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Abstrak :
Latar belakang. Salah satu penentu manifestasi klinis dengue berat adalah kebocoran plasma. Limfosit dan monosit berperan dalam patogenesis kebocoran plasma infeksi dengue sehingga berpotensi sebagai prediktor kebocoran plasma. Tujuan. Menentukan kemampuan hitung jenis limfosit dan monosit demam hari kedua sebagai prediktor kebocoran plasma pada fase kritis infeksi dengue. Metode. Desain kohort retrospektif pasien rawat inap di RSUPN Cipto Mangunkusumo, RSUP Persahabatan dan RSPAD Gatot Soebroto dari tahun 2010 ̶ 2015, memenuhi kriteria inklusi: berusia > 15 tahun; didiagnosis dengue menurut WHO 1997; dikonfirmasi pemeriksaan NS-1/pemeriksaan serologis anti dengue; memiliki data darah perifer lengkap dan hitung jenis mulai demam hari ke-2; USG abdomen, dan/atau albumin pada fase kritis. Dilakukan analisis Reciever Operating Characteristic Curves (ROC curve) dengan interval kepercayaan (IK) 95% dan multivariat regresi logistik untuk memperoleh model prognostik. Hasil. Terdapat 63 subjek dianalisis. Insidens kebocoran plasma 49%. Nilai absolut limfosit dan nilai absolut monosit demam hari ke-2 berpotensi menjadi prediktor kebocoran plasma pada fase kritis dengan AUC 0,65 dan 0,64. Titik potong optimal nilai absolut limfosit dan nilai absolut monosit yang berpotensi sebagai prediktor kebocoran plasma sebesar 1323 dan 770. Nilai absolut limfosit memiliki sensitivitas 90%, spesifisitas 16%. Nilai absolut monosit memiliki sensitivitas 94%, spesifisitas 34%. Model prognostik nilai absolut monosit dan persentase limfosit meningkatkan AUC menjadi 0,723. Simpulan. Kemampuan prediksi kebocoran plasma nilai absolut limfosit dan nilai absolut monosit demam hari kedua lemah. Namun kemampuan tersebut ditingkatkan menjadi sedang oleh model prognostik yang melibatkan persentase limfosit dan nilai absolut monosit. ......Background. The severity of dengue infection was determined by plasma leakage. Lymphocytes and monocytes played an important role in the pathogenesis of plasma leakage in dengue infection so they potentially used as predictors for plasma leakage in a critical phase of dengue infection. Aim. Determined the percentage and absolute number of lymphocytes and monocytes measured on the second day of fever as a predictors for plasma leakage in a critical phase of dengue infection. Method. The research was retrospective cohort study of inpatients at Cipto Mangunkusumo Hospital, Persahabatan General Hospital and Gatot Subroto Military Hospital Jakarta from 2010 ̶ 2015. The inclusion criteria: age > 15 years, suffering from dengue infection according to the diagnostic criteria of WHO in 1997, confirmed by examination of NS-1 or serological anti-dengue, peripheral blood count and differential leucocyte count during treatment from second day of fever, abdominal ultrasound, and / or albumin in the critical phase. Analyses were performed using ROC curve. Multivariate analysis was performed to elicit prognostic models. Results. We determined 63 subjects. The incidence of plasma leakage was 49%. Absolute number of lymphocytes and monocytes on second day of fever were potentially useful as predictors for plasma leakage. The AUC was 0.65 and 0.644. The optimal cut-off point for absolute number of lymphocytes were 1323, the sensitivity was 90% and the specificity 16%. The cut-off for absolute number of monocytes was 770, the sensitivity was 94%, specificity 34%. We found optimal prognostic model which include percentage of lymphocytes and absolute number of monocytes. It could increase the AUC until 0,723. Conclusion. The absolute number of lymphocytes and monocytes on second day of fever in dengue infections were potentially useful as predictors for plasma leakage in a critical phase of dengue infection. Predictive capability could be increased by prognostic model which include percentage of lymphocytes and absolute number of monocytes as predictors.

Abstrak :
Latar Belakang: HER2 merupakan protoonkogen menjadi dasar pemberian terapi sel target pada adenokarsinoma gaster stadium lanjut. Penelitian hubungan antara gambaran klinis, endoskopi dan histopatologi dengan ekspresi HER2 masih menunjukkan hasil yang berbeda. Penelitian tentang HER2 sebagai prediktor kesintasan juga menunjukkan hasil yang berbeda. Penelitian ini bertujuan untuk mengetahui hubungan gambaran klinis, endoskopi, histopatologi dengan ekspresi HER2 dan hubungan antara ekspresi HER2 dengan kesintasan dua tahun. Metode: Penelitian kohort retrospektif pada subyek adenokarsinoma gaster yang baru terdiagnosis, berusia ³ 18 tahun di 4 rumah sakit di Jakarta, berobat dari 2015-2019, memenuhi kriteria inklusi: memiliki rekam medis yang lengkap, hasil pemeriksaan gastroskopi, blok parafin hasil biopsi. Slide biopsi diwarnai dengan pewarnaan imunohistokimia HER2 dan diinterpretasi dengan kriteria ToGA. Analisis statistik deskriptif dan bivariat dengan menggunakan chi square/tes fisher untuk menilai hubungan antara gambaran klinis, endoskopi, dan histopatologi dengan ekspresi HER2. Analisis kesintasan, bivariat dan multivariat dengan Cox-regresi menentukan pengaruh ekspresi HER2 terhadap kesintasan dua tahun. Hasil: Ekspresi HER2 positif ditemukan pada 12,3% subyek (15 dari 122 subyek). Ekspresi HER2 cenderung lebih tinggi pada metastasis ke hati, klasifikasi Borrman tipe I/II, diferensiasi baik/sedang, tipe intestinal berdasarkan Klasifikasi Lauren memiliki dengan proporsi masing-masing: 17.6% RR(IK95%)=1.726 (0.665-4.480), 16.1% RR(IK95%)=1,768(0,670-4,662), 14.3% RR(IK95%)=1,304(0,505-3,363), 13.9% RR(IK95%)=1,389(0,505-3,817).Ekspresi HER2 positif tidak berhubungan dengan kesintasan dua tahun, HR (IK95%)=1,12(0,609-2,058). Simpulan: Matastasis hati, klasifikasi Borrman, letak tumor, diferensiasi tumor dan klasifikasi Lauren tidak berhubungan bermakna secara statistik terhadap ekspresi HER2. Ekspresi HER2 positif tidak berhubungan dengan kesintasan dua tahun pada adenokarsinoma gaster. ......Background: HER2 is a proto-oncogene which important for administering of target cell therapy in advanced gastric adenocarcinoma. Research on clinical, endoscopic, and histopathological features shown conflicting in association with HER2 expression. Studies on HER2 as a predictor of survival still show different results. This study aims to determine association of the clinical, endoscopic, and histopathological features with HER2 expression and association of HER2 expression with 2-year survival. Methods: A retrospective cohort study on newly diagnosed gastric adenocarcinoma subjects, aged 18 years old at 4 hospitals in Jakarta, receiving treatment from 2015-2019, meeting the inclusion criteria: having complete medical record, results of gastroscopy examination, and paraffin block tumor biopsy results. The biopsy slides were stained with HER2 immunohistochemical staining and interpreted according to the ToGA criteria. Descriptive and bivariate analysis by using chi-square or fisher's test assessed the relationship between clinical, endoscopic, and histopathological features with HER2 expression. Survival, bivariate and multivariate analysis with cox regression method were used to determine the effect of HER2 expression on 2-year survival. Results: Positive HER2 expression was found in 12.3% of subjects (15 of 122 subjects). HER2 expression tends to be higher in metastases to the liver, Borrman classification type I/II, good/moderate differentiation, intestinal type based on Lauren's classification has the respective proportions:17.6% RR (95%CI)=1.726 (0.665-4.480), 16.1% RR (95%CI)=1,768 (0,670-4,662), 14.3% RR (95%CI)=1,304 (0,505-3,363), 13.9% RR (95%CI)=1,389 (0,505-3,817). Positive HER2 expression was not associated with 2-year survival with HR (95%CI) =1.12 (0.609-2.058).

Abstrak :
Penggunaan antibodi poliklonal dalam sis tern pondeteksi antigen P24 HIV- 1 layak untuk dipertimbangkan mengingat variasi susunan epitop P24 pada berbagai subtipe HIV -I berpotensi mengaldbatksn kegagalan pengenalan epitop oleh antibndi monoklonal. Antibodi poliklonal yang dipero!eh melalui induksi dengan antigen rekombinan berpotensi bereaksi seeara non spesifik terhadap protein kontaminan yang terdapat dalam sediaan antigen rekombinan sehingga dapat berpengaruh pada spesifisitas sistem pendeteksi antigen. Pada penelitian sebelumnya diperoleh informasi mengenai reaksi non spesifik serum anti P24 HIV -I poliklonal yang dihasilkan melalui imunisasi kelinci, khnsusnya terhadap antigen E.coli dan Bovine Serum Albumin (BSA). Oleh ksrena penelitian ini, maka diteliti efek purifikssi dengan kromatografi afinitas dalam menghilangkan reaktivitas non spesifik antara serum anti P24 dengan E.coli dan BSA. Dampak ini dinilai dengan menggunakan teknik sandwich Enzyme Linked Immunoassay (ELISA). Purifikasi dilaknkan dengan menggunakan dua kolom kromatografi afinitas dengan ligan E.coli pada kolom pertama dan ligan P24 rekombinan HIV-1 pada kolom kedua CnBr -.sepharose digunakan sebagai matriks. Proses elusi menggunakan glycine HCI, pH 2,7. Eluen basil purifikasi dikonfirmasi dengan teknik SDS PAGE, western blot dan sandwich ELISA pendeteksi antigen P24 HIV -I. Pada SDS PAGE terbentuk pita an tara berat molekul 45-116 kDa yang menunjukkan pita antibodi. Pada uji western blot terdapat pita spesifik protein P24 rekombinan H!V -! dan tidak muncul pita non spesiflk terhadnp E.ooli. Sedangkan pada uji sandwich ELISA, eluen menunjukkan nilai abwrbansi yang lebih tinggi dibandingkan dengan kontrol negatif, dan terdapat penurunan nilai absorbansi dibandingkan dengan sistem laripa antibodi yang dipurifikasi. Nilai absorbansi cluen juga tidak menunjukkan hasil yang berbeda dengan kontrol negatif jika direaksikan dengan antigen E.coli. Namun reaktifitas non spesiflk e]uen dengan BSA pacta sistem sandwich ELISA tidak berbeda dengan reaktifitas serum prepuriflkasi. Pada uji western blot dan sandwich ELISA diperoleh pula informasi yang menunjukkan adanya reaksi non spesifik antara antibedi anti-kelinci yang digunakan dengan protein E.eo!L Puriflkasi antibedi dengan menggunakan metode kromatografi afmitas telah berhasil dilaknkan dengan kondisi optimal pemurnian dan telah diperoleh eluen yang mengandung antibodi terhadap P24 rekombinan HIV -1 yang tidak bereaksi dengan protein E.coli Reagensia yang digunakan dalam sistem pendeteksi berpotensi menimbulkan ikatan non spesiflk yang dapat mengganggu nilai absorbansi sehingga sebelum digunakan harus dinilai kelayakannya untuk digunakan dalam sistem diagnostik tertentu.

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Polyclonal antibody may be important to be considered in sandwich ELISA which detected HIV-1 P24 due tO the existing variations in P24 epitope structure. Variations in epitope structure has the potential to produce false negative result in serologic diagnostic system that utilize monoclonal antibody. Polyclonal antibody induced by recombinant antigen could exhibit non specific reaction due to the presence of coniaminanl in antigen preparation that originates from the host used for expression of the recombinant antigen. In the previous research, we have already known that our polycional antibody in P24 H!V recombinant immunized rabbit serum had non specific reaction with E. coli protein and Bovine Serum Albumin (BSA) in sandwich ELISA system for detecting HIV-1 P24 recombinant protein. This was the ma}or reason to purifY the antibody with affinity chromatography. Our goal war to reduce the non specific reaction. We used sandwich Enzyme Linked Immunoassay (ELISA) to see the effect of purification. We used two columns affinity chromatography with different ligand in each column and CnBr sepharose as solid support. The first column utilized E.coli protein as ligand and the second one used HIV-1 P24 recombinant protein. We used glycine HCI, pH 2, 7 to elute antibody from qfflnity chromatography column. Eluen from the second column was coNfirmed with SDS PAGE, western blot and sandwich ELISA. SDS PAGE followed by comassie blue staining showed specific bands between 45-116 kDa molecular weight, which were interpreted as heavy and light chain fragments of antibody. Purified antibody in the second eluen was shown to be reactive with HIV-1 P24 recombinant protein but not E. coli protein by western blot analysis. There was a decline in the absorbance when eluen was used as detection antibody in sandwich ELISA system, compared with the system that utilized pre purified antibody. We also observed there was non specific reaction between the components in sandwich ELISA for detection of H/V-1 P24 recombinant antigen, in that we found the antibody against anti- rabbit JgG which was used in sandwich ELISA system had non specific reaction with E,cali protein. We concluded that we gained optimal condition in polyclonal antibody purification to reduce non specific reaction between polyclonal antibody to HW-1 P24 recombinant antigen with E.coli protein. Reagents which used in our sandwich ELISA system potentially Caused non specific reaction so we have to consider their application.