome : time for

reappraisal. Lancet 1959; 1:120-140.

69. Till GO, Johnson KJ, Kunkel R, Ward PA,

lntravascular activation of complement

and acute lung injury. Dependency on

neuirophils and toxic oxygen

metabolises. J Clin Invest 1982;

69:1126-1135.70. Sitprija V, Indraprasit S, Pochanugool C,

Benyajati C, Piyaratn P. Renal function

in malaria. J Trop Med Hyg 1966;

69:242-246.

71. Sitprija V, Miller LH, Tisher CC,

Makaranond P, Canfield C, Johnston

EH, Pyaratn P. Renal pathology in acute

malaria : a biopsy study. J Trop Med

Hyg 1969; 72:309-31 1.

72. Sitprija V, indraprasit S, Pochanugool C,

Benyajati C, F4yaratn P. Renal failure in

malaria. Lancet 1967; 1:185-188.

73. Sitpfija V, Vongsthongsri M,

Poshyachinda V, Arthachinta S. Renal

failure in malaria: a Pathophysb"ic study.

Nephron 1977; 18:277-287.

74. Sitpriia V. Nephropathy in falciparum

malaria. Kidney Ant 1988;

75. Stone WJ, Hanchelt JE, Knepshield JH.

Acute renal insufficiency due to

falciparum malaria. Arch Intern Med

1972; 129:620-628.

76. Holienberg NK, Adams DF, Oken DE,

Abrams HL, Merriti JP. Acute renal

failure due to nephrotoxins: renal

haem~amic and arigiographic studies in

man. N Engl J Med 1970: 282:1329-

1334.

77. Holienberg NK, Epstein M, Rosen SM.

Acute oliguric renal failure in man :

evidence for preferential renal cortical

ischemia. Medicine 1%8; 47:455-74.

78. Skirrow MB, Chongsuphajaisiddhi T,

Maegriath BG. The circulation in

malaria. Ann Trop Med Parasitol 1964;

58:502-507.

79. Futrakul P, Boonpucknavig V,

Boonpucknavig S, Mitrakul C,Bhamarapravati N. Acute

glomeruionephritis complicating

Plasmodium falciparum infection. Clin

Pediatr 1974; 13:281-283.

80. Boonpucknavig V & Sitprija V. Renal

disease in acute Plasmodium falciparum

infection in man. Kidney lnt 1979;

16:44-52.

81. Parblani A, Cameron JS. Experimental

nephritis associated with Pasmodium

infection in mice. Kidney lnt 1979;

16:53-63.

82. Canfield CJ, Miller LH, Bartelloni PJ,

Eichier P, Berry KG. Acute renal failure

in Plasmodium falciparum malaria.

7/21/2019 2007 Malaria

83/108

-- 82 -

Treatment by Per loneal dialysis. Arch

Intern Med 1%9, 122:199-203.

83. Gilles HIM, Lawson JB, Sibela M, Voller

A, Alien N. Malaria anaemia and

pregnancy. Ann Trop Med Parasitol

1969; 63:245-265.

84. Looareesuwan S, Phillips RE, White NJ,

Kietinun S, Karbwang J, Rackow C,

Turner RC & Warrell DA. Quinine and

severe falciparum malaria in late

pregnancy. Lancet 1985; ii:4-8.

85. Woodruff AW, Andsell VE & Pettitt LE.

Cause of anaemia in malaria. lancet

1979.: 1:1055-1057.

86. Weatherall DJ & Abdallah S. The

anaemia of Plasmodium falciparum

malaria 8, Med Bull 1982; 38:147-151.

87. Looareesuwan S, Merry AH, Phillips RE,

Pleehachinda R, Waltanagoon Y. Ho M

Charoeniarp P, Warrell DA, Weatherail

DJ. Reduced erythrocyte survival

following clearance of malaria

parasitaemia in Thai Patients. Br J

Haematol 1987 ' 67:473-478.88. Abdallah S, Weatherall DJ,

Wickramasinghe SN, Hughes M. The

anaemia of P. falciparum malaria. Br J

Haematol 1980; 46:171,183.

89. Abdaliah 9, Weatherall DJ. The direct

antigiobulin test in P. falciparum malaria.

Br J Haematol 1982; 51:415-425.

90. Merry AH, Looareesuwan S, Phillips RE,

Chanthavanich P, Supanaranond W,

Warrell DA, Weatherall DJ. Evidence

against immune haemolysis in

falciparum malaria in Thailand. Br J

Haematol 1986; 64:187-194.

91. Sheagren JN, Tobie JE, Fox LM & Wolff

SM. Reticuloendothelial system

phagocytic function in naturally acquired

human malaria. J Lab Clin Med 1970;

75:481-487.

92. Ward KIN, Warrell MJ, Rhodes J,

Looareesuwan S & White NJ. Altered

expression of human monocyte Fc

receptor in Plasmodium falciparum

malaria. Infect lmmun 1984; 4:623-626.

93. Newsome F. Increased phagocylosis of

non-parasitized red cell in Plasmodium

bergheii malaria. Ann Trop Med

Parasitol 1984; 78: 323-325.

94. Wyler DJ, Quinn TC, Chen LT.

Relationship of alterations in spienic

clearance function and microcirculation

to host defence in acute rodent malaria.

J Clin Invest 1981; 67:1400-1404.

95. Cranston HA. Boyian CW, Carroil GL,

Sutera S, Williamson JR, Gluzman IY &

Krogstad DJ. Plasmodium falciparum

maturation abolishes physiologic red cell

deformabil ity. Science 1984; 223: 400-

402.

96. Srichaikui T, Panikbuir N & Jeumtrakul P.Bone Marrow changes in human

malaria. Ann Trop Med Parasitof 1967;

61:40-51.

97. Srichaikul T. Hematologic changes in

human malaria. J Med Assac Thai

1973; 56:658-671.

98. Wickramasinghe SN, Phillips RE,

Looareesuwan S, Warrell DA, Huges L+.

The bone marrow in human cerebral

malaria: parasite sequestration within

sinusoids. Br J Haematof 1987; 66:295-

306.

99. Wickramasingh SN, Looareesuwan S,

Nagachinta B, White NJ.

Dyserythropoiesis and ineffective

7/21/2019 2007 Malaria

84/108

-- 83 -

erythropoiesis in Plasmodium vivax

malaria. Sr J Haematol 1989; 72:91-99.

100. Looareesuwan S, Bunnag D.

Pathogenesis of anaemia in malaria ant

management. J Parasit Trop Med Ass

Thai 1988; 11(2): 35-39.

101. De Brito T, Barone AA, Faria RA.

Human Liver biopsy in P. fatciparum and

P. vivax malaria. A light and electron

microscopy study. Virchows Arch Abst

A. Pathologische Anatomie 1969;

348:220-9.

102. Corcoran TE, Hegstrom GJ, Zoeckler

SJ, Keil PG. Liver structure in non fatal

malaria. Gastroenterology 1953; 24:53-

62.

103. Fletcher A & Maegraith B. The

metabolism of the malaria parasite and

its hosts. Adv Parasitol 1972; 10:31-48.

104. Molyneux ME, Looareesuwan S,

Menzies 1, Grainger S, Phillips RE,

Wattanagoon Y, Thompson RPH,

Warrell DA. Reduce hepatic blood flow

and intestinal malabsorption in severefalciparum malaria. Am J Trop Med Hyg

1989; 40(5):470-476.

105. While NJ, Looareesuwan S, Warrell DA,

Warrell MJ, Bunnag D, Harinasuta T.

Quinine pharmacokinetics and toxicity in

cerebral and uncomplicated falciparum

malaria. Am J Med 1982; 73:564-572.

106. While NJ, Looareesuwan S, Warrell DA,

Warrell MJ, Chanthavanich P, Bunnag

D, Harinasuta T. Quinine loading dose in

cerebral malaria. Am J Trop Med Hyg

1983: 32(1):1-5.

107. Bygberg IC & Laung C. Septicemia as a

complication of falciparum malaria.

Trans R Soc Trop Med Hyg 1982;

76:705.

108. Sinton JA, Hughes TA. The functional

capacity of the liver in malaria. indian J

Med Res 1924; 12:409-422.

109. Segal HE, Hall AP, Jewell JS. Peariman

EJ, Na-Nakorn A, Mettaprakong.

Gastrointestinal function, quinine

absorption and parasite response in

falciparum malaria. Southeast Asian J

Trop Med Public Health 1974; 5:499-

503.

110. Oisson RA, Johnsion EH.

Histopathologic changes and small

bowel absorption in falciparum malaria.

Am J Trop Med Hyg 1969, 18: 355-359.

111. Karney WW, Tong MJ. Malabsorption in

Plasmodium falciparum malaria. Am J

Trop Med Hyg 1972, 121:1-5.

112. Wattanagoon Y, Srivilairit S,

Looareesuwan S, Bunnag D, Harinasuta

T, White NJ. Convulsions in falciparum

malaria. Xllth International Congress for

Tropical Medicine and Malaria, PA Kaker& AM Polderman eds. Excerpia Medica

ICS 810 ISBN, 0444810277. 1988

Abstract 258.

113. White NJ, Miller KD, Marsh K, Herry CD,

Turner RC, Williamson DH, Brown J.

Hypogiycaemia in African children with

severe malaria. Lancet 1987, i:708-712.

114. Metzger BE, Vileisis RA, RAvnikov V,

Freinkel N. "Accelerated starvation" and

the skipped breakfast in late normal

pregnancy. Lancet 1982; i 588-592.

115. Phillips RE, Looareesuwan S, White NJ,

Chanthavanich P. Karbwang J,

Supanaranond W, et al. Hypoglycemia

and antimalarial drugs: quinidine and

7/21/2019 2007 Malaria

85/108

-- 84 -

release of insulin. Br Med J 1986;

292:1319-1321.

116. Yeiich MR, Fiikins JP. Mechanism of

hyperinsulinemia in endotoxicosis Am J

Physiol 1980; 239:1 5&1 61.

117. Hughes TA. Effects of quinine on the

sugar of the blood. Indian J Med Res

1925; 13:321-336.

118. Jensen MD, Conley M, Heistowski LD.

Culture of Plasmodium falciparum: the

role of pH, glucose and lactate. J

Parasitol 1983; 69:1060-1067.

119. Hypovolaemia in PlasMedium coatneyi

malaria Ann Trop Med Parasitol 1968;

62:218-232.

120. Horstmann RD. Dietrich M, Bienzle U,

Rashe H. Malaria induced

thrombocytopenia. Bfut 1981, 42 257-

164.

121. Skudowitz RB. Katz J, Lurie A, Levin J,

Metz J. Mechanisms of

thrornbocytopenia in malignant tertian

malaria. Br Med J 1973 ii:515-518.

121. Kelton JG, Keystone J, Moors J, et al.Immune-mediated thrornbocytopenia of

malaria. J Clin Invest 1983; 71:832-836.

123. Looareesuwan S, Lee SH, Nagachinta

B, Davis J, White NJ. Throm

bocytopenia in malaria; evidence against

an antibody dependent mechanism. In.

Kager PA, Polderman AM. acts. Xllth

International Congress for Tropical

Medicine and Malaria, Excerpta Medin

ICS 810, Amsterdam, Netheriand 1988.

p 121.

124. Pukrittayakamee S, White NJ, Clemens

R, Chittamas S, Karges HE, Desakorn

S, Looareesuwan S, Bunnag D.

Activation of the coagulation cascade in

falciparum malaria, Trans R Soc Trop

Med Hyg 1989, 83.

125. Essein EM, Ebbota Mi. Platelet

secretory activities in acute malaria

(Plasmodium falciparum) infection. Acta

Haematol 1983, 7: 183-188.

126. Conned M. Pathophysiology of malaria.

Hematalogic observations in human and

animal studies. Ann Ant Med 1969;

70:134-141.

127. Devakui K, Harinasuta T, Raid HA. 125

I-labelled fibrinogen in cerebral malaria.

lancet 1966; ii:886-888.

128. Goodall HB. Three aspects of

disseminated intravascular coagulation

in subtertian malaria, Trans R Soc Trop

Med Hyg 1981; 75:884-885.

129. Bygbjerg IC, Lanng C. Septicaemia as a

complication of falciparum malaria.

Trans A Soc Trop Med Hyg 1982;

76:705.

130. Mabey DCW, Brown A, Grunwood BM.

Plasmodium falciparum malaria and

salmonella infections in Gambianchildren. J Infect Dis 1987; 155:1319-

1321.

131. Sullivan J. Pernicious fever : febris

aigida and febris comalosa. Medical

Times and Gazette 1876; 1:277-279.

132.Sitprija V. Urinary excretion patterns in

renal failure due to malaria. The effects

of phenoxybenzamine in two cases.

Aust NZ J Med 1971; 1:44-48.

133. Fairley NH & Bromfield RJ. Laboratory

studies in malaria and blackwater fever.

Part 11. Blackwater fever. Part 111. A

new blood pigment in biackwater fever

and other biochemical observations.

7/21/2019 2007 Malaria

86/108

-- 85 -

Trans R Soc Trop Med Hyg 1934;

28:141-156. 307-334.

134. Maegraith BG. Pathological processes

in malaria and blackwater fever. Oxford

Biackweii Scientific 1948.

135. Maegrafth BG. Recent advances in

tropical medicine : blackwater fever.

west Afr Med J 1952; 1:4-10.

136. Glackie WK. Blackwater fever. Clin

Proc 1944, 3:272-312.

137. Ross GR. Blackwater fever in Southern

Rhodesia in retrospect. Central African J

Med 1962; 8:294-7

138. Paflich V. Glucose-6-phosphate

dehydrogenase deficiency Part 2. Tal

Asia. Clinical Haernatol 1981, 10:814.

139. Warrell DA, Looareesuwan S, Warrell

MJ, et al. Dexamethasone proves

deleterious in cerebral malaria. A

double- blind study in 100 comatose

patients. N Engl J Med 1982;

306(6):313-9.

140. White NJ, Looareesuwan S. Cerebral

malaria. In: PGE Kennedy, RT Johnson,eds. Infections of the Nervous System.

London: Butterworths. 1987:118-44.

141. White NJ, Looareesuwan S, Phillips RE,

Chanthavanich P, Warrell DA. Single

dose phenobarbitone prevents

convulsions in cerebral malaria. Lancet

1988; 2: 64-6.

142. Looareesuwan S, Davis TME,

Pukrittayakamee S, et al. Erythrocyte

survival in severe falciparum malaria.

Acta Tropica, 1991; 48:263-70.

143. Warrell DA, Molyneux ME, Beales PF.

Severe and complicated malaria, 2nd

edition. Trans R Soc Trop Med Hyg

1990;84 (suppl):1-65.

144. Wilairatana P, Riganti M, Looareesuwan

S, Punpoowong B, Srisopark P,

Charoenlarp P. Dyspepsia in acute

falciparum malaria: a clinico-pathological

correlation. Southeast Asian J Trop

Med Public Health 1992; 23(4): 788-94.

145. Wilairatana P, Looareesuwan S,

Charoenlarp P. Liver profile changes

and complications in jaundiced patient

with falciparum malaria. Trop Med

Parasitol 1994; 45: 298-302.

146. Wilairatana P, Looareesuwan S,

Kaocharoen S, Vanijanonta S,

Charoenlarp P. Gastric emptying in

acute uncomplicated falciparum malaria.

J Trop Med Hyg 1995; 98: 22-4.

147. Wilairatana P, Looareesuwan S.

APACHE II scoring for predicting

outcome in cerebral malaria. J Trop

Med Hyg 1995;98:

148. Sitprija V, Indraprasit S, Pochanugool C,

Benyajati C, Piyaratn P. Renal functionin malaria. J Trop Med Hyg 1966;69:

242-6.

149. Looareesuwan S, Phillips RE, Karbwang

J, White NJ, Flegg PJ, Warrell DA.

Plasmodium falciparum

hyperparasitaemia: use of exchange

transfusion in seven patients and a

review of the literature. Quart J Med

(New Series 75) 1990; 277:471-81.

150. . : . : ; 2537:21-26.

7/21/2019 2007 Malaria

87/108

-- 86 -

PCR:

.

(Polymerase chain reactiontechnology)

PCR

Polymerase chain reaction

(Polymerase chain reaction; PCR)

DNA polymerase RNA specific primers

(template)

(complementary)

7/21/2019 2007 Malaria

88/108

-- 87 -

1 2

1. specificprimers (oligonucleotides)

2

2. primer

3' 1 kb 3 kb primer 21-24 nucleotides

primer primerdimmer primer

secondary structure

3. deoxyribonucleoside-5'-

triphosphate 4 (dATP,dGTP, dCTPdTTP)

4.

5. DNA polymerase (thermostableDNA polymerase) Thermus aquaticus(Taq DNApolymerase) nucleotides

35-100 nucleotides/sec nucleotides 50-60 nucleotides AmplitaqTM Taq DNA polymerase VentTM

7/21/2019 2007 Malaria

89/108

-- 88 -

DNA polymerase 3-5exonuclease activity Taq DNApolymerase

PCR Thermocycler

(Polymerase chain reaction;PCR)run gel electrophoresis

ethidium bromide

PCR

PCR 3

1. Denaturation (90-95 oC)

2. Primer annealing

(Tm-2 o

C) primers

Tm = 4(G+C)+2(A+T)

primer sequence:5CGTGACTACGATCGACA3

4G, 5C, 4A, 4T

Tm = 4(4+5)+2(4+4) = 4x9+2x8

= 36+16

= 52oC

(Tm-2oC) = 52-2 = 50 oC

3. Extension 75 oC thermostable

DNA polymerase

7/21/2019 2007 Malaria

90/108

-- 89 -

3 Polymerase chain reaction1

20-30 (1)

1.PCR

7/21/2019 2007 Malaria

91/108

-- 90 -

PCR

1 1

2 2

3 4

4 8

5 16

6 32

7 64

8 128

9 256

10 512

11 1,024

20 1,048,576

30 1,073,741,824

= 2 n

n =

PCR

whole blood EDTA EDTA blood -20 oC EDTA

blood

DNA

7/21/2019 2007 Malaria

92/108

-- 91 -

DNA

PCR

nested PCR PCR

genus primers Plasmodium

species 4 species P. falciparum P. vivax

P. malariae P. ovale primers 4

PCR assay

PCR 20 l

- 5 l of PCR buffer (MgCl2, KCl,Tris-HCl, pH 8.3)

- 125 M dNTP dTTP, dGTP, dCTP, dATP

- 50 nM of primers

- 0.4 unit of Taq polymerase

- 1 l of DNA template

steriled distilled water20 l PCR

PCR

PCR

genus

Step 1 DNA (denaturation)95oC 1

Step 2 primers DNA (annealing)

58

oC 2

Step 3 Taqpolymerase DNA

primers 72

oC 2

7/21/2019 2007 Malaria

93/108

-- 92 -

24 1 l product

PCR species

Step 1 DNA (denaturation)

95

oC 1

Step 2 primers DNA (annealing) 58 oC 2

Step 3 Taqpolymerase DNA primers 72

oC 2

30 15 l product 3 % agarose gelelectrophoresis, 70 volts gel ethidium bromide destain gel gel DNA fragments

7/21/2019 2007 Malaria

94/108

-- 93 -

2 nestedPCR 2% agarose gel lane S =standard markers, lane 1= P. falciparum, lane 2= P. vivax, lane 3=

P. malariae lane 4= P. ovalae

PCR

PCR (contamination)

PCR (PCRgrade)

7/21/2019 2007 Malaria

95/108

-- 94 -

References

1. Gibbs, RA. DNA amplification by the polymerase chain reaction. Analytical

Chemistry 1990; 62:1202-1214.

2. Markham, AF. The polymerase chain reaction: A tool for molecular medicine.

British Medical Journal 1993; 306: 441-446.

3. Snounou G, Viriyakosol S, Jarra W, Thaithong S, Brown KN. Identification of

the four human malaria parasite species in field samples by the polymerase

chain reaction and detection of a high prevalence of mixed infections.

Molecular and Biochemical Parasitology1993;58:283-292.

4. Snounou G, Viriyakosol S, Zhu XP, Jarra W, Pinheiro L, Rosario VE,

Thaithong S, Brown KN. High sensitivity of detection of human malaria

parasites by the use of nested polymerase chain reaction. Molecular and

Biochemical Parasitology 1993;61:315-320.

-------------------------------------------------------------------------

7/21/2019 2007 Malaria

96/108

-- 95 -

Field (Fields stain)for Thick film

Fields stain

(thickfilm) 2Stain A (methylene blue) Stain B (eosin)

Stain A

Methylene bkue (medicinal) 0.8 g.

Azure I 0.5 g.

Disodium hydrogen phosphate

(anhydrous) 5.0 g.

Potassium hidrogen phosphate 6.25 g.

Distilled water 500 ml.

Stain B

Eosin 1.0 g.

Na2HPO4 5.0 g.

KH2PO4 6.25 g.

Distilled water 500 ml.

(dye) 24

oxidation scum

()

1.

absolute methyl alcohol fixed2. Stain A 5

3. 5 4. Stain B 3

5. 5 6.

Stain B (greenish colour)

()

7/21/2019 2007 Malaria

97/108

-- 96 -

Wright (Wrights stain) for thin film

Stock solution

Stain (powder) 0.3 g.

Glycerin (C.P.) 3.0 ml.

Methyl alcohol, absolute,

acetone free 97.0 ml.

powdered stain glycerin methylalcohol 2-3

()

1. stock solution fix 1

2. buffer (pH 7.2) 3

3.

4.

Wright (Modified Wrights stain)for thick film

Stock solution

Stain (powder) 0.3 g.

Glycerin (C.P.) 3.0 ml.

Methyl alcohol, absolute, acetone free

97.0 ml.

(

)1. stock solution 1 ml.

(pH

7.2) 2 ml.10 ml.

2. 2-5

3.

4.

7/21/2019 2007 Malaria

98/108

-- 97 -

Giemsa (Giemsas stain) for thick & thin film

Giemsa

Giemsas solution

Giemsas powder 3.8 g.

Methyl alcohol 205 ml.

Glycerine 250 ml.

Giemsa glycerine methyl alcohol (Pyrex bottle) methylalcohol alcohol incubate 37

oC 24

1. fix

2. 3 %Giemsa 3 ml. buffer 97 ml.3. 30

45

o

1.

absolute methyl alcohol 30 fix 2.

3 %Giemsa 3 ml.

buffer 97 ml.3.

staining jar 4. 30

45 o

7/21/2019 2007 Malaria

99/108

-- 98 -

(Buffered water)

Giemsa buffer pH = 7.2 -(neutral)

Stock solution

M/15 Na2HPO4 Sodium hydrogen phosphate 9.5 g. per litre

M/15 KH2PO4 Potassium dihydrogen phosphate 9.02 g. per litre

pH 7.0 pH 7.2

M/15 Na2HPO4

(ml.)

M/15 KH2PO4

(ml.)

Distilled water

(ml.)

pH 7.0 61.1 38.9 900

pH 7.2 72.0 28.0 900

pH 7.2

KH2PO40.7 g.

Na2HPO4.1.0 g.

Distilled water.1000 ml.

pH

7/21/2019 2007 Malaria

100/108

-- 99 -

7/21/2019 2007 Malaria

101/108

7/21/2019 2007 Malaria

102/108

-- 101 -

B07 Plasmodium malariae growing trophozoite (band form)B08 Plasmodium malariae growing trophozoite (compact form)B09 Plasmodium malariae growing schizontB10 Plasmodium malariae mature schizont

(rosette formation)

B11 Plasmodium malariae microgametocyte chromatin cytoplasm

B12 Plasmodium ovale growing trophozoite infected RBC

B13 Plasmodium ovale mature schizontB14 Plasmodium ovale macrogametocyteB15 Plasmodium ovale microgametocyte

Flagellates & Ciliate

C01 Trypanosoma cruzi trypomastigote Giemsa

C02 Trypanosoma gambiense trypomastigote C03 Trypanosoma cruzi epimastigote

(triatomid bug)C04 Leishmania donovani promastigote

C05 Leishmania donovani amastigote smear

GiemsaD01 Giardia intestinalis trophozoite Giemsa

D02 Giardia intestinalis cyst trichromeD03 Giardia intestinalis cystnormal saline solutionD04 Trichomonas vaginalis trophozoite GiemsaD05 Dientamoeba fragilis trophozoite trichromeD06 Balantidium coli trophozoite iron-hematoxylinD07 Balantidium coli cyst iron-hematoxylin

7/21/2019 2007 Malaria

103/108

-- 102 -

D08 Trichomonas vaginalis trophozoite

Coccidia

E01 Cryptosporidium parvum mature oocyst DMSOE02 Cyclospora cayetanensis mature oocyst DMSOE03 Isospora belli immature oocyst () mature

oocyst ()normal saline solutionE04 Toxoplasma gondii trophozoites peritoneal fluid

GiemsaE05 Toxoplasma gondii pseudocyst

Giemsa

E06 Microsporidia oocyst Gram ChromotopeE07 Sarcocystis sp. oocystnormal saline solutionE08 Sarcocystis sp. cyst

7/21/2019 2007 Malaria

104/108

-- 103 -

No. -

1. 119 9 52210

054-286-117

110086-1182-958

2. - () 83000

076-254-421-9

3 227 1 67130

056-791-211

109

4. ..

086-3052536

5. 309 3

081-021-2117

053-603114

6. . 12120

02-986-9213

7.

10600

02-475-2901

8.

02-923-9710

9. 10560

02-338-1133

114

10. 2 15/38-43 10160

02-8074044

3112

11. 3

200000

038-271-881-2

0895481597

12. 52240

087-572-6713

13. 7 6 1150

038-684444

109

No. -

7/21/2019 2007 Malaria

105/108

-- 104 -

14. 32170

044-569080

109

15. 70000

081-4556497

16

. 90110074-451562

17. . 90110

074-451562

18. 48120

042-581255-6

19. 92160

081-6315031

20. 114 1

24120 081-6381-492

21. 543 10600

02-4370123

1229

22. 81120

075-643255

117

23. 196 73000

034-254150-9

1032

24. 195 2 84260

086-6843673

077-387189

25. 6 .

40000043-222818-9

26. 85000

077-812630

147

27. 12120

02-986-9213-9

7410

28. 123 8 . 10270

02-3894915

29. 94000

073-489070

086-9579900

30. 70140

032-281858

081-8579535

No. -

31. 679 .11 10230

085-1305886

7/21/2019 2007 Malaria

106/108

-- 105 -

32. 679 .11 10230

02-5174270

1315

33. (Lab OPD) 91000

081-9575907

074-723500

34. .11 20180

038-245051

35. 21130

038-603838

116

36. 42000 081-7698316

37. 206 1 11000

086-985-7679

38. 2

081-3243430

39. () 37240

081-8405596

40. 29 70120

032-355300

496

41. 29 70120

032-355300

496

42. 35/3 6 78 10130 089-1180671

43. 32000

086-6830907

044-511757

44. () 421 93000 086-1980703

45. 42 1 84210

077-369053

086-2747334

46. 34000

086-2579750

045-244973

7/21/2019 2007 Malaria

107/108

-- 106 -

No. -

47.

25220081-7906300

48. 60 3 84250

089-7296959

077-399084

49. 10700

02-4197170

50. 86 1 94230

089-2985955

073-494037

51. 40320

043-266142-3

52. 61000

056-511081

53. 8 10120

02-2897769

54. 23140

039-591040

109

55. 86130 081-8917-934

56.

1040002-2454601-5

5008

57. 152 85000

073-212228

58. 86000

086-593-1648

59. 65110

086-935-5468

60.

34130

045-481797

106

61. 135 34170

045-491-182

1024

62. 285 . 2

053-455074

7/21/2019 2007 Malaria

108/108

No. -

63. (Lab)

93130089-732-2439

074-691031

64. 80360

075-371025

106

65. 84280

081*0841297

077-377034

66. () 53000

086-917-5094

67. - 57340053-606221-2

107

68. 44000

087-2167495

69. 37180

084-9629187

045-463114

70. 02-5903543

71.

54170083-760-1576

054-647458-9

72. 106 95110

087-290-7032

02-354-9100