Nuclear factor kappa B in urine sediment
- 1. Nuclear factor kappa B in urine sediment: a useful indicator to detect acute kidney injury in Plasmodium falciparum malaria Author: Chuchard Punsawad and Parnpen Viriyavejakul Malaria Journal 2014, 13:84 doi:10.1186/1475-2875-13-84 speaker：邱以涵 advisor：張永宗 Date: 2015.04.07 1
- 2. CONTENTS ? ABSTRACT ? MALARIA LABORTARY DIAGNOSIS ? INTRODUCE ACUTE KIDNEY INJURY ? BIOMARKERS OF RENAL FUNCTION ? METHODS ? RESULTS ? CONCLUSIONS ? TAKE HOME MESSAGE 2
- 3. ABSTRACT ? Acute kidney injury (AKI) is one of the major complications of Plasmodium falciparum malaria. ? Activation of transcription factor nuclear factor kappa B (NF-κB) involved in the development of progressive renal inflammatory diseases. 3
- 4. THE AIM of THE STUDY was to determine whether urinary sediment NF-κB p65 can act as a biomarker for AKI in patients with P. falciparum malaria. NF-κB P65 P. falciparum malaria biomarker acute kidney injury 4
- 5. MALARIA LABORTARY DIAGNOSIS ? Microscopy (morphologic analysis) This technique remains the gold standard for laboratory confirmation of malaria. ? Molecular Diagnosis PCR is for confirming the species of malaria after the diagnosis either smear microscopy or RDT. 5
- 6. ? Antigen Detection Rapid Diagnostic Test(RDTs) ? Serology Using either indirect immunofluorescence (IFA) or enzyme-linked immunosorbent assay (ELISA).
- 7. Rapid Diagnostic Test(RDTs) 1) accuracy: >99% 2) read Time: 5-10 minutes 3) easy to use and carry 4) competitive price 7
- 8. Disadvantages ? The RDT may not be able to detect some infections with lower numbers of malaria parasites.
- 10. CAUSES OF ACUTE KIDNEY INJURY hypoperfusion 30~60% 20~40% 1~10% 10
- 11. ?Renal tubular epithelial cells and muddy brown casts(granular casts) suspect acute tubular necrosis. Possible Urine Sediment Findings in Acute Kidney Injury 11
- 12. Possible Urine Sediment Findings in Acute Kidney Injury ? Urinary eosinophils suggest acute interstitial nephritis. ? RBC casts indicate glomerulonephritis or vasculitis.
- 13. Biomarkers of Renal Function Biomarker Type Biomarker Functional marker Serum creatinine Up-regulated proteins Neutrophil gelatinase-associated lipocalin (NGAL) Kidney injury molecule 1 (KIM-1) Interleukin 18 (IL-18) 13
- 14. Creatinine ? At present, serum creatinine, which is used to measure the glomerular filtration rate (GFR), is the most commonly used marker of renal function. 14
- 15. Why we search for new biomarker? ? Serum creatinine is a delayed and unreliable indicator of acute kidney injury (AKI). a) The creatinine level is influenced by multiple non-renal factors. b) Serum creatinine measurement does not allow differentiation between hemodynamically mediated changes in renal function. 15
- 16. THE AIM of THE STUDY was to determine whether urinary sediment NF-κB p65 can act as a biomarker for AKI in patients with P. falciparum malaria. NF-κB P65 P. falciparum malaria biomarker acute kidney injury 16
- 17. NF-κB P65 17
- 18. METHODS MEASUREMENT OF URINARY SEDIMENT NF- p65 LEVELS Phospho NF-κB P65 mouse monoclonal antibody Phospho NF-κB P65 rabbit monoclonal antibody OD:450nm 18
- 20. WITH AKI 20
- 21. RESULTS LEVELS OF NF-KB P65 IN THE URINARY SEDIMENT CELLS WITH AKI 21
- 22. DAY 0 DAY 7 CREATININE BLOOD UREA NITROGEN URINE WBC Positive correlation 22
- 23. ? A significant positive correlation was found between NF-κB p65 in the urinary sediment cells and serum Cr. DAY 0 DAY 7 Rs:0.9000; p:0.037 Rs: 0.975; p:<0.005 23
- 24. CONCLUSIONS ? the urinary level of NF-κB p65 has a potential role as a disease biomarker for renal tubular epithelial cells and AKI among complicated P. falciparum malaria patients. ? Early diagnosis and prompt management including dialysis can reduce mortality . 24
- 25. Research associated with Plasmodium falciparum and NF-κB p65 25
- 26. Liver changes in severe Plasmodium falciparum malaria: histopathology, apoptosis and nuclear factor kappa B expression Parnpen Viriyavejakul12*, Vasant Khachonsaksumet1 and Chuchard Punsawad3 Malaria Journal 2014, 13:106 doi:10.1186/1475-2875-13-106 ? Results Hyperplastic Kupffer cells and portal tract inflammation are two main features found in the liver tissues of severe P. falciparum malaria cases. In addition, NF-κB is associated with Kupffer cells and lymphocyte apoptosis in severe P. falciparum malaria. 26
- 27. Take home message the urinary level of NF-κB p65 has a potential role as a disease biomarker in AKI among complicated P. falciparum malaria patients. NF-κB P65 P. falciparum malaria biomarker acute kidney injury 27
- 28. 28
- 30. Recent studies show that NF-κB p65 activation has also been detected in peripheral blood mononuclear cells (PBMCs) from complicated P. falciparum malaria patients. 30
Editor's Notes
- #6: http://www.cdc.gov/malaria/diagnosis_treatment/rdt.html
- #8: malaria rapid test kit? 1)high accracy and faster results? 2)easy to use and carry? 3)competitive price? 4)ISO approved? 1)Features: high sensitivity, simple & easy to use and the result is accurate . 2)model?number:?cassette(3.0mm 4.0mm) 3)Read Time:?5-10minutes 4)Accuracy: >99%
- #11: Explanation: http://www.merckmanuals.com/professional/genitourinary_disorders/acute_kidney_injury/acute_kidney_injury_aki.html 通常它發生是因為腎組織損壞造成的，其起因為任何病因(例如低血壓)腎血流量的減少(腎缺血(ischemia))、暴露於對腎有害的物質(Nephrotoxicity)、腎發炎的過程，或阻塞尿流量的尿路阻礙。
- #12: The renal epithelial cells that makeup a renal epithelial cell cast can degrade while they are present free in the tubular lumen. In this case, they can form a pigmented or granulated cast. These are less specific than a true renal epithelial cell cast, since other structures such as red cells or white cells can also degrade and form granular casts. Usually, however, large numbers of pigmented granular casts, so called muddy brown casts, are indicative of ongoing acute tubular necrosis (ATN).
- #13: "typical" dysmorphic red cells in glomerular haematuria.
- #14: Up-regulated 正向調控
- #16: accumulation of nitrogenous products in the blood (azotemia)氨血症 Hemodynamically血液動力 1.The creatinine level is influenced by multiple non-renal factors, such as age, gender, muscle mass, muscle metabolism and diet. 2.An increase in the serum creatinine level can take several hours or days to reach a new steady state, represents a delayed indication of a functional change in GFR that occur in the kidney during the early stage of AKI. 3. Serum creatinine measurement does not allow differentiation between hemodynamically mediated changes in renal function, such as pre-renal azotemia from intrinsic renal failure or obstructive uropathy.
- #18: The source of NF-κB p65 in urine sediment possibly originated from renal tubular cells, which were detached from the basement membrane into the lumen of renal tubules and/or from the WBC. NF-?B是由p50和p65所形成的heterodimer，存在於細胞質中，平時I?B會附著在上面，抑制其活性 a. 可以啟動NF-?B活性的訊號因子有 i. 發炎激素(proinflammatory cytokine) ii. 細菌與病毒感染(bacterial and viral infection) iii. 氧化自由基(reactive oxygen species) iv. 生長刺激因子(mitogens) b. 反應路徑： i. 外界活化因子(activators)會使IKK複合體活化 ii. 活化的IKK複合體接著磷酸化I?B，使其失去與NF-?B的結合力，脫落的I?B，最後會被Proteasome/ubiquitin系統分解掉 iii. 活化的NF-?B進入細胞核與基因結合，吸引共活化因子(coactivator)促進轉錄，進而影響基因表現 NF-κB作用機制。在此圖中，將以Rel與p50蛋白組成的NF-κB異質二聚體為例。當處於激活狀態時，NF-κB位於細胞質中且與抑制蛋白IκBα形成複合體。通過內在膜受體的介導，一些胞外信號物質可激活一種稱為IκB激酶（IKK）的酶。IKK轉而磷酸化IκBα蛋白，這將導致後者的泛素化，使得IκBα從NF-κB上脫離下來，最終IκBα被蛋白酶體所降解。被激活的NF-κB接下來轉移到細胞核內，在這裡會結合到DNA上被稱為反應元件（RE）的特異性序列上。DNA/NF-κB 複合體接下來會招募其它蛋白，如輔激活物與RNA聚合酶，這些蛋白將下游的DNA轉錄為mRNA並轉而被翻譯為蛋白質，這些蛋白最終導致細胞功能發生改變
- #19: http://www.activemotif.com/catalog/355/face-nf%CE%BAb-p65-profiler
- #23: CORRELATIONS BETWEEN URINARY SEDIMENT NF-KB P65 LEVEL AND CLINICAL DATA
- #31: Data suggest that NF-κB p65 levels may reflect the degree of renal tubular damage caused by PRBC sequestration, leading to restricted local blood flow and host cytokine response, contributing to progressive renal tubular injury. ?