Resistensi Mycobacterium tuberculosis terhadap beberapa obat anti tuberculosis pilihan utama dan pilihan kedua di Laboratorium Mikrobiologi FK UGM Tahun 2000 - 2004

Latar Belakang: Usaha penanggulangan tuberkulosis sering terhmbat oleh penyebaran strain Mycobacterium tuberculosa yang resisten multi obat. Salah satu upaya untuk mengatasi masalah tersebut adalah dengan penyampaian informasi mengenal data resistensi kuman secara berkala.Tujuan penelitian: Penelitian dilakukan dengan tujuan mengetahui pola resistensi Mycobacterium tuberculosa terhadap beberapa obat antituberkulosis, dalam rangka pemantauan strain bakteri resisten obat, terutama di Yogyakarta. Bahan dan care: Penelitian dilaksanakan sejak Januari 2000 sampai Desember 2004 dengan menggunakan 99 isolat Mycobacterium tuberculosa di Laboratorium Mikrobiologi, Fakultas Kedokteran UGM. Resistensi isolat-isolat tersebut terhadap obat pilihan pertama dan pilihan kedua dievaluasi. Suspensi bakteri dengan kepadatan setara dengan standar Mac Farland 1 (108 cfu/ml) ditanam pada medium Lowenstein Jensen yang mengandung masing-masing obat antituberkulosis (OAT) pilihan pertama yaitu: lsoniazid/INH (1,Oug/ ml), Sreptomisin (2,Oug/mI), Rifampisin (1,Oug/m1), Ethambutol (2,Oug/m1), dan obat pilihan kedua, yaitu: Kanamisin (lug/mi), Siprofloksasin (1ug/m1), Ofloksasin (5ug/mI). Sebagai kontrol suspensi bakteri ditanam di medium yang sama tanpa obat. Hasil: Pengamatan terhadap resistensi Mycobacterium tuberculosa menunjukkan bahwa sejumlah isolat telah resisten terhadap obat pilihan pertama yang diujikan, dengan kisaran 24,24% sampai 43,43%. Resistensi terendah adalah terhadap INH (24,24%) dan tertinggi Rifampisin (43,43%), sedangkan terhadap Streptomisin terdapat resistensi sebesar 33,33% dan terhadap Ethambutol 26,26%. Resistensi terhadap OAT pilihan kedua berkisar antara 14,29% sampai 49,50%. Resistensi tertinggi terhadap Kanamisin dan terendah terhadap Ofloksasin. Simpulan: Terhadap obat pilihan pertama, 74,75% dari isolat uji telah resisten terhadap satu OAT atau Iebih, 15,15% telah resisten terhadap semua OAT, dan hanya 25,25% isolat masih peka terhadap obat yang diujikan. Sementara itu 4.04% isolat merupakan strain resisten multi-obat (MDR-TB) karena resistan terhadap INH dan Rifampisin

The accuracy of fine needle aspiration biopsy to diagnose breast neoplasm

Breast lump is a very common complaint among women, especially during the reproductive year. Fine needle aspiration biopsy (FNAB) is a less invasive procedure. It is usually performed as an initial diagnosis prior to the operative procedure. The accuracy of the FNAB in Indonesia needs to be elaborated. The study aimed to evaluate the sensitivity and specificity of FNAB in diagnosing breast neoplasm. This is a retrospective study with cross sectional design, involving 145 patients with breast lump who underwent FNAB and histopathology examination in Dr. Sardjito General Hospital, Yogyakarta, from 2012 to 2014. Data analysis showed that female to male ratio was 23. 2:1 commonly occurred at 41-50 years old. Forty-one cases (28.28%) diagnosed as a benign lesion with fibrocystic changes as the most frequentcase (11.19%). The malignant case was 104 cases (71.72%) with ductal carcinoma as the highest case (51.49%). FNAB achieved a sensitivity of 85.58%, a specificity of 100% and a total accuracy of 89.66% in determining the benign or malignant breast lump. The accuracy, sensitivity and specificity of FNAB in diagnosing ductal carcinoma were 83.58%, 85.51% and 81.54%, respectively. The accuracy, sensitivity and specificity of FNAB to diagnose fibrocystic changes lesion were 85.82%, 26.67% and 93.28%, respectively. FNAB can be used as an alternative diagnostic tool to diagnose breast neoplasm. It provides rapid, cheaper, effective, valuable, and less invasive procedure in diagnosis of breast lump.

Malignant bilateral ovarian steroid cell tumor without androgenic manifestation: an unusual finding

Steroid cell tumor is a rarest ovarian neoplasm, classified as a pure stromal tumor and mostly is unilateral. Even though this tumor can exhibit malignant behavior but the morphology of cells showed benign characteristics which can become a diagnosis pitfall especially in the frozen section. Moreover patient without any hormonal imbalance or virilizing signs could make the diagnosis process more difficult. Here we reported a case bilateral steroid cell tumor of the ovary in a 42 y.o. unmarried woman without any virilization or hirsutism symptoms. Abdominal ultrasonography and computed tomography (CT) scan revealed a right ovarian solid tumor accompanied by ascites and right pleural effusion. There was significantly increased of Ca 125 level (1138 U/mL) and normal level of testosterone (0.10 ng/mL). Frozen section was done from the right ovarium mass and ascites fluid, the result was benign. From the total abdominal hysterectomy and bilateral salpingo-oophorectomy tissues,  histological picture showed diffuse and nests tumor separated by thin fibrous connective tissue with small round centered nuclei, mild atypia, and abundant pale cytoplasm. Large area of necrosis was found especially in the right ovarian tumor, tumor implant to the right fallopian tube and in the uterine serous layer. Periodic acid-Schiff (PAS) stain was negative in more than half tumor cells population. Immunostaining for Melan-A and Calretinin were focally positive, with Ki-67 labeling index ± 5%, and negative for cytokeratin 7 (CK7), cytokeratin 20 (CK20) and smooth muscle actin (SMA). Based on the tumor size, necrosis area, tumor implantation, and immunohistochemistry profiles, we conclude that were malignant steroid cell tumor. Currently, the patient is undergoing postoperative recovery and planned for platinum-based chemotherapy. A careful correlation between clinical and radiological findings, as well as histopathological results, is always essential, as is amply demonstrated by this particular case

Sinonasal or nasopharyngeal undifferentiated Carcinoma?: diagnostic pitfall and the role of Epstein-Barrvirus (EBV) and human papillomavirus(HPV) examination

Undifferentiated carcinoma of the head and neck is frequently observed in nasopharynx, however it may also occur in oropharynx, salivary gland and sinonasal. Overlapping lesions in those regionscreate difficulty in determining the origin of the tumor. Thus, it causes diagnostic pitfall not only for pathologists, but also for clinicians. A 40 yearold man, presented with nasal obstruction, epistaxis, diplopia, and headache for a yearand showed nasal cavitysinistra and nasopharynx masses on CT-scan. Lymph node enlargement was not detected. First biopsywas performed and histopathologically diagnosed as nasopharyngeal undifferentiated carcinoma(NPC), extended into nasal cavity. Chemo-radiation protocol for NPC was conducted, and showing uncomplete response. Second biopsy was done, and reviewed with the first biopsy result. Thetumourwas arranged insolid, syncytial and trabecular pattern, with vesicular nuclei, prominent nucleoli, and lack of lymphoplasmacytic infiltrat. Immunohistochemistry (IHC) analysis of p16, EBNA1 and LMP1 were negative. PCR analysis of HPV-18 was positive, while EBV detection showed negative result. General association of EBV with NPC suggests that the presence of latent EBV infection can serve as a positive marker for NPC. Therefore, in this case, the EBV negativity and strong HPV association led to diagnosis of SNUC. The distinction of sinonasal undifferentiated carcinoma (SNUC) or from NPC was important for appropriate management and therapy

Accuracy of fine needle aspiration biopsy to diagnose lymphadenopathy in Dr.Sardjito General Hospital, Yogyakarta, Indonesia

Lymphadenopathy is a non-specific enlargement of lymph nodes which may be caused by infection, cancer, or autoimmune disease. To date, only a few studies reported the diagnostic value of fine-needle aspiration biopsy (FNAB) in lymphadenopathy. This study was performed to evaluate diagnostic reliability of FNAB for benign and malignant lymphadenopathy. This was a retrospective cross-sectional study. The obtained data were statistically analyzed for its sensitivity, specificity, and accuracy. Out of 126 collected FNAB cases with histopathological confirmed results in Dr. Sardjito General Hospital, Yogyakarta, 85 (67.4%) were malignant lymphadenopathy, consisting of 42 metastatic tumor cases, 38 non-Hodgkin lymphoma (NHL) cases, and 4 Hodgkin lymphoma (HL) cases.The overall diagnostic sensitivity, specificity, and accuracy of FNAB in lymphadenopathy was 85.88, 70.73, and 80.95%, respectively. In diagnosing metastatic tumors, FNAB had sensitivity of 83.33%; specificity of 89.28%; and accuracy of 87.3%. The sensitivity, specificity, and accuracy of FNAB in diagnosing NHL was 60.52, 94.31, and 84.12%, respectively. FNAB had a sensitivity of 25%, specificity of 95,90%, and accuracy of 93.65% to diagnose HL. Meanwhile, the accuracy of FNAB in diagnosing malignancies in generalized lymphadenopathy, head-neck lymphadenopathy, and inguinal lymphadenopathy was 90.90; 81.39 and 44.44%, respectively. In conclusion,FNAB has moderate diagnostic value in diagnosing overall malignant lymphadenopathy, including metastatic tumors. FNAB also has some limitations in diagnosing NHL and HL, with sensitivity less than 70% for both diseases. However, it has high accuracy to diagnose generalized lymphadenopathy

Correlation between vascular endothelial growth factor (VEGF) expression with histopathological findings in osteosarcoma

Vascular endothelial growth factor (VEGF) expression is associated with malignancy progression, metastasis, and poor prognosis in many malignancies, including osteosarcoma. However, studies concerning correlations between VEGF expression and histopathological prognostic factors ofosteosarcoma are limited. This study aimed to evaluate the correlations between VEGF expression and histopathological findings in osteosarcoma’spatients.This was a cross-sectional study using formalin-fixed paraffin embedded (FFPE) samples of 32 osteosarcoma’s patients from Dr. Sardjito General Hospital, Yogyakarta. Histopathological findings of specimens were re-evaluated by two independent observers, recorded for the subtypes, invasiveness, grading, mitotic counts, and tumor infiltrating lymphocytes (TIL). Expression of VEGF was determined based on immunostaining and evaluated using immunoreactivity score (IRS).Chi-square and Spearman correlation test were used to analyze the association between variables. Range of VEGF expression score was 0 to 11, with mean 5.09. Significant negative correlation between the VEGF expression and TIL was observed (p=0.046). However, there was no significant correlations between the VEGF expression and osteosarcomas subtypes, invasion, grading or mitotic counts (p> 0.05). In conclusion, the VEGF expression is associated with TIL. Further study is needed to evaluate the roles of VEGF and lymphocytes in osteosarcoma development dan progression in order to better understand of the role of VEGF in immunotherapy of osteosarcoma

Categorization of 77 dystrophin exons into 5 groups by a decision tree using indexes of splicing regulatory factors as decision markers

<p>Abstract</p> <p>Background</p> <p>Duchenne muscular dystrophy, a fatal muscle-wasting disease, is characterized by dystrophin deficiency caused by mutations in the <it>dystrophin </it>gene. Skipping of a target <it>dystrophin </it>exon during splicing with antisense oligonucleotides is attracting much attention as the most plausible way to express dystrophin in DMD. Antisense oligonucleotides have been designed against splicing regulatory sequences such as splicing enhancer sequences of target exons. Recently, we reported that a chemical kinase inhibitor specifically enhances the skipping of mutated <it>dystrophin </it>exon 31, indicating the existence of exon-specific splicing regulatory systems. However, the basis for such individual regulatory systems is largely unknown. Here, we categorized the <it>dystrophin </it>exons in terms of their splicing regulatory factors.</p> <p>Results</p> <p>Using a computer-based machine learning system, we first constructed a decision tree separating 77 authentic from 14 known cryptic exons using 25 indexes of splicing regulatory factors as decision markers. We evaluated the classification accuracy of a novel cryptic exon (exon 11a) identified in this study. However, the tree mislabeled exon 11a as a true exon. Therefore, we re-constructed the decision tree to separate all 15 cryptic exons. The revised decision tree categorized the 77 authentic exons into five groups. Furthermore, all nine disease-associated novel exons were successfully categorized as exons, validating the decision tree. One group, consisting of 30 exons, was characterized by a high density of exonic splicing enhancer sequences. This suggests that AOs targeting splicing enhancer sequences would efficiently induce skipping of exons belonging to this group.</p> <p>Conclusions</p> <p>The decision tree categorized the 77 authentic exons into five groups. Our classification may help to establish the strategy for exon skipping therapy for Duchenne muscular dystrophy.</p

Mutation spectrum analysis of DMD gene in Indonesian Duchenne and Becker muscular dystrophy patients [version 3; peer review: 2 approved]

Background Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are allelic disorders caused by mutations in the DMD gene. The full mutation spectrum of the DMD gene in Indonesian patients is currently unknown. Mutation-specific therapies are currently being developed, such as exon skipping or stop codon read-through therapy. This study was conducted with the aim of identifying the mutation spectrum of the DMD gene in Indonesia to guide future development and application of feasible therapeutic strategies. Methods This study is a cross sectional study that enrolled 43 male patients with a clinical suspicion of DMD or BMD. Multiplex ligation-dependent probe amplification (MLPA) reaction was performed to screen for the common mutations in the DMD gene. Results Out of 43 subjects, deletions accounted for 69.77% (n=30) cases, while duplications were found in 11.63% (n=5) cases. One novel duplication spanning exons 2 to 62 was identified. Deletion mutations clustered around the distal (66.67%) and proximal (26.67%) hot spot regions of the DMD gene while duplication mutations were observed solely at the proximal region. Two false positive cases of single exon deletion detected through MLPA were attributed to sequence mutations affecting primer ligation sites, confirming the need to validate all single exon deletions when using this screening method. Analysis of available maternal DNA samples showed that the rate of de novo mutations (48.15%) appears higher than expected in this population. Out of 31 patients who were classified as DMD based on clinical and genotype characterizations, 60.47% (n=26) of cases were suitable for exon skipping therapy. Conclusion This is the first comprehensive study showing the feasibility of implementing the MLPA method for routine screening of DMD patients in Indonesia. This is also the first study showing the potential applicability of exon skipping therapy in the majority of DMD cases in the country