Biomarkers Utility for Sepsis Patients Management

Sepsis is a global problem in either developing or developed countries and it is expected that the number of patients with sepsis and septic shock will tremendously increase in next decades also because of the antibiotic resistance growing issue worldwide. Criteria for sepsis diagnosis and prognosis have been recently established, but still a further understanding of the role of biomarkers in this setting is needed. Better utilization of biomarkers such as white blood cell count, CRP, lactate, procalcitonin, presepsin and bioadrenomedullin in sepsis patients, a state of the art on how to use them is needed. This review will focus on the actual recognized role of sepsis biomarkers not only for diagnosis purpose but also to improve patients treatment results in order to reduce mortality, hospital length of stay and cost related

Differences Between Alpha-Fetoprotein (AFP) and Prothrombine Induced by Vitamin K Absence or Antagonist II (PIVKA-II) Values as Early Detection Method for Hepatocellular Carcinoma (HCC) and Cirrhosis

Background: Hepatocellular carcinoma is a common cancer worldwide and has a high mortality. Biomarkers could theoretically help to detect the disease at an earlier stage before symptoms occur and improve the treatment outcomes. The first biomarker found was AFP (not very accurate and 30-40% of HCC may be missed). PIVKA-II can be used as an early detection method to diagnose HCC.Method: A cross sectional study on in-patients or out-patients at Saiful Anwar Malang Hospital from July 2016 to October 2016.Results: The p value (p > 0.05) obtained using Kolmogorov-Smirnov was 0.166 for diagnosis of HCC and 0.147 for the diagnosis of hepatic cirrhosis. The p value (p > 0.05) obtained using Shapiro-Wilk was 0.103 for diagnosis of HCC and 0.087 for the diagnosis of cirrhosis. Comparative test using the LSD method showed PIVKA-II serum levels in HCC as compared to hepatic cirrhosis as significant with a p-value less than 0.05 (p < 0.05), that is 0.025. However comparative test using the Tukey HSD method showed that the results obtained were not significant. According to the PIVKA-II cut off value, the sensitivity and specificity to detect cirrhosis and HCC was as large as 100%. According to the AFP cut off value, the sensitivity to detect cirrhosis and HCC was 93.3% and the specificity was 76.92%.Conclusion: Both PIVKA-II and AFP can be used to detect cirrhosis and HCC. However PIVKA-II exhibited better sensitivity and specificity in the detection of cirrhosis and HCC

Exploring Systemic Lupus Erythematosus Pathogenesis through Animal Models: A Systematic Review of Humanized and Pristane-Induced Lupus Mice

Studies involving experimental animals to explore the pathogenesis of systemic lupus erythematosus (SLE) which leads to the selection of optimal therapy have been widely conducted. The well-known model used to study SLE includes the pristane-induced mouse model and the more recently developed humanized mouse model that implants human immune cells into immunodeficient mice. The current state of the research has yet to provide a systematic review that analyzes both model and its contribution to our understanding of SLE pathogenesis. This systematic review-based study aims to provide a comprehensive overview of the development and application of pristane-induced and humanized mouse models. We obtained several relevant article sources include: (1) Search Strategy, on databases such as PubMed, MEDLINE, ScienceDirect, and Cochrane by adjusting the protocols listed in the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA); (2) Eligibility based on exclusion and inclusion criteria; and (3) Data Extraction. The findings show that 30 articles are relevant to the subject matter. Several strains of mice were used in the model of the 0.5 pristane injection method and the humanized mice model. All studies showed similar patterns in the onset and manifestation of SLE in mice models with slight variations. The purpose of using the pristane injection method and humanized mice model is adjusted to the output of each study. A variety of research preferences can be used as a reason for choosing pristane and humanized cells transplanted SLE methods in making lupus model mice

Differences Between Alpha-Fetoprotein (AFP) and Prothrombine Induced by Vitamin K Absence or Antagonist II (PIVKA-II) Values as Early Detection Method for Hepatocellular Carcinoma (HCC) and Cirrhosis

Background: Hepatocellular carcinoma is a common cancer worldwide and has a high mortality. Biomarkers could theoretically help to detect the disease at an earlier stage before symptoms occur and improve the treatment outcomes. The first biomarker found was AFP (not very accurate and 30-40% of HCC may be missed). PIVKA-II can be used as an early detection method to diagnose HCC.Method: A cross sectional study on in-patients or out-patients at Saiful Anwar Malang Hospital from July 2016 to October 2016.Results: The p value (p > 0.05) obtained using Kolmogorov-Smirnov was 0.166 for diagnosis of HCC and 0.147 for the diagnosis of hepatic cirrhosis. The p value (p > 0.05) obtained using Shapiro-Wilk was 0.103 for diagnosis of HCC and 0.087 for the diagnosis of cirrhosis. Comparative test using the LSD method showed PIVKA-II serum levels in HCC as compared to hepatic cirrhosis as significant with a p-value less than 0.05 (p < 0.05), that is 0.025. However comparative test using the Tukey HSD method showed that the results obtained were not significant. According to the PIVKA-II cut off value, the sensitivity and specificity to detect cirrhosis and HCC was as large as 100%. According to the AFP cut off value, the sensitivity to detect cirrhosis and HCC was 93.3% and the specificity was 76.92%.Conclusion: Both PIVKA-II and AFP can be used to detect cirrhosis and HCC. However PIVKA-II exhibited better sensitivity and specificity in the detection of cirrhosis and HCC

Proteinuria Severity in Lupus Nephritis is Associated with Anti-dsDNA Level and Immune Complex Deposit Location in Kidney

Lupus nephritis (LN) is one of the manifestations of Systemic Lupus Erythematosus (SLE), with proteinuria being one of the clinical manifestations. The proteinuria pathogenesis is associated with anti-dsDNA antibody and the location of immune complex deposits within the kidney. This study aims to investigate the correlation of the severity of proteinuria with the location of immune complex deposits and the level of anti-dsDNA antibody in LN. Data were collected in cross-section. Fifty-three patients with LN in Saiful Anwar Hospital Malang, who underwent renal biopsy, were included. Hematoxylin-eosin staining and immunofluorescence analysis were used to assign subjects to different histopathological classes and determine the immune complex deposits. The spot urine samples were evaluated using the dipstick method for semi-quantitative proteinuria. The anti-dsDNA antibody levels were evaluated using the enzyme-linked immunosorbent assay (ELISA). Turbidity and enzymatic tests were conducted to elucidate urine protein and creatinine content, respectively. The level of proteinuria is significantly different among the different locations of immune complex based on the dipstick and protein/creatinine methods (p = 0.021 and p = 0.005, respectively). There was a significant correlation between anti-dsDNA antibody level and the severity of proteinuria (r = 0.326 based on dipstick and r = 0.28 based on protein/creatinine method). Thus, proteinuria in LN is determined by anti-dsDNA level and the location of immune complex deposits in the kidney

Development of Candidate Antigens for Rapid Test Kit to Detect Autoantibodies in Patients with Systemic Lupus Erythematosus

Systemic Lupus Erythematosus (LES) is an autoimmune inflammatory disease characterized by the formation of anti-nuclear antibodies (ANA) and anti-double stranded DNA (anti-dsDNA) antibodies as diagnostic markers. Detection of such autoantibodies requires advanced equipment and trained personnel. This study was conducted to acquire candidate antigens that can be used for rapid test kit for practical and accurate detection of ANA and anti-dsDNA to speed up SLE diagnosis. Nuclear proteins and DNA derived from cell lines, hair follicles, and leukocytes of SLE patients and healthy individuals were isolated using QiaGEN kit and modified-manual procedure. Antigen-antibody bonds were tested by dot blot assay. The strongest binding between DNA antigens of a healthy individual and antibodies occurred at dilution factors of 1:5,120 for the antigen and 1:2,560 for the antibody. The strongest binding between nuclear protein antigens from the cell line and antibodies occurred at dilution factors of 1:512 for the antigen and 1:1,600 for the antibody. Nuclear antigens derived from cell line and DNA antigens of healthy individuals were antigen candidates for the development of ANA and anti-dsDNA rapid detection tests

Pengembangan Prototipe Antigen dsDNA Untuk Kit Diagnosis Dini Lupus Eritematosus Sistemik

Lupus Eritematosus Sistemik (LES) merupakan penyakit keradangan jaringan ikat multisistem yang ditandai dengan terbentuknya autoantibodi terhadap komponen sel sehingga menimbulkan kerusakan pada jaringan. Manifestasi klinisnya sangat bervariasi, sehingga sulit untuk mendiagnosis lupus, karenanya diperlukan tes yang praktis dan akurat untuk meningkatkan kecepatan diagnosis. Salah satu aplikasi alat diagnostik yang mudah digunakan adalah metode rapid test kit, atau immunochromatographic test(ICT). Tujuan penelitian ini adalah untuk mengetahui performance kandidat antigen DNA untuk deteksi antibodi anti-dsDNA. Sampel penelitian berupa dua kelompok antigen yang berasal dari kultur cell line HeLa dan nukleus limfosit whole blood, antibodi dari serum kontrol dan pasien LES sesuai kriteria SLICC 2012. Metode yang digunakan untuk mengisolasi antigen DNA adalah dengan kit ekstraksi DNA, kemudian dilakukan Dot enzyme immunoassay (DEI) dengan uji titrasi papan catur (checkerboard), untuk menetukan konsentrasi terbaik dari antigen dan antibodi primer yang ditandai dengan densitas perubahan warna yang signifikan. Kemudian dilanjutkan DEI terhadap 24 sampel anti-dsDNA positif dan anti-dsDNA negatif, berdasarkan hasil titrasi checkerboard untuk menentukan spesifisitas dan sensitifitas intensitas warna dot blot assay terhadap anti-dsDNA hasil pengukuran dengan ELISA. Didapatkan perubahan warna yang paling menonjol pada uji checkerboard adalah pada pengenceran antigen dari limfosit whole blood 1/640(konsentrasi antigen 3,16ng/μL) dan pengenceran antibodi 1/3200. Tidak didapatkan perubahan warna yang signifikan pada uji checkerboard dengan antigen dari sel HeLa. Hasil perhitungan uji diagnostik dengan DEI pada cut off 105,23 memiliki sensitivitas 70,83%, spesifisitas 75%, nilai ramal positif(NRP) 73,91% dan nilai ramal negative(NRN) 72,00%. Diperlukan penelitian lebih lanjut tentang pengembangan antigen sel limfosit untuk pembuatan kit diagnostik anti-dsDNA

Urine Specific Proteins and Alpha-1 Antitrypsin Concentrations to Assess the Severity of Lupus Nephritis

Background. Current biomarkers for evaluating disease activity or severity in lupus nephritis (LN) are considered to be unsatisfactory. Pathological changes in glomerular basement membrane and selectivity of electrical discharge are causing specific patterns of urine proteins excretion. Together with alpha-1 antitrypsin (AAT), they are expected to become new biomarkers to assess LN activity. Method. Seventy-one urine samples were collected from healthy controls and LN patients. Patterns of urine specific proteins were determined using column chromatography and SDS-PAGE tests, LN activity was calculated using SLEDAI-renal domain score, and AAT concentrations was measured by ELISA. Result. The majority of proteins in the control group have molecular weights of >66 kDa (88%) and 21- to 25-kDa proteins were observed only in the case group. The p values for differences in urine AAT concentration between active LN and healthy controls, inactive LN and healthy controls, and active LN and inactive LN were 0.004, 0.046, and 0.054, respectively, whereas those for urine AAT/creatinine ratio were 0.489, 0.019, and 0.915, respectively. Conclusion: There were differences in the patterns of the molecular weight of proteins and urine AAT concentrations between case group and control group. However, no such differences were identified between active and inactive LN

Hubungan Rasio IL-6 Urin/Kreatinin Urin dan Eritrosit Dismorfik pada Flow Cytometry Urine Analyzer dengan Kelainan Glomerular pada Pasien Hematuri.

Latar Belakang: Kelainan glomerular merupakan penyebab tersering penyakit ginjal tahap akhir. Biopsi ginjal adalah standar baku emas menentukan kelainan glomerular, tetapi invasif dan memerlukan persyaratan khusus. Eritrosit pada kelainan glomerular berbentuk dismorfik. Urinalisis dengan flow cytometry urine analyzer diharapkan dapat menggantikan pemeriksaan mikroskopis dalam membedakan asal hematuri. Rasio IL-6 urin/kreatinin urin diketahui berkorelasi positif dengan skor jejas glomerulus. Penelitian ini bertujuan untuk mengetahui hubungan rasio IL-6 urin/kreatinin urin dan eritrosit dismorfik pada flow cytometry urine analyzer guna mengetahui apakah dapat digunakan untuk membedakan kelainan glomerular dan non glomerular. Metode: Subyek penelitian dibagi menjadi kelompok hematuri glomerular, non glomerular dan kontrol sehat. Penentuan hematuri glomerular menggunakan kriteria klinis. Penghitungan eritrosit dismorfik menggunakan flow cytometry urine analyzer dan mikroskop fase kontras. Kadar IL-6 urin diukur dengan metode ELISA dan kreatinin urin diukur dengan metode enzimatik. Hasil: Kadar IL-6 urin antara kelompok hematuri glomerular dengan non glomerular dan kelompok hematuri non glomerular dengan kontrol berbeda bermakna (p=0,014, p=0,007). Kadar IL-6 urin antara kelompok hematuri glomerular dan kontrol tidak berbeda bermakna. Rasio IL-6 urin/kreatinin urin tidak berbeda bermakna antar ketiga kelompok. Kadar IL-6 urin dan rasio IL-6 urin/kreatinin urin hematuri glomerular berkorelasi negatif dengan persentase eritrosit dismorfik pada flow cytometry urine analyzer dan mikroskop fase kontras. Persentase eritrosit dismorfik pada flow cytometry urine analyzer (cut off 51,24%) memiliki sensitivitas 96,70% dan spesifisitas 90,00%, sedangkan rasio IL-6 urin/kreatinin urin (cut off 0,135 ng/mg) memiliki sensitivitas 80,00% dan spesifisitas 23,30%, dalam membedakan hematuri glomerular dan non glomerular Kesimpulan: Persentase eritrosit dismorfik pada flow cytometry urine analyzer memiliki nilai diagnostik lebih baik dibanding rasio IL-6 urin/kreatinin urin dalam membedakan hematuri glomerular dan non glomerular

Profil Protein Spesifik Urine Dihubungkan Dengan Kelas Histopatologi Dan Aktivitas Penyakit Nefritis Lupus

viii ABSTRAK Farida, Dany, 2018, Profil Protein Spesifik Urine Dihubungkan dengan Kelas Histopatologi dan Aktivitas Penyakit Nefritis Lupus. Program Pendidikan Dokter Spesialis I Patologi Klinik Fakultas Kedokteran Universitas Brawijaya Malang. Pembimbing (1) Dr. dr. Hani Susianti, Sp.PK(K), (2) dr. I. Putu Adi Santosa, Sp.PK Nefritis Lupus (NL) merupakan manifestasi serius pada Lupus Eritematosus Sistemik (LES). Biomarker yang ada saat ini untuk mengevaluasi aktivitas NL masih belum memuaskan, biopsi ginjal masih menjadi standar baku untuk penilaian kelas histopatologi. Kelas histopatologi dan aktivitas NL digunakan untuk menentukan prognosis atau progresifitas NLl. Perubahan patologis pada membran basalis dan selektifitas protein akan menyebabkan suatu pola protein spesifik dari urin yang diekskresi. Identifikasi pola protein spesifik tersebut diharapkan membantu mengevaluasi progresifitas dan menentukan prognosis NL Tujuan penelitian ini adalah mengetahui gambaran protein spesifik urin pada NL berdasarkan kelas histopatologi dan aktivitas. Metode cross-sectional berdasarkan studi observasional dengan sampel 20 pasien NL dilakukan pemeriksaan protein urin dengan kromatografi kolom dan SDS-PAGE. Pasien NL dilakukan biopsi ginjal untuk menentukan kelas histopatologi dan aktivitasnya. Dilakukan deskripsi gambaran protein spesifik urin dan berat molekul dan dilakukan uji beda terhadap kelas histopatologi dan aktivitasnya.Hasil didapatkan perbedaan gambaran pola protein spesifik urin dan jumlah peak pada pasien NL tidak aktif dan aktif. Berat molekul protein spesifik urin pada NL tidak aktif dan derajat ringan (kelas I,II) > 66kDa sedangkan pada NL aktif dan derajat berat (kelas III, IV, V) mempunyai protein urin tambahan dan protein berat molekul rendah 30-21kDa.Terdapat perbedaan gambaran jumlah peak dan berat molekul protein spesifik urin pada NL tidak aktif dan NL aktif. Pada NL derajat berat kelas III, IV, V dan NL aktif mempunyai protein urin tambahan dengan berat molekul rendah 30-21kD