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Pengaruh Perbedaan Metode Ekstraksi Metabolit Sekunder Streptomyces sp. GMR22 terhadap Toksisitas pada Sel BHK-21 Mentari, Diani; Naima, Mirtani; Wulansari, Riska; Widada, Jaka; Nuringtyas, Tri Rini; Wibawa, Tri; Wijayanti, Nastiti
Pharmacon: Jurnal Farmasi Indonesia Vol 16, No 1 (2019)
Publisher : Universitas Muhammadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/pharmacon.v16i1.8032

Streptomyces sp. GMR22 is local isolate from Wanagama 1 Forest in Yogyakarta. They have the potential to be developed to produce active compounds because have PKS and NRPS genes.The active compounds from isolation are strongly influenced by various factors, one of them is extraction techniques. Effect difference of extraction technique will be affected by the quality of secondary metabolites produced.The purpose of this study was to compare the cytotoxicity effects of secondary metabolites of Streptomyces sp. GMR22 which have extracted with different stages from previous studies. The extraction technique was carried out by multilevel separatory funnel extraction methods, which was first extracted using non-polar solvent (n-hexane) and then extracted using semi-polar solvent (ethyl acetate). This research is important because in previous studies (separatory funnel only extracted using ethyl acetate) with the use of the lowest concentration in the dengue virus antiviral test (further test) caused 100% of deaths in BHK-21 cells.This study indicate that multilevel extraction result in lower CC50 value than previous studies. There are 49.160 µl/ml (n-hexane extract) and 284.56 µl/ml (ethyl acetate extract) while water extract is 464,38 µl/ml. FTIR compound analysis show that the three extracts produced have different spectrum patterns, especially in the n-hexane and ethyl acetate extract. Value of CC50 is not too high, it is expected that the secondary metabolites contained in the extracts can be used for further analysis such as antiviral testing because it is safe for normal host cells such as BHK-21 cells

PENGARUH WAKTU SIMPAN TERHADAP PERUBAHAN PH, GLUKOSA, LDH, KALSIUM, MVP SEBAGAI INDIKATOR KUALITAS KOMPONEN DARAH THROMBOCYTE CONCENTRATE Mentari, Diani; Pebrina, Relita; Nurpratami, Diah
Biomedika Vol 12, No 1 (2020): Biomedika Februari 2020
Publisher : Universitas Muhamadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/biomedika.v12i1.8981

ABSTRAK Thrombocyte Concentrate (TC) merupakan komponen darah yang ditranfusikan untuk pasien yang mengalami pendarahan, kelainan fungsi trombosit dan trombositopenia. Pada beberapa literatur menyebutkan bahwa sediaan TC secara in vitro dapat disimpan selama 5-7 hari. Quality Control sebelum TC ditransfusikan yaitu melalui pengamatan secara visual ada tidaknya swirling dan melihat tingkat kejernihan TC. Namun pengamatan bersifat subjektif sehingga belum terstandar. Penelitian ini bertujuan untuk melihat pengaruh waktu penyimpanan TC terhadap kualitasnya. Kualitas TC diukur melalui beberapa cara yaitu pengukuran pH, kadar glukosa, Laktat Dehidrogenase (LDH), kalsium serta profil darah yang meliputi jumlah trombosit dan Mean Platelet Volume (MPV). Analisis kadar glukosa, LDH, dan kalsium dilakukan secara kolorimetri menggunakan spektrofotomter sedangkan profil darah diukur menggunakan hematology analyzer. Hasil menunjukkan bahwa TC yang disimpan pada hari ke-9 terjadi penurunan pH sebesar 7,38%, glukosa 20,10%, LDH 42,89%dan kalsium 62,54%. Jumlah trombosit mengalami penurunan sebesar 24,41% dan MVP mengalami kenaikan 18,84%. Kesimpulan pada sampel TC yang masih terdapat swirling, namun terjadi penurunan kualitas TC yang ditandai dengan penurunan jumlah trombosit serta kenaikan nilai MVP. Selain itu semakin lama waktu penyimpanan akan menurunkan kadar pH, kadar glukosa, LDH, dan kalsium. Kata Kunci: Masa Simpan Trombosit, pH, Glukosa, Kalsium, MPV. ABSTRACT Thrombocyte Concentrate (TC) is the blood component tranfused for the patients with bleeding, abnormalities of platelet function and thrombocytopenia. In some literatures, it is mentioned that the TC preparation in vitro can be stored within 5-7 days. Quality Control before TC is transfused through a visual observation on the presence or absence of swirling and the clarity level of TC. However, this observation is subjective and makes it unstandardized. This research aims to see the effects of the shelf life of thrombocytes on its quality. The TC quality was measured through a number of ways including: pH measurement, glucose level, Lactate Dehydrogenase (LDH), calcium and profiles of blood including number of thrombocytes, and Mean Platelet Volume (MPV). The analysis on the glucose level, LDH, and calcium was conducted by colorimetrical manner using the spectrophotometer and blood profile was measured using hematology analyzer. The result showed that TC stored in day 9 experienced the decrease of pH at 7.38%, glucose at 20.10%, LDH at 42,89% and calcium at 62.54%. The number of thrombocytes experienced a decrease of 24.41% and MVP experienced an increase of 18.84%. In conclusion, the sample of TC that had swirling experienced a decrease in the TC quality as characterized with the decrease of number of thrombocytes and the increase of MVP value. In addition, the longer storage could decrease the level of pH, level of glucose, LDH, and calcium. Keywords: Platelet Storage, pH, Glucose, Calcium, MPV.

Perbedaan Waktu Penambahan Reagen AHG Berpengaruh Terhadap Hasil Pemeriksaan Uji Silang Serasi Metode Tabung Syaqina Rassajati; Diani Mentari; Relita Pebrina; Hieronymus Rayi Prasetya
Jurnal Analis Medika Biosains (JAMBS) Vol 9, No 1 (2022): JURNAL ANALIS MEDIKA BIOSAINS (JAMBS)
Publisher : Poltekkes Kemenkes Mataram

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32807/jambs.v9i1.267

Transfusi darah adalah proses pemberian darah dari seseorang (donor) kepada orang lain (resipien) yang bertujuan untuk mengganti darah hilang akibat perdarahan, mengatasi shock, memelihara serta mempertahankan kesehatan resipien. Analisis pretransfusi khususnya pada uji serologi sangat diperlukan untuk menjamin darah yang ditransfusikan aman dan tidak menyebabkan reaksi transfusi. Proses pencocokan darah donor dan dengan darah pasien/resipien dilakukan melalui uji. uji silang serasi (crossmatch). Uji silang serasi dapat dilakukan melalui 2 metode yaitu yaitu menggunakan tabung dan gel card. Uji silang serasi metode tabung terdiri dari 3 fase yaitu fase 1, fase 2 dan fase 3. Pada Fase 3 merupakan tahapan yang paling berpengaruh, karena ada penambahan Anti Human Globulin (AHG). Reagen AHG berisi immunoglobulin yang berfungsi sebagai penghubung antibodi inkomplit (antibodi IgG yang menyelubungi eritrosit), sehingga akan terdeteksi melalui terbentuknya aglutinasi (gumpalan). Namun perlu diketahui bahwa ikatan antar antibodi tidak dapat bertahan lama. Ikatan antar antibodi (AHG dan antibodi inkomplit) dapat terlepas kembali (disosiasi), sehingga hasil pemeriksaan dapat menjadi false negatif. Tujuan penelitian ini adalah untuk mengetahui apakah waktu penambahan AHG mempengaruhi derajad aglutinasi pada saat uji silang serasi. Penelitian ini menggunakan metode uji silang serasi metode tabung dengan menggunakan golongan darah O rhesus positif sebagai sampel. Analisis data dilakukan melalui pengamatan derajat aglutinasi pada tiap fase uji silang serasi. Pada penelitian ini, adanya perbedaan waktu penambahan AHG mempengaruhi hasil pemeriksaan uji silang serasi. Kesimpulan dalam penelitian ini adalah efektifitas penambahan AHG pada uji silang serasi metode tabung yaitu pada menit ke 2-10 menit.

PENGARUH WAKTU SIMPAN TERHADAP PERUBAHAN pH, GLUKOSA, LDH, KALSIUM, MVP SEBAGAI INDIKATOR KUALITAS KOMPONEN DARAH THROMBOCYTE CONCENTRATE Diani Mentari; Relita Pebrina; Diah Nurpratami
Biomedika Vol 12, No 1 (2020): Biomedika Februari 2020
Publisher : Universitas Muhamadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/biomedika.v12i1.8981

ABSTRAK Thrombocyte Concentrate (TC) merupakan komponen darah yang ditranfusikan untuk pasien yang mengalami pendarahan, kelainan fungsi trombosit dan trombositopenia. Pada beberapa literatur menyebutkan bahwa sediaan TC secara in vitro dapat disimpan selama 5-7 hari. Quality Control sebelum TC ditransfusikan yaitu melalui pengamatan secara visual ada tidaknya swirling dan melihat tingkat kejernihan TC. Namun pengamatan bersifat subjektif sehingga belum terstandar. Penelitian ini bertujuan untuk melihat pengaruh waktu penyimpanan TC terhadap kualitasnya. Kualitas TC diukur melalui beberapa cara yaitu pengukuran pH, kadar glukosa, Laktat Dehidrogenase (LDH), kalsium serta profil darah yang meliputi jumlah trombosit dan Mean Platelet Volume (MPV). Analisis kadar glukosa, LDH, dan kalsium dilakukan secara kolorimetri menggunakan spektrofotomter sedangkan profil darah diukur menggunakan hematology analyzer. Hasil menunjukkan bahwa TC yang disimpan pada hari ke-9 terjadi penurunan pH sebesar 7,38%, glukosa 20,10%, LDH 42,89%dan kalsium 62,54%. Jumlah trombosit mengalami penurunan sebesar 24,41% dan MVP mengalami kenaikan 18,84%. Kesimpulan pada sampel TC yang masih terdapat swirling, namun terjadi penurunan kualitas TC yang ditandai dengan penurunan jumlah trombosit serta kenaikan nilai MVP. Selain itu semakin lama waktu penyimpanan akan menurunkan kadar pH, kadar glukosa, LDH, dan kalsium. Kata Kunci: Masa Simpan Trombosit, pH, Glukosa, Kalsium, MPV. ABSTRACT Thrombocyte Concentrate (TC) is the blood component tranfused for the patients with bleeding, abnormalities of platelet function and thrombocytopenia. In some literatures, it is mentioned that the TC preparation in vitro can be stored within 5-7 days. Quality Control before TC is transfused through a visual observation on the presence or absence of swirling and the clarity level of TC. However, this observation is subjective and makes it unstandardized. This research aims to see the effects of the shelf life of thrombocytes on its quality. The TC quality was measured through a number of ways including: pH measurement, glucose level, Lactate Dehydrogenase (LDH), calcium and profiles of blood including number of thrombocytes, and Mean Platelet Volume (MPV). The analysis on the glucose level, LDH, and calcium was conducted by colorimetrical manner using the spectrophotometer and blood profile was measured using hematology analyzer. The result showed that TC stored in day 9 experienced the decrease of pH at 7.38%, glucose at 20.10%, LDH at 42,89% and calcium at 62.54%. The number of thrombocytes experienced a decrease of 24.41% and MVP experienced an increase of 18.84%. In conclusion, the sample of TC that had swirling experienced a decrease in the TC quality as characterized with the decrease of number of thrombocytes and the increase of MVP value. In addition, the longer storage could decrease the level of pH, level of glucose, LDH, and calcium. Keywords: Platelet Storage, pH, Glucose, Calcium, MPV.

Pengaruh Perbedaan Metode Ekstraksi Metabolit Sekunder Streptomyces sp. GMR22 terhadap Toksisitas pada Sel BHK-21 Diani Mentari; Mirtani Naima; Riska Wulansari; Jaka Widada; Tri Rini Nuringtyas; Tri Wibawa; Nastiti Wijayanti
Pharmacon: Jurnal Farmasi Indonesia Vol 16, No 1 (2019)
Publisher : Universitas Muhammadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23917/pharmacon.v16i1.8032

Streptomyces sp. GMR22 is local isolate from Wanagama 1 Forest in Yogyakarta. They have the potential to be developed to produce active compounds because have PKS and NRPS genes.The active compounds from isolation are strongly influenced by various factors, one of them is extraction techniques. Effect difference of extraction technique will be affected by the quality of secondary metabolites produced.The purpose of this study was to compare the cytotoxicity effects of secondary metabolites of Streptomyces sp. GMR22 which have extracted with different stages from previous studies. The extraction technique was carried out by multilevel separatory funnel extraction methods, which was first extracted using non-polar solvent (n-hexane) and then extracted using semi-polar solvent (ethyl acetate). This research is important because in previous studies (separatory funnel only extracted using ethyl acetate) with the use of the lowest concentration in the dengue virus antiviral test (further test) caused 100% of deaths in BHK-21 cells.This study indicate that multilevel extraction result in lower CC50 value than previous studies. There are 49.160 µl/ml (n-hexane extract) and 284.56 µl/ml (ethyl acetate extract) while water extract is 464,38 µl/ml. FTIR compound analysis show that the three extracts produced have different spectrum patterns, especially in the n-hexane and ethyl acetate extract. Value of CC50 is not too high, it is expected that the secondary metabolites contained in the extracts can be used for further analysis such as antiviral testing because it is safe for normal host cells such as BHK-21 cells

PENGHAMBATAN CYTHOPHATIC EFFECT (CPE) PADA SEL BHK-21 YANG TERINFEKSI VIRUS DENGUE SEROTIPE 4 (DENV-4) DENGAN PEMBERIAN EKSTRAK N-HEKSAN STREPTOMYCES SP. GMR22 Diani Mentari; Jaka Widada; Tri Wibawa; Nastiti Wijayanti
BIOMA : JURNAL BIOLOGI MAKASSAR Vol. 5 No. 2 (2020)
Publisher : Department of Biology, Faculty of Mathematics and Natural Sciences, Hasanuddin University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20956/bioma.v5i2.10292

Negara tropis seperti Indonesia sangat berpotensi sebagai tempat berkembangnya berbagai penyakit menular berbahaya salah satunya Demam Berdarah Dengue (DBD) yang disebabkan oleh infeksi virus dengue. Variasi genetik pada serotipe virus dengue menyebabkan penyakit ini menjadi perhatian dunia medis karena menyebabkan kompleksitas respon imun yang berbeda. Vaksin Dengue (Denvaxia) diperkenalkan oleh WHO pada akhir tahun 2015, namun penggunaannya belum sepenuhnya efektif. Hal ini menyebabkan banyak peneliti berupaya untuk mencari senyawa bioaktif yang memiliki potensi untuk dikembangkan sebagai agen antiviral. Streptomyces sp. GMR22 diketahui menghasilkan senyawa bioaktif dengan spektrum yang luas. Ekstrak n-heksane Streptomyces sp. GMR22 memiliki nilai CC50 yang tidak terlalu tinggi dibandingkan dengan ekstrak Etil Asetat. Hal ini menyebabkan metabolit sekunder yang dihasilkan dapat digunakan untuk analisis lanjut seperti uji antivirus karena aman terhadap sel BHK-21 yang merupakan host virus dengue. Virus DENV-4 merupakan serotipe endemik di Asia Tenggara. Penelitian ini bertujuan untuk mengetahui aktivitas antivirus DENV-4 menggunakan ekstrak n-heksan Streptomyces sp. GMR22. Aktivitas antivirus dilakukan melalui pengamatan sel BHK-21. Hasil pengamatan morfologi sel BHK-21 terinfeksi virus DENV-4 menunjukkan bahwa pemberian ekstrak n-heksan Streptomyces sp. GMR22 mengurangi terbentuknya Cythophatic Effect (CPE). Namun penggunaan konsentrasi 40 µg/mL menyebabkan kematian pada sel BHK-21. Kata kunci: antiviral, DENV-4, metabolit sekunder, Streptomyces sp. GMR22, CPE

Human Platelet Lysate (HPL) as an Alternative Media Propagation of T47D Cells Line Diani Mentari; Relita Pebrina; Diah Nurpratami
Indonesian Journal of Cancer Chemoprevention Vol 11, No 1 (2020)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev11iss1pp36-45

Fetal bovine serum (FBS) is a gold standard as a supplement to cell and tissue culture media. This is due to a large number of Growth Factor (GF) contained in FBS. However, the use of FBS is at risk of transferring endotoxins, prions, bacteria and viruses from animals to humans, so it is risky to be used on cell therapy. Human Platelet Lysate (HPL) is a medium that can be developed as an alternative cell growth medium. The advantage of HPL is that it does not contain aggregate platelets so it does not cause the cells to clot. This condition causes HPL to be used as a substitute medium replacing FBS for cell propagation. The use of HPL for cell propagation has been widely reported. However, the use of HPL in cancer cells has not been found. Thus, this study aims to see the effectiveness of HPL as a T47D cell culture medium. The study began with donor selection with criteria for the male sex, the blood type O, the age ≤35 years. Furthermore, the Platelet Concentrate (PC) was processed into HPL then measured pH, total protein and albumin levels. The cell viability was measured using the MTT assay to determine the ability of cell proliferation when propagation using HPL. The doubling time test was carried out as in the cell proliferation test. However, the incubation was carried out for 24 h, 48 h and 72 h and the HPL concentration used was 5%. The result shows that HPL 10% and 20% ability to increase proliferation better than the FBS 10%. HPL with a 5% concentration ability to shortens the doubling time than FBS 10% (doubling time is less than 19.94 h). It this study, cell proliferation is influenced by the pH of HPL and total protein but not by the amount albumin.Keywords: Human Platelet Lysate, Proliferation, T47D cell line, total protein, albumin.

Gambaran Golongan Darah ABO-Rhesus dan Pola Sidik Jari Pada Mahasiswa Program Studi D-3 Teknologi Transfusi Darah STIKES Guna Bangsa Yogyakarta St. Raihanun; Diani Mentari; Meyta Wulandari; Relita Pebrina
Journal of Health (JoH) Vol 6 No 2 (2019): Journal of Health - July 2019
Publisher : LPPM STIKES Guna Bangsa

Human identification is the recognition of individuals based on some physical characteristics that are unique to individuals. Fingerprints are constant, individuality and form the most reliable criteria for identification. ABO-Rhesus Blood group is also one method used to identify someone, because blood type is inheritance. This research was conducted to see the description of ABO-Rhesus blood group and fingerprint patterns students D-3 Teknologi Transfusi Darah STIKES Guna Bangsa Yogyakarta. In this study using quantitative cross sectional descriptive research and blood group samples were taken using the slide method and fingerprint patterns were taken using the fingerprint method. In this study there were 78 samples, 58 females (74.36%) and 20 males (25.64%). The ABO blood group that is dominant is blood type O(35.90%), followed by blood group A(29.49%), B(28.21%), and AB(6.41%). The dominant Rhesus blood type is the positive Rhesus blood group. The percentage of fingerprint patterns in this study was loop 61.03%, whorl 37.56%, and arch 1.41%. The characteristics of the right and left hand fingerprint patterns have the same percentage of arch fingerprint patterns found on the index finger. Whorl fingerprint patterns are found on the ring finger. Loop fingerprint patterns are found on the little finger.

Utilization of Expired Platelet Concentrate for Production of Human Platelet Lysate as a Medium for T47D Cell Propagation Diani Mentari; Relita Pebrina; Diah Nurpratami
Molecular and Cellular Biomedical Sciences Vol 6, No 2 (2022)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v6i2.254

Background: Platelet concentrate (PC) has a short shelf life (5 days). Expired PC cannot be used for clinical purposes. PC is used for human platelet lysate (HPL) production, which was found to be more effective than FBS at increasing T47D cell proliferation. HPL production using expired PC has not been reported. This study aimed to investigate whether the use of HPL produced from expired PC (storage duration >5 days) can increase the proliferation of T47D cells in vitro.Materials and methods: Expired PC samples with a shelf life of 7 and 11 days were used to produce HPL via freeze/thaw method. pH, total protein content, glucose and albumin levels were measured. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to measure proliferation rate and doubling time of HPL-treated T47D cells.Results: After HPL production, the glucose level was influenced by the pH (p=0.003), and albumin level was influenced by total protein content (p=0.030). HPL stored for 7 and 11 days increased cell proliferation rate by 1.41 and 1.80 times higher than 10% FBS, respectively. HPL produced from expired PC did not cause morphological abnormality of the cells. In this study, the glucose levels affected cell proliferation (p=0.030). High glucose levels inhibited T47D cell proliferation.Conclusion: Expired PC can be used as a potential material for HPL production, since HPL produced from expired PC increases cell proliferation rate and shortens cell doubling time.Keywords: cell proliferation, human platelet lysate, platelet concentrate, thrombocyte, T47D

EFEK PEMBERIAN AIR REBUSAN DAUN SIRSAK (Annona muricata L.) TERHADAP HORMON KORTISOL PADA MENCIT YANG DIINDUKSI DEPRESI Maria Hyasinta Vanesa Putri Suhadia; Sri Luthfiyyah; Debek Bini Dita Rahayu Mayang Tika; Aurelya Putri Amanu; Diani Mentari
Borneo Journal of Biology Education (BJBE) Vol 5, No 1 (2023): APRIL
Publisher : Prodi Pendidikan Biologi Universitas Borneo Tarakan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35334/bjbe.v5i1.3633

Depresi dapat Â meningkatkan Â kadar Â kortisol Â hingga Â 20 kali. Daun sirsak (Annona muricata L.) memiliki banyak manfaat, salah satunya karena kandungan antioksidan yang terdapat di dalamnya. Penelitian ini bertujuan untuk melihat efektifitas penggunaan air rebusan daun sirsak sebagai anti depresi dengan melihat kadar kortisol pada mencit yang diinduksi depresi. Stressor dan air rebusan daun sirsak diberikan selama 7 hari. Pengukuran hormon kortisol menggunakan metode ELISA. Deteksi senyawa pada rebusan daun sirsak menggunakan metode Kromatografi Lapis Tipis (KLT) dengan pereaksi semprot FeCl3.Â Hasil analisis KLT menunjukkan bahwa pada air rebusan daun sirsak mengandung senyawa fenol. Hasil analisis hormon kortisol menunjukkan bahwa pemberian air rebusan daun sirsak mampu menghambat pembentukan kortisol pada mencit yang diinduksi stresor. Hal ini juga terlihat dari peningkatan berat badan mencit yang diberikan air rebusan daun sirsak mengalami kenaikan mencapai 15,48- 28,05% dibandingkan dengan kontrol negatif yang hanya meningkat 4,82%.

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