THE VIABILITY TEST OF SAPPAN WOOD (CAESALPINIA SAPPAN L.) ETHANOL EXTRACT IN THE H9C2 CELL LINE

Objective: In this study, the embryonic rat cardiomyocyte cell line H9C2 was used to investigate the cardiotoxicity effect of sappan wood ethanol extract (SWEE). Methods: Sappan wood was extracted in 96% ethanol and divided into dose concentrations of 2.5, 5, 10, 50, 100, and 150 μg/ml, with deferiprone used as a control. Cell viability was assessed using the PrestoBlue Cell Viability Reagent, according to manufacturer protocols. Results: Microscopic examination showed that the cell viability of H9C2 was preserved by SWEE treatments at a dose of 10 μg/ml and suggested dose concentrations of 50 μg/ml of SWEE. The percentage of viable cells was greater than 95% with a dose concentration of 10 μg/ml of SWEE, but it was significantly reduced with a dose concentration of 50 μg/ml of SWEE (p<0.05). Conclusion: The optimal dose concentration of SWEE to reach 95% cell viability was 10 μg/ml

LONG AND VERY LONG FATTY ACID FRACTIONATION IN SYSTEMIC LUPUS ERYTHEMATOSUS IN THE ACTIVE AND INACTIVE STATUS

Objective: Flare in Systemic Lupus Erythematosus (SLE) is an exacerbation of SLE clinical features that were earlier quiescent. The disease activity changes from inactive to active with an increase of several immunological profiles; the rise of immune activity induces a metabolic shift in SLE patients. The previous study aimed to investigate the long and very long fatty acid fractions (LCFA and VLCFA) in the active and inactive statuses of SLE patients and showed there were dynamic changes in fatty acid fractions in SLE patients, compared to healthy subjects. The aim of this preliminary study is to investigate LCFA and VLCFA in the active and inactive condition of SLE patients. Methods: Four serum samples of active and inactive statuses from the same SLE patients were used in this study. Serum LCFA and VLCFA fractions were analyzed by a 7890 Gas Chromatography (GC) System 5977 Mass Selective Detector (MSD). Results: All of the LCFA and VLCFA fractions were increased in the active condition, compared to SLE patients in inactive, although they were statistically not different (p>0.05). The total fatty acid fraction was 38% higher in active condition compare to inactive. The prominent increase of fatty acid fractions was alpha-linolenic acid (inactive vs. active: 23.25±17.97 vs 48.25±38.58 μmol/l), oleic acid (1300±190.4 vs 1774±866.3 μmol/l) and myristic acid (31.25±12.76 vs 59.25±40.4 μmol/l). Conclusion: The serum of LCFA and VLCFA fractions in SLE patients tend to increase in active conditions

Effects of Morinda citrifolia Leaf Extract on Glucose Absorption through Intestinal Epithelial Membrane in Wistar Rat Models

Background: Noni fruit (Morinda citrifolia) has been used as herbal medicine by Indonesian people. However, the effect of noni leaves as an antihyperglycemic agent is still unknown. This study was conducted to find out the effect of Morinda citrifolia leaves on glucose absorption through intestinal epithelial membrane in wistar rat models. Methods: The study was conducted in November 2015 at the Biochemistry and Molecular Biology Laboratory Faculty of Medicine, Universitas Padjadjaran. We used in situ perfusion and rats were divided into 2 groups. The first group used glucose solution as a control. The second group used glucose with extracts. Specimens were taken before and after perfusion to check the amount of glucose with spectrophotometer. Statistical analysis using t-test was conducted to compare the total absorbed concentration of glucose from each group. Results: From calculation we found the kinetic value (Km) of glucose absorption without extract (Km=17.24) and with extract (Km=16.67). Statistical test showed there was no significant in results (p>0.05). Conclusions: There is no effect of Morinda citrifolia leaf extract on glucose absorption. However, kinetics of glucose absorption suggested a non-competitive inhibition by Morinda citrifolia leaf extract

Old Mice Epididymal Sperm Quality After Short Term Gavage of Cogon Grass Root Ethanol Extract

Cogon grass (Imperata cylindrica) has been known as traditional herb because of its antioxidant activity. The oxidant level in the body increases linearly with age. Antioxidant works by decreasing the vulnerability of mammalian spermatozoa towards free radical attack. This study aimed to determine whether the active compound of Imperata cyclindrica root extract improved or damaged  sperm function in old mice. This was a complete randomized study performed in the Mouse Animal Laboratory Faculty of Medicine, Universitas Padjadjaran during the period of August to December 2017. This study was carried out by oral administration of three different doses i.e 115, 230, and 345 mg/kg body weight of cogon grass root ethanol extract into 24- weeks-old DDY strain mice for 14 days. Results showed no significant difference in body weight  and testicular index before and after treatment (p<0.05). Interestingly, sperm concentration decreased significantly in 345 mg/kgBW group when compared  to control (2.03x106 and 6.43x106 respectively, p<0.05). On the other hand, sperm motility and sperm with normal morphology increased following dose pattern, although not significantly, when compared to control (p<0.05). Hence, cogon grass root ethanol extract cannot maintain optimum epididymal sperm quality in old mice.  Key words: Aging, Imperata cylindrica, sperm qualit

Anemia and Eosinophilia in Traditional Goat Farmers: Early Markers of Strongyle Zoonoses (ANEMIA DAN EOSINOFILIA PADA PETERNAK KAMBING TRADISIONAL: PENANDA AWAL ZOONOSIS STRONGYLUS)

The spread of zoonoses between humans and animals can increase with the presence of specific contacts between the two. As well as intensive contact between farmer and goat. Anemia is a common hematologic change encountered in infection and zoonoses so it can be used as an early indicator for zoonoses. Nevertheless, anemia itself cannot lead to a specific etiology thus differential leukocyte count and goat fecal examination can be a good supporting test. Understanding the etiologies of the disease is critically important in preventing the decline in human’s quality of life as well as economic losses. This cross-sectional study included 30 farmers who were recruited by purposive sampling method. They agreed to do a CBC examination with 22 parameters. From the test, 37% of them showed mild anemia. There were 45% farmers with microcytic hypochromic anemia while the rest had normocytic normochromic anemia. There was an increased number in eosinophil and segmented neutrophil from anemic farmers, 18% and 9% respectively. Besides, 13% of goats physical examination indicated as below normal conditions characterized by lean bodies, lymph node swelling, and pale mucosa. Whereas 38% goat’s fecal test showed evidence of Strongyle eggs. The research suggests a possible interrelation between animals, humans, and the environment with an increased risk of zoonoses. Anemia and eosinophilia that occur can be an early marker which will be decisive not only for the success of the disease progression reduction but also the long term suppression of the economic loss

Cogongrass (<i>Imperata cylindrical</i> L.) Roots Ethanol Extract to Improve Hematological Profile in Carbon Tetrachloride-Injection Mice Model

Carbon tetrachloride (CCL4) is widely used in industry, toxic to the environment and humans, and most often used as a model of acute liver damage and liver fibrosis in experimental animals. Liver damage can deteriorate the hematological profile. The roots of cogongrass (Imperata cylindrica L.) have been used as traditional medicine due to its antioxidant activity. This study was conducted at the Faculty of Medicine, Universitas Padjadjaran, from January to March 2019. The study aimed to investigate whether the cogongrass roots ethanol extract (CGRE) can ameliorate the disturbance in the hematological profile in acute CCl4-injected mice. CGRE in dose 150 and 200 mg/kgBW was given orally to mice for four weeks before intraperitoneal injection of CCl4 1 mL/kgBW in olive oil (1:1 v/v). After 48 hours, mice were sacrificed, and the whole blood was drawn for hematological analysis. As a result, mean corpuscular volume (MCV) was reduced in CCl4-induction mice treated with CGRE in dose 150 mg/kgBW (49.25±3.06 vs 43.38±2.13 fl, p<0.05). This condition was followed by the improved hematocrit (Hct) and mean corpuscular hemoglobin concentration (MCHC). Platelet and platelet crit (Pct) levels were tended to decrease in CCl4-induction mice treated with CGRE in dose 150 mg/kgBW. In conclusion, CGRE dose 150 mg/kg BW can improve MCV, Hct, MCHC, platelet, and Pct in CCl4-injection mice. The antioxidant level in CGRE might facilitate it.   EKSTRAK ETANOL AKAR ALANG-ALANG (IMPERATA CYLINDRICAL L.) MEMPERBAIKI PROFIL HEMATOLOGI PADA MENCIT YANG DIINJEKSI CARBON TETRACHLORIDE Carbon tetrachloride (CCL4) banyak digunakan pada industri, bersifat toksik bagi lingkungan dan manusia, serta sering digunakan pada hewan coba untuk kerusakan liver akut dan fibrosis. Kerusakan liver dapat menyebabkan gangguan profil hematologi. Akar alang-alang (Imperata cylindrica L.) telah digunakan sebagai obat tradisional karena memiliki aktivitas antioksidan. Penelitian ini dilakukan di Fakultas Kedokteran Universitas Padjadjaran pada bulan Januari hingga Maret 2019. Tujuan penelitian ini adalah meneliti apakah ekstrak etanol akar alang-alang dapat memperbaiki gangguan profil hematologi pada mencit yang diinjeksi CCL4 secara akut. Ekstrak etanol akar alang-alang (EEAA) dosis 150 dan 200 mm/kgBB diberikan per oral kepada mencit selama empat minggu sebelum injeksi intraperitoneal CCl4 1 mL/kgBB yang dilarutkan dalam minyak zaitun (1:1 v/v). Setelah 48 jam, mencit dikorbankan dan diambil darahnya untuk pemeriksaan hematologi. Sebagai hasil, mean corpuscular volume (MCV) menurun pada mencit yang diinduksi CCl4 dengan perlakuan EEAA 150 mg/kgBB (49,25±3,06vs 43,38±2,13 fl, p<0,05). Keadaan ini diikuti dengan perbaikan hematokrit (Hct) dan mean corpuscular hemoglobin concentration (MCHC). Kadar platelet dan platelet crit (Pct) cenderung menurun pada mencit yang diinduksi CCl4 dengan perlakuan EEAA 150 mg/kgBB. Sebagai simpulan, EEAA dosis 150 mg/kgBB dapat memperbaiki MCV, Hct, MCHC, platelet, dan Pct pada mencit yang diinjeksi CCL4. Kemungkinan difasilitasi oleh antioksidan pada EEAA

Cogongrass (<i>Imperata cylindrica</i> L.) Ethanol Extract on Sepsis Mice Model Body Weight and Sepsis Score

Sepsis causes damage for cells, behavioral phenotype regression, and will end in most patients' death. The ethanol extract of cogongrass (Imperata cylindrica L.)  acts as an antioxidant. This study aimed to observe the effect of giving ECGR to body weight (BW) and the sepsis score of the sepsis mice model by lipopolysaccharide (LPS) induction. This study was an in vivo study with a randomized post-test controlled group design at the Animal Laboratory of Universitas Padjadjaran, 2018. We used 4 (four) groups of male mice (Mus musculus) DDY strains. Group 1 as a control, group 2: LPS 10 μL/kgBW, group 3, and 4: LPS+ECGR (90 mg/kgBW, and a dose of 115 mg/kgBW, respectively). This treatment was performed for two weeks. Every three days, we measured their body weight. After two weeks, group 2, group 3, and 4 were injected with LPS for 8 hours to induce sepsis. Next, we measured body weight and sepsis score using murine sepsis score (MSS). Then statistical analysis was performed using ANOVA and the Kruskal-Wallis test. The results showed no differences in body weight were found in the treatment groups (3 and 4) compared with control, suggesting no effect of ECGR in decreasing mice body weight. The sepsis score was more than 21 in groups treated with LPS (2, 3, and 4), suggesting LPS can induce sepsis. There was a slight decrease in scores in-group 3 and 4 compared with group 2. This study concludes that the treatment of ECGR caused no harm to body weight and slightly decreased sepsis score in the sepsis mice model.   EKSTRAK ETANOL ALANG-ALANG (IMPERATA CYLINDRICA L.) TERHADAP BOBOT BADAN DAN SKOR SEPSIS MENCIT MODEL SEPSIS Sepsis menyebabkan kerusakan sel, regresi fenotipe perilaku, dan akan berakhir kematian pada sebagian besar pasien. Ekstrak etanol akar alang-alang (Imperata cylindrica L.) (ECGR) berperan sebagai antioksidan. Penelitian ini bertujuan mengetahui pengaruh pemberian ECGR terhadap bobot badan (BB) dan skor sepsis pada mencit model sepsis yang diinduksi lipopolisakarida (LPS). Penelitian ini adalah penelitian in vivo dengan desain randomized post-test controlled group di Laboratoium Hewan Universitas Padjadjaran tahun 2018. Kami menggunakan 4 (empat) kelompok mencit jantan (Mus musculus) strain DDY. Kelompok 1 sebagai kontrol, kelompok 2 diinduksi LPS 10 μL/kgBB, kelompok 3 dan 4 diinduksi LPS+ECGR (dosis 90 mg/kgBB dan 115 mg/kgBB masing-masing). Perlakuan ini dilakukan selama 2 minggu. Setiap tiga hari dilakukan pengukuran bobot badan mencit. Setelah dua minggu, kelompok 2, kelompok 3, dan kelompok 4 diinjeksi LPS selama 8 jam untuk menginduksi sepsis. Selanjutnya, diukur bobot badan dan skor sepsis menggunakan murine sepsis score (MSS). Analisis statistik menggunakan ANOVA dan Uji Kruskal-Wallis. Hasil penelitian menunjukkan tidak terdapat perbedaan bobot badan pada kelompok perlakuan (3 dan 4) dibanding dengan kelompok kontrol yang menunjukkan ECGR tidak berpengaruh dalam menurunkan bobot badan mencit. Skor sepsis lebih dari 21 pada kelompok yang diinduksi LPS (2, 3, dan 4) menunjukkan LPS dapat menyebabkan sepsis. Terdapat sedikit penurunan skor pada kelompok 3 dan 4 dibanding dengan kelompok 2. Simpulan penelitian ini, pengobatan ECGR tidak membahayakan bobot badan dan mengakibatkan sedikit penurunan skor sepsis pada mencit model sepsis

A Critical Role of Fatty Acid Binding Protein 4 and 5 (FABP4/5) in the Systemic Response to Fasting

During prolonged fasting, fatty acid (FA) released from adipose tissue is a major energy source for peripheral tissues, including the heart, skeletal muscle and liver. We recently showed that FA binding protein 4 (FABP4) and FABP5, which are abundantly expressed in adipocytes and macrophages, are prominently expressed in capillary endothelial cells in the heart and skeletal muscle. In addition, mice deficient for both FABP4 and FABP5 (FABP4/5 DKO mice) exhibited defective uptake of FA with compensatory up-regulation of glucose consumption in these tissues during fasting. Here we showed that deletion of FABP4/5 resulted in a marked perturbation of metabolism in response to prolonged fasting, including hyperketotic hypoglycemia and hepatic steatosis. Blood glucose levels were reduced, whereas the levels of non-esterified FA (NEFA) and ketone bodies were markedly increased during fasting. In addition, the uptake of the 125I-BMIPP FA analogue in the DKO livers was markedly increased after fasting. Consistent with an increased influx of NEFA into the liver, DKO mice showed marked hepatic steatosis after a 48-hr fast. Although gluconeogenesis was observed shortly after fasting, the substrates for gluconeogenesis were reduced during prolonged fasting, resulting in insufficient gluconeogenesis and enhanced hypoglycemia. These metabolic responses to prolonged fasting in DKO mice were readily reversed by re-feeding. Taken together, these data strongly suggested that a maladaptive response to fasting in DKO mice occurred as a result of an increased influx of NEFA into the liver and pronounced hypoglycemia. Together with our previous study, the metabolic consequence found in the present study is likely to be attributed to an impairment of FA uptake in the heart and skeletal muscle. Thus, our data provided evidence that peripheral uptake of FA via capillary endothelial FABP4/5 is crucial for systemic metabolism and may establish FABP4/5 as potentially novel targets for the modulation of energy homeostasis

Perbandingan Viabilitas Oosit Pascavitrifikasi pada Dua Tingkat Konsentrasi Sukrosa yang Berbeda

Vitrifikasi merupakan suatu teknik kriopreservasi tanpa disertai pembentukan kristal es, baik intraseluler maupun ekstraseluler. Tingkat keberhasilan vitrifikasi sangat dipengaruhi oleh jenis dan konsentrasi krioprotektan yang digunakan. Sukrosa merupakan krioprotektan ekstraseluler yang mempunyai peranan dalam menjaga kestabilan membran sel pada saat proses dehidrasi. Tujuan penelitian ini adalah mengkaji efek penambahan sukrosa dalam dua level konsentrasi yang berbeda pada morfologi dan persentase hidup oosit pascavitrifikasi dengan menggunakan oosit domba yang telah dimatangkan secara in vitro sebagai model. Penelitian dilaksanakan di Laboratorium Riset dan Bioteknologi, Fakultas Peternakan, Universitas Padjadjaran periode September 2016–Desember 2016. Penelitian menggunakan rancangan acak lengkap dengan dua kelompok perlakuan, yaitu penambahan 0,5 M sukrosa atau 0,65 M sukrosa pada media vitrifikasi. Parameter yang diamati adalah persentase oosit hidup dan morfologi oosit yang mati pascavitrifikasi-pencairan kembali. Hasil yang diperoleh menunjukkan bahwa persentase oosit hidup pascavitrifikasi meningkat 20% setelah penambahan sukrosa 0,65M. Fraktur zona pelusida merupakan kerusakan morfologi yang banyak ditemukan pada oosit yang divitrifikasi dengan penambahan 0,5 M sukrosa dalam larutan vitrifikasi. Hasil penelitian ini mengindikasikan bahwa penambahan sukrosa 0,65 M sebagai krioprotektan ekstraseluler meningkatkan peluang hidup oosit setelah proses vitrifikasi.  Kata kunci: Krioprotektan, morfologi oosit, sukrosa, vitrifikasi Comparison of Oocyte Viability after Vitrification with Two Different Sucrose Concentration Level Vitrification is a cryopreservation technique without the formation of ice crystals, both intracellular and extracellular. The success of vitrification depends on the type and concentration of cryoprotectants.  Sucrose is one of the extracellular cryoprotectants that play a role in maintaining cell membrane during dehydration process. The purpose of this study was to examine the effect of the addition of two different sucrose concentration levels on the morphology and viability of oocyte after vitrification using matured sheep oocytes as a model. This study was conducted at the Research and Biotechnology Laboratory, Faculty of Animal Husbandry, Universitas Padjadjaran during September–December 2016. The oocytes were randomly assigned into groups with two different concentrations of sucrose in vitrification media: i.e. 0.5 M and 0.65 M. The parameters observed were the percentage of live oocytes and the dead oocyte morphology after vitrification-thawing.  The results showed that the percentage of live matured oocytes post-vitrification increased by 20% after the addition of 0.65M sucrose. Fracture of pellucida zone was the major finding in the post-vitrification dead oocytes  in 0.5 M sucrose group. This indicates that vitrification using 0.65M sucrose as extracellular cryoprotectant presents a higher oocyte survival after vitrification-thawing.  Key words: Cryoprotectant, oocytes morphology, sucrose, vitrificatio