The Association of FLT3-ITD Gene Mutation with Bone Marrow Blast Cell Count, CD34, Cyclin D1, Bcl-xL and hENT1 Expression in Acute Myeloid Leukemia Patients

marker for risk classification in acute myeloid leukemia (AML) therapy. In this study we aimed to investigate the association of FLT3-ITD gene mutation with bone marrow blast cell count, CD34 expression as malignant cell burden, cyclin D1 and Bcl-xL expressions as indexes of cell proliferation and anti-apoptosis and human equilibrative nucleoside transporter 1 (hENT1) expression as cytarabine transporter during AML treatment. Methods: We investigated FLT3-ITD mutations, bone marrow blast cell count, CD34, cyclin D1, Bcl-xL and hENT1 expression in bone marrow aspirates from 22 de novo AML patients in a cross sectional study. Results: FLT3-ITD mutations were observed in 5 out of 22 de novo AML patients (22.7%). Patient with FLT3-ITD mutations had higher blast cell counts (79.5% vs 56.1%, P=0.004). In patients with FLT3-ITD mutations, CD34 and cyclin D1 expressions were higher (MFI 328.80 vs 25.78, P=0.003 and MFI 74.51 vs 57.15 P=0.005) than the patients without mutations. hENT1 expression in AML with FLT3-ITD mutation was lower (MFI 29.64 versus 56.32, P=0.0000) than in mutation-free AML. There was no significant difference in Bcl-xL expression between patients with and without mutations (P=0.61). Conclusion: A significant association was found between FLT3-ITD gene mutations in AML patients with bone marrow blast cell count, CD34, cyclin D1 and hENT1 expressions, however no association was obtained with Bcl-xL expression. These findings support the role of such mutation in pathogenesis of AMLand its contribution in rearrangement of standard therapy with cytarabine in management of AM

Sensorineural Hearing Loss in Juvenile CML:A Rare Case Report in Surabaya, Indonesia

Abstract: Chronic myeloid leukemia (CML) is a type of myeloproliferative neoplasm (MPN) in which granulocytes are the primary proliferating cells. CML in pediatric patients (juvenile CML) is an uncommon MPN, representing 2–3% of children newly diagnosed with leukemia. Sensorineural hearing loss that progresses rapidly is an uncommon early sign of a hematological disorder in patients with CML. This article presents the case of a 10-year-old patient with complaints of bilateral hearing loss for 2 weeks. Pure-tone audiometry indicated sensorineural hearing loss of the left ear and mixed hearing loss of the right ear. After an in-depth evaluation of a bone marrow smear and positivity for the BCR-ABL fusion gene, it was concluded that the patient had chronic-phase juvenile CML. However, hearing loss recovery after hydroxyurea therapy could not be observed because the patient died after suffering an uncontrolled seizure on day 14 of hospitalization

PERBANDINGAN NILAI DIAGNOSTIK HDL DENGAN RASIO KOLESTEROL TOTAL : HDL PADA PASIEN PJK

Latar belakang:Penyakit Jantung koroner adalah penyakit yang menyerang banyak penduduk di berbagai negara termasuk Indonesia. WHO pada tahun 2013 memperkirakan, penderita yang meninggal akibat PJK (Penyakit Jantung Koroner) akan meningkat sampai sebesar 23,3 juta jiwa pada tahun 2030. Di Indonesia sendiri, proporsi penderita PJK semakin meningkat dari tahun ke tahun. Terdapat beberapa faktor resiko yang menjadi penyebab dari penyakit PJK. Beberapa diantaranya adalah kadar kolesterol darah yang tinggi atau dikenal sebagai hiperkolesterolemia.Banyaknya jenis dari kolesterol seperti kolesterol total, HDL, LDL, rasio kolesterol total / HDL, dan lain sebagainya dapat digunakan sebagai indikator daam mendeteksi PJK. Pada penelitian ini peneliti akan mendeskripsikan apakah terdapat perbedaan antara penggunaan rasio kolesterol total / HDL dengan HDL saja pada pasien penderita PJK di RSUD Dr Soetomo Surabaya. Tujuan: menggambarkan profil lipid HDL dan rasio kolesterol total / HDL pada penderita PJK di RSUD Dr. Soetomo Surabaya, dan mengetahui ada tidaknya perbedaan antara nilai prediktif antara rasio kolesterol total/HDL dengan HDL saja. Metode: penelitian ini menggunakan metode Analitik retrospektif dengan menggunakan metode total sampling dari penderita penyakit jantung koroner di RSU Dr Soetomo Surabaya Bulan Januari-Maret 2014 dan dilakukan uji sign test untuk mengetahui signifikansinya dengan kriteria α: 0,05 dan CI sebesar 95%. Hasil: Dari penelitian ini dapat disimpulkan bahwa terdapat perbedaan yang bermakna antara rasio kolesterol total : HDL dengan HDL saja sebagai nilai prediktif dalam pendeteksi penyakit PJK (P = 0.04)

Diagnostic Value of Myeloperoxidase Index in Bacterial Infections

Infectious diseases remain a serious problem in Indonesia. Myeloperoxidase (MPO) is a substance released by neutrophils, which activates the synthesis of hypochlorous acid (HOCL) from hydrogen peroxide (H2O2) and chloride ion (Cl-). Hypochlorous acid plays a vital role in the body's defense against infection. Myeloperoxidase Index (MPXI) is a parameter in the hematology analyzer Advia 2120i based on the principle of flow cytometry. This study aimed to determine the diagnostic value of MPXI in patients with bacterial infections. The study was a cross-sectional observational analysis. The samples consisted of a group of patients with bacterial infection and a group of healthy subjects. The specimens used in this study were whole blood + anticoagulant (EDTA) in a purple tube with a volume of 3 mL to determine the MPXI value in both groups using ADVIA 2120i hematology analyzer. The study subjects consisted of a group of patients with bacterial infections (69 patients) and a group of healthy subjects (33 people). Analysis of the MPXI ROC curve with a cut-off ≥ -5.8 and < -5.8, showed AUC of 0.323 (CI=95%, p=0.004), sensitivity of 34.8%, specificity of 39.4%, Positive Predictive Value (PPV) of 54.5%, and Negative Predictive Value (NPV) of 22.4%. Due to its low diagnostic value, the MPXI value was not recommended to be used as a diagnostic instrument for bacterial infections. Also, further research was highly needed

Difference Expressions CD34 in Acute Myeloid Leukemia Cell Culture in the Administration of Cytarabine-Daunorubicine Dose Standards

The cure rate for patients with Acute Myeloid Leukemia (AML) is 20-75%. Standard-dose cytarabine + (SDAC)-daunorubicine gives a remission rate of ± 60%, and the case of relapse is frequently found. In-vivo CD34 expression is a reliable and straightforward test that must evaluate AML patients' response to predict the response of chemotherapy + induction phase accurately. Differences in in-vitro CD34 expression are expected to be able to predict chemosensitivity in AML patients. An experimental post-test-only control group study was conducted from May to December 2019, and 8 AML subjects were found. Peripheral Blood Mononuclear Cells (PBMC) were isolated from peripheral blood samples of patients with AML collected in EDTA tubes. The PBMC isolated from peripheral blood were divided into two groups, and each group contained 106 PBMC cells in culture media. The control group (without treatment) and the SDAC-daunorubicine group were 0 + incubated for 4 hours at 37 C with a 5% CO2 atmosphere. The expression of CD34 was measured using FACSCalibur™, while + CD34+ percentage was calculated with CellQuest™ software. The percentage of CD34 in the control, SDAC + DNR, showed a significant difference with p < 0.001. This study showed a significant difference between the control group and the group + administered with the standard dose of cytarabine-daunorubicine with p < 0.001. The average CD34 expression in the + SDAC-DNR treatment group was higher than in the control group. CD34 markers cannot be used as predictors of chemosensitivity in the administration of chemotherapy

Thymoquinone modifies CD4+:CD8+ ratio without affecting tumor necrosis factor‑α and interleukin‑1β levels in Wallerian degeneration crush injury rat model

Wallerian degeneration following nerve injury not only suppresses CD4+ T-cell responses but also pro- motes pro-inflammatory immunological responses through TNF-α and IL-1β. Recent research suggests that thymoquinone might enhance nerve recovery by exerting anti-inflammatory effects on both the innate and adaptive immune systems. This study aims to evaluate the effect of thymoquinone on neuroinflammation in a sciatic nerve crush injury, as represented by TNF-α, IL-1β, and the CD4+:CD8+ ratio. In this study, 126 crush injury Wistar rats were divided into three main groups: placebo, thymoquinone 100 mg/kg, and thymoquinone 250 mg/kg administered daily. Rats were euthanized at six distinct time points: 12, 18, and 24 hours, as well as on day-5, day-6, and day-7. TNF-α and IL-1β levels were assessed using the Enzyme- Linked Immunosorbent Assay (ELISA). The CD4+:CD8+ ratio in peripheral blood was determined via flow cytometry. No significant TNF-α differences was found between treatment and placebo groups. However, on day 6, IL-1β was significantly lower in the TQ 250mg/kg group than in the placebo (p=0.008). A similar but non-significant trend existed on days 6 and 7. On day 5, both TQ groups showed a higher, statistically significant CD4+:CD8+ ratio compared to placebo (p=0.007), a trend that continued to day 7 but not sta- tistically significant. Daily TQ administration did not consistently reduce TNF-α and IL-1ß levels. However, both doses elevated the CD4+:CD8+ ratio during the early stages of Wallerian degeneration, suggesting a potential benefit of TQ on nerve regeneration

Platelet-rich fibrin (PRF) graft and amniotic membrane graft on transforming growth factor-β (TGF-β) and type 1 collagen post conjunctival excision

Abstract Background: Conjunctiva, when injured, can heal spontaneously. However, in extensive wounds, wound healing is often accompanied by scarring and wound contraction. Currently, for cases of disease and trauma to the surface of the eyeball, it can be reconstructed using the amniotic membrane, but there are still weaknesses in the use of amnion. Platelet-rich fibrin (PRF) is currently being developed for the reconstruction of ocular surface wounds. Methods: A total of 20 male New Zealand rabbits (20 eyes) were divided into two groups. In the first group, conjunctival excision and PRF membrane graft suturing were performed on the right eye conjunctival defect, while the second group had an amniotic membrane sutured on the right eye conjunctival defect. On the 14th day, enucleation was carried out and continued with histopathological examination using immunohistochemistry using TGF antibodies and type 1 collagen to evaluate the levels of TGF and type 1 collagen in the conjunctiva. Results: The results after 14 days of treatment showed that there was a very significant difference in TGF Beta expression between the AMT group and the PRF group (p = 0.007*; 0.05). Conclusion: There were differences in the expression of TGF-β and type 1 collagen in the administration of the PRF membrane compared to the administration of the amniotic membrane after conjunctival excision of experimental male New Zealand rabbits on day 14

Thymoquinone Modulates Local MMP-9, IL-10, and IgG in Sciatic Nerve Crush Injury Animal Model

BACKGROUND: Interleukin (IL)-10 is involved in Wallerian degeneration after peripheral nerve crush injury. Oral thymoquinone was previously observed to decrease local immunoglobulin-G (IgG) in a crush- injured rat model. No study has evaluated the pathway of various thymoquinone dosages on local IgG and IL-10 in this injury. METHODS: This experimental study used 126 Rattus norvegicus Wistar rats that were divided into 18 groups: six groups received a placebo, the other six groups received thymoquinone at 100 mg/kg/day and the last six groups received thymoquinone at 250 mg/kg/day, respectively. Rats were sacrificed at 12, 18, 24, 5x24, 6x24, and 7x24 hours. Matrix metalloproteinase-9 (MMP-9), IL-10, and local IgG levels were assessed by Enzyme-Linked Immunosorbent Assay (ELISA). The nuclear factor KappaB (NF-κB) expressions on Schwann cells were examined by flow cytometry. Path analysis was performed using SmartPLS. RESULTS: The path analysis showed that 100mg/kg/day of thymoquinone significantly decreased NF-κB expression. However, NF-κB did not affect local MMP-9, and MMP- 9 had no significant relationship with local IL-10 and IgG. Thymoquinone 250 mg/kg/day also significantly inhibited NF-kB expression, decreased local MMP-9, and, in turn, decreased local IL-10 and IgG. CONCLUSION: Administration of oral thymoquinone 250 mg/kg/day decreases local IgG and IL-10 levels via suppressing NF-κB expression and MMP-9 level

Analisis Hubungan Status Mutasi Gen Flt3-Itd, - Tkd (D835) Dengan Jumlah Blast, Status Ekspresi Cd34, Ekspresi Bcl-Xl, Cyclin D1 Dan Hent1 Pada Penderita Leukemia Mieloid Akut

Latar belakang : Mutasi gen FMS like tyrosine kinase 3 – internal tandem duplication, tyrosine kinase domain (FLT3-ITD, -TKD(D835)) berhubungan dengan prognosis yang buruk, tingginya kekambuhan, rendahnya ketahanan hidup keseluruhan pada penderita LMA. Hubungan mutasi ini dengan aktivitas proliferasi sel blast, aktivitas anti-apoptosis, dan transporter transmembran obat ara-C penting diteliti lebih lanjut. Tujuan : Analisis hubungan status mutasi gen FLT3-ITD, -TKD(D835) dengan jumlah sel blast, status ekspresi CD34, ekspresi cyclin D1, Bcl-xL, human ezuilibrative nucleoside transporter 1 (hENT1) pada penderita LMA. Metode : Penelitian potong lintang dengan 35 penderita LMA de novo dilakukan pemeriksaan mutasi gen FLT3-ITD, -TKD(D835), jumlah blast, status ekspresi CD34, ekspresi cyclin D1, Bcl-xL, dan hENT1 Hasil : Sebanyak 8 dari 35 penderita LMA (22,9%) ditemukan adanya mutasi FLT3-ITD, dan tidak ditemukan penderita dengan mutasi FLT3-TKD(D835). Penderita LMA dengan mutasi FLT3-ITD memiliki jumlah blast lebih tinggi (71,3% vs51,5%, p=0,002), ekspresi cyclin D1 lebih tinggi (median fluorescent index / MFI 68,77 vs 55,44, p = 0,036) dan ekspresi hENT1 lebih rendah ( MFI 30,6 vs 58,73, p = 0,0001) daripada penderita tanpa mutasi. Penderita LMA dengan dan tanpa mutasi FLT3-ITD tidak memiliki perbedaan status ekspresi CD34 dan ekspresi Bcl-xL (p=0,97, p=0,38). Mutasi gen FLT3-ITD memiliki hubungan dengan jumlah blast (r=0,55, p=0,002), ekspresi hENT1 (r=0,7, p=0,0001) dan ekspresi cyclin D1 (r=0,34, p=0,048). Mutasi tersebut tidak berhubungan dengan status ekspresi CD34 dan ekspresi Bcl-xL. Kesimpulan : Mutasi gen FLT3-ITD pada LMA berhubungan dengan jumlah blast, ekspresi cyclin D1 dan hENT1. Mutasi tersebut tidak berhubungan dengan ekspresi Bcl-xL dan status ekspresi CD34

Detection of FLT3 Gene Mutations In Patients With Acute Myeloid Leukemia In Surabaya, Indonesia: A Single-Center Study

acute myeloid leukemia (AML). Almost 67% of patients with AML with FLT3 gene mutation cannot reach complete remission after induction therapy, and they are also at high risk of relapse. We aimed to investigate the FLT3-ITD, -TKD (D835) gene mutation prevalence in patients with AML at Surabaya and their association with leukocyte and bone marrow blast cell count. Methods: 20 de novo patients with AML were recruited during February–July 2018. They were investigated through routine AML check-up and detection for FLT3-ITD, -TKD (D835) gene mutation. Results: Four patients with de novo AML (20%) had FLT3-ITD gene mutation, and one patient had FLT3-TKD (D835) gene mutation. Median leukocyte count in patients with AML with FLT3-ITD mutation was higher than wild type patients ( 146.3×103/ μL vs 16.4 x 103 / μL, P=0,002). The mean bone marrow blast cell count was higher in patients with AML with FLT3-ITD mutation than wild type patients ( 86.5% vs 57.9%, P=0,047). The difference in leukocyte and bone marrow blast cell count in patients with FLT3-TKD (D835) mutation could not be analyzed since there was only one patient with this mutation. Conclusion: The prevalence of FLT3-ITD gene mutation in patients in AML in Surabaya was 20% and FLT3-TKD (D835) was 5%. Patients with FLT3-ITD gene mutation was associated with leukocyte and bone marrow blast cell count