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Abstrak :
Sel Punca Hematopoietik (SPH) memiliki potensi sebagai terapi regeneratif. Kriopreservasi umumnya dilakukan untuk menjaga kualitas SPH dari darah tali pusat. Larutan DMSO 10% adalah agen krioprotektan intraseluler standar dalam kriopreservasi SPH. Namun, DMSO bersifat toksik bagi sel pada suhu ruang dan pasien selama transplantasi. Oleh karena itu, konsentrasi DMSO perlu dikurangi dengan menambahkan krioprotektan ekstraseluler, seperti sukrosa atau madu sumbawa. Penelitian ini bertujuan untuk membandingkan kemampuan madu Sumbawa dan sukrosa sebagai krioprotektan ekstraseluler dalam melindungi SPH CD34+ selama kriopreservasi. Penelitian ini menggunakan desain eksperimental in vitro dengan tiga perlakuan medium kriopreservasi dan tujuh ulangan. Tiga perlakuan medium kriopreservasi terdiri atas DMSO 10% sebagai kontrol, DMSO 5% + madu Sumbawa 5%, dan DMSO 5% + sukrosa 5%. SPH CD34+ ditempatkan di Mr. Frosty dan disimpan dalam freezer pada suhu -80°C. SPH CD34+ dicairkan setelah 48 – 72 jam dan dilakukan analisis kualitas yang terdiri atas viabilitas, morfologi, dan stabilitas fenotipe. Hasil penelitian menunjukkan bahwa kombinasi krioprotektan DMSO 5% + madu sumbawa 5% berpengaruh positif dan memiliki perbedaan nyata (P<0,05) dibandingkan dengan DMSO 5% + sukrosa 5% terhadap viabilitas dan morfologi SPH. Namun, ratarata penurunan stabilitas fenotipe yang ditunjukkan dengan penurunan persentase CD34+ pada kontrol DMSO 10% (6,90 ± 8,60), DMSO 5% + sukrosa 5% (10,60 ± 9,20), dan DMSO 5% + madu sumbawa 5% (8,60 ± 11,50) tidak berbeda nyata (P>0,05). Kesimpulannya, kombinasi DMSO 5% + madu sumbawa 5% berpengaruh positif terhadap viabilitas dan morfologi HSC tetapi tidak terhadap stabilitas fenotipe. ......Hematopoietic Stem Cells (HSC) have potential as regenerative therapy. Cryopreservation was commonly practiced to preserve the quality of HSC from umbilical cord blood. DMSO 10% was standard intracellular cryoprotectant agents (CPAs) in HSC cryopreservation. However, DMSO is toxic to cells at room temperature and patients during transplantation. Therefore, the concentration of DMSO needs to be reduced by adding extracellular CPAs, such as sucrose or sumbawa honey. The objective of this study was to compare the ability of sumbawa honey and sucrose as extracellular CPAs to protect the HSC CD34+ during cryopreservation. The study used an experimental in vitro design with three treatments of cryopreservatin medium and seven replications. Three treatments of cryopreservatin medium consisted of DMSO 10% as a control, DMSO 5% + Sumbawa honey 5%, and DMSO 5% + sucrose 5%. HSC CD34+ in cryo medium was placed in Mr. Frosty and stored in the freezer at -80°C. HSC CD34+ were thawed after 48 – 72 hours and performed a quality analysis consisting of viability, morphology, and phenotype stability. The results showed that the cryoprotectant combination DMSO 5% + sumbawa honey 5% has positive effect and significant difference (P<0,05) compared with DMSO 5% + sukrosa 5% on the viability and morphology of HSC. However, the average decreasing phenotype stability as showed by decrease in percentage CD34+ in the DMSO 10% (6,90 ± 8,60), DMSO 5% + sukrosa 5% (10,60 ± 9,20), and DMSO 5% + madu 5% (8,60 ± 11,50) showed no significant difference (P>0,05). In conclusion, combination DMSO 5% + sumbawa honey 5% has positive effect on the viability and morphology of HSC but not on phenotype stability.

Abstrak :
Ruang lingkup dan Cara penelitian: Pengembangan metoda kontrasepsi pria Cara medikamentosa yang aman, efektif clan reversibel sekarang ini adalah penyuntikan intramuskular kombinasi hormon. Penyuntikan ini dapat menekan sekresi testosteron melalui penekanan gonadotropin hipofisis. Penyuntikan ini diharapkan tidak mempengaruhi fungsi hematopoietik, fungsi ginjal dan antigen spesifik prostat relawan yang turut berpartisipasi pada penelitian ini. Kombinasi hormon yang dipergunakan adalah kombinasi dosis rendah 100 mg TE + 100 mg DMPA dan kombinasi dosis tinggi 250 mg TE + 200 mg DMPA, disuntikkan setiap bulan dalam jangka waktu 12 bulan dan pemeriksaan fungsi hematopoietik, fungsi ginjal dan antigen spesifik prostat setiap 3 bulan. Penelitian ini dibagi dalam 3 We, yaitu fase kontrol atau pra-perlakuan (1 bulan), face penekanan (6 bulan) dan fase pemeliharaan (6 bulan). Pada fase kontrol atau pra-perlakuan dipilih 20 pria sehat dan subur yang memenuhi syarat pemeriksaan fisik dan laboratorium darah sebanyak 2 kali pemeriksaan normal, kemudian dibagi secara acak ke dalam 2 kelompok (masing masing kelompok 10 orang). Kelompok pertama mendapat penyuntikan kombinasi hormon dosis rendah dan kelompok kedua penyuntikan hormon kombinasi dosis tinggi. Parameter yang diteliti adalah: (a) fungsi hematopoietik, meliputi hematokrit, hemoglobin, leukosit, trombosit; (b) fungsi ginjal, meliputi ureum dan kreatinin darah; (c) antigen spesifik prostat. Hasil penelitian: Pemeriksaan laboratorium menunjukkan bahwa hasil kedua kelompok berada diantara batas normal: Ht. 41.67 - 47.46 %; Hb. 14.5 - 15.58 gldl; leukosit 7.48 - 11.54 (103/ul); trombosit 234.78 - 300.11 (103/ul); ureum 21.6 -- 28 mg/dl; kreatinin 0.92 - 1.21 mg/dl dan PSA 0.32 - 0.71 mg/dl. Setara keseluruhan penyuntikan hormon kombinasi dosis rendah 100 mg TE + 100 mg DMPA dan kombinasi dosis tinggi 250 mg TE + 200 mg DMPA tidak mempengaruhi fungsi hematopoietik, fungsi ginjal dan antigen spesifik prostat. Kesimpulan: Penyuntikan hormon kombinasi dosis rendah 100 mg TE + 100 mg DMPA dan kombinasi dosis tinggi 250 mg TE + 200 mg DMPA setiap bulan selama 12 bulan penelitian dan setiap 3 bulan pemeriksaan laboratorium tidak menimbulkan atau mengakibatkan perubahan bermakna pada fungsi hematopoietik, fungsi ginjal dan antigen spesifik prostat, sehingga kemungkinan aman sebagai slat kontrasepsi hormonal pria. ...... The Influence of Monthly Injection both a Low Dose and a High Dose Combination of TE + DMPA on the Hematopoietic and Kidney Functions and PSAScopes and methods of study: The medicinal approach to male contraception which is safe, effective and reversible is currently being investigated using a combination of hormones. The hormones, given by intramuscular injection, will suppress testosterone secretion through the suppression of gonadotropin release by the hypophysis. This study is carried out to investigate if there is any adverse effect on hematopoiesis (hematocrit, hemoglobin, leucocyte and thrombocyte as parameters), kidney functions (serum urea and creatinine), and prostate apecific antigen (serum) PSA during the use of this contraceptive means. Two hormonal combinations being evaluated are 1) a low dosage of 100 mg TE + 100 mg DMPA, and 2) a high dosage of 250 mg TE + 200 mg DMPA. The study is divided into 3 consecutive phases: control phase (1 month), suppression (6 months) and maintenance (6 months). The selected volunteers are twenty healthy and fertile males who show normal laboratory findings during the control period, which is carried out twice at a biweekly interval. They are then divided randomly into two groups of ten subjects each. Throughout the suppression and maintenance phases each member of the group receives a monthly injection of the low and high dosage hormonal combination, respectively. Venous blood samples are obtained every three months, the hematological and kidney parameters are examined at the Clinical Laboratory Department of the Cipto Mangunkusumo Hospital, and PSA measured by immunoassay (Abbott, IMx) at the Immunoendocrinology Laboratory of the Indonesia School of Medicine. The laboratory findings are analyzed by two-way anova, using a spreadsheet program (Lotus 123 or Exe1). Fidings and Conclusion: The laboratory parameters of the two groups are within the normal ranges throught out the study period: Ht. 41.67 - 47.46 %, Hb. 14.5 - 15.58 gldl, leucocyte 7.48 - 11.54 x 103/ul, thrombocyte 234.78 - 300.11 x 103/ul, ureum 21.6 - 28 mg/dL, creatinine 0.92 - 121 mg/dL and PSA 0.32 - 0.71 mg/dL. It is there for concluded that the administration of the combination of TE and DMPA, at both low and high dosages, has no adverse effect on hematopoiesis, kidney function and the prostate, and could therefor be considered safe for use in male contraception.

Abstrak :
Fenomena utama pada pasien MDS adalah sitopenia di darah tepi, namun disertai kondisi hiperselular di sumsum tulang. sCD40L dianggap sebagai sitokin yang dapat memicu sintesis TNFα sebagai sitokin proapoptosis dan memicu sintesis VEGF sebagai sitokin proangiogenesis pada MDS. Oleh sebab itu sCD40L dianggap berpotensi sebagai biopenanda untuk memperkirakan perburukan pada MDS. Penelitian ini bertujuan untuk membuktikan peran pajanan rh-sCD40L dalam menginduksi sintesis TNFα dan VEGF pada sel progenitor hematopoesis, serta membuktikan peran pajanan TNFα dalam memicu apoptosis pada sel progenitor hematopoesis dan sel mesenkim MDS. Penelitian ini merupakan penelitian eksperimental in vitro komparatif. Subjek penelitian adalah pasien MDS yang didiagnosis dan diklasifikasikan berdasarkan kriteria WHO 2008. Pada bone marrow mononuclear cells (BMMC) dipajankan dengan rh-sCD40L dan antiCD40L, kemudian dilakukan pemeriksaan ekspresi mRNA TNFα dan mRNA VEGF yang dikuantifikasi dengan qRT-PCR, serta pemeriksaan kadar TNFα dan VEGF yang diperiksa dengan metode ELISA. Pada sel CD34+, CD33+, CD41+, dan CD73+ dipajankan rhTNFα kemudian dilakukan pemeriksaan aktivitas kaspase-3 dengan imunoflowsitometri. Terdapat 15 sampel MDS terdiri dari 4 dengan diagnosis RCUD, 7 RCMD, dan 4 RAEB1, serta 7 sampel kontrol. Pajanan rh-sCD40L meningkatkan ekspresi mRNA TNFα secara bermakna dibandingkan pajanan antiCD40L. Pajanan rh-sCD40L meningkatkan kadar TNFα secara bermakna dibandingkan kontrol. Namun pajanan rh-sCD40L tidak meningkatkan mRNA VEGF dan kadar protein VEGF. Pajanan rhTNFα meningkatkan aktivitas kaspase-3 pada sel progenitor MDS terutama yang berdiferensiasi menjadi mieloid (CD33+) dan megakariosit-trombosit (CD41+). Pajanan rhTNFα meningkatkan aktivitas kaspase-3 pada sel mesenkim (CD73+) MDS Simpulan: sCD40L berperan dalam meningkatkan sintesis sitokin proapoptosis TNFα di level mRNA dan protein, namun tidak terbukti berperan dalam meningkatkan sintesis proangiogenesis VEGF. TNFα berperan dalam meningkatkan apoptosis terutama pada sel hematopoesis yang telah berdiferensiasi menjadi seri mieloid dan seri megakariosit-trombosit, dan berperan dalam meningkatkan apoptosis pada sel mesenkim.

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Cytopenia is the primary phenomenon in Myelodysplastic Syndrome (MDS) patients, amidst hypercellular bone marrow. The soluble CD40 ligand (sCD40L) is considered as a cytokine that can trigger synthesis of TNFα and VEGF. The former is known as a cytokine that promotes apoptosis while the latter promotes angiogenesis in MDS patients. Therefore, the sCD40L may serve as a potential biomarker to predict worsening of MDS. This study aims to prove the role of rh-sCD40L exposure in inducing the synthesis of TNFα and VEGF in hematopoietic progenitor cells, as well as to establish the role of TNFα exposure in triggering apoptotic activity in hematopoietic progenitor and mesenchymal cells of MDS. The study was a comparative in vitro experimental study. Subjects were MDS patients diagnosed and classified using the WHO 2008 criteria. Bone marrow mononuclear cells (BMMC) were exposed to rh-sCD40L and antiCD40L. The expressions of TNFα and VEGF mRNAs were then quantified by qRT-PCR, and the level of TNFα and VEGF were measured using the ELISA method. The CD34+, CD33+, CD41+, and CD73+ cells were exposed to rhTNFα, then the activity of enzyme caspase-3 was measured using the immunoflowcytometry. There were 7 control and 15 MDS samples with the following diagnoses: 4 RCUD, 7 RCMD, and 4 RAEB1. Compared to antiCD40L, it is found that exposure of rh-sCD40L significantly increased the expression of TNFα mRNA. The similar exposure also significantly increased the level of TNFα compared to controls. However, the exposure of rh-sCD40L did not increase the expression of VEGF mRNA as well as the level of VEGF. The exposure of rhTNFα was found to increase the activity of caspase-3 in MDS progenitor cells, particularly those differentiated into myeloid cells (CD33+) and megakaryocyte-thrombocyte cells (CD41+). The exposure of rhTNFα was found to increase the activity of caspase-3 in MDS mesenchymal (CD73+) cells. Conclusion: The sCD40L plays a role in increasing the synthesis of TNFα which favors apoptotic activity in mRNA and protein level, but not in improving the synthesis of VEGF that promotes angiogenesis. Furthermore, TNFα plays a role in increasing apoptotic activity of hematopoietic cells, particularly those that have differentiated into myeloid series and megakaryocyte-thrombocyte series cells. Also TNFα plays a role in increasing apoptotic activity of mesenchymal cells.

Abstrak :
This special issue on Notch regulation of the immune system summarizes recent advances and covers multiple aspects of Notch signaling within the hematopoietic and the immune system. This issue covers subjects including Notch function in embryonic and adult hematopoietic stem cells, lymphocyte development and function as well as in T cell leukemia.

Abstrak :
Sel Natural Killer (NK) adalah sel pelepas granul sitotoksik yang melisis patogen intracytoplasmic (yaitu infeksi virus atau bakteri) dan sel tumor/kanker sebagai bagian dari imunitas bawaan. Sel NK berasal dari diferensiasi sel punca hematopoietik (SPH) di jalur Common Lymphoid Progenitor (CLP). SPH diperoleh dari darah tali pusat, dengan jumlah SPH yang lebih tinggi daripada sumsum tulang. Berbagai protokol diferensiasi telah dilaporkan dengan jumlah sel NK dan fenotipe yang berbeda. Studi perbandingan efektivitas diferensiasi sel NK dengan sampel SPH yang dikultur dan SPH yang baru diisolasi masih minim. Penelitian ini bertujuan untuk membandingkan tahapan maturasi diferensiasi sel NK yang dihasilkan antara sampel SPH yang dikultur dan baru diisolasi menggunakan modifikasi protokol Dezell dkk. dengan menggunakan interleukin-2 (IL-2) tanpa keberadaan feeder cell. Kultur SPH dilakukan selama dua minggu sebelum diferensiasi untuk sampel SPH yang dikultur. Hasil kultur diferensiasi selama lima minggu dianalisis menggunakan flow cytometry untuk mengetahui keberadaan reseptor NKp46, pengamatan Giemsa untuk mengetahui tahapan maturasi sel NK, dan qRT-PCR untuk mengetahui ekspresi gen perforin dan granzyme B. Hasil penelitian menunjukkan bahwa sampel SPH yang dikultur menghasilkan jumlah sel NK di tahap dewasa (tahap 5) yang lebih tinggi dibandingkan sampel SPH yang diisolasi melalui pengamatan Giemsa. Hasil flow cytometry menunjukkan nilai MFI NKp46 yang berbeda signifikan pada kedua sampel, dengan keberadaan reseptor aktivasi NKp46 yang lebih tinggi dijumpai pada sampel isolasi di hari ke-35. Hal ini disebabkan oleh aktivitas ROS pada kultur SPH dan regulasi mikroRNA. Oleh karena itu, sampel SPH yang dikultur dan sampel SPH yang baru diisolasi mampu menghasilkan populasi sel NK yang dewasa. ......Natural Killer (NK) cells are cytotoxic-granule-releasing cells which lysis intracytoplasmic pathogens (ie. virus or bacteria infection) and tumor/ cancer cells as part of innate immunity. NK cells originate from differentiation of hematopoietic stem cells (HSCs) in the Common Lymphoid Progenitor (CLP) pathway. HSCs can be obtained from umbilical cord blood, with a higher number of HSCs than bone marrow. Various differentiation protocols have been reported with different NK cell yields and phenotypes obtained. Comparative studies on the effectiveness of NK cell differentiation with cultured HSC samples and freshly isolated HSC are still minimal. The aim of this study was to compare the different stages of NK cell differentiation maturation produced between cultured and newly isolated SPH samples using a modified protocol of Dezell et al. using interleukin-2 (IL-2) in the absence of feeder cells. For expanded HSC samples, cultures were carried out for two weeks before differentiation. The results of the differentiation culture for five weeks were then analyzed using flow cytometry to determine the presence of NKp46 receptors, Giemsa observations to determine the stages of NK cell maturation, and qRT-PCR to determine the expression of perforin and granzyme B genes. The results show that cultured HSC samples can produce a higher number of NK cells with a more mature stage than freshly isolated HSC samples by Giemsa's observations which showed the presence of NK cells at stages 5. The results of flow cytometry showed that the MFI NKp46 values ​​were significantly different in the two samples, with a higher NKp46 activation receptor found in the isolated samples on day 35. This is due to ROS activity on SPH culture and microRNA regulation. Therefore, the cultured HSC samples and freshly isolated HSC samples were able to produce mature NK cell populations.