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BACKGROUND Surra is an infection caused by a blood protozoan parasite, Trypanosoma evansi, and transmitted by blood-sucking insects. The parasite generally infects only animals; however, it was reported to infect an Indian cattle farmer in 2004, followed by reports of other human cases. The most severe Surra outbreak in Indonesia occurred in Sumba Island during 2010–2012, resulting in the death of more than 2,000 livestock. This study was conducted to explore the serological status of farmers who have intensive contact with their livestock against T. evansi infection in Southwest Sumba district. METHODS A total of 24 serum samples were collected from farmers living in the Surra outbreak area. All sera were tested using both card agglutination test for trypanosomiasis/T. evansi (CATT/T. evansi) and field enzyme-linked immunosorbent assay (FELISA). RESULTS Of the 24 serum samples, 4 (16.7%) samples were seropositive for the antigen T. evansi using both tests. This is the first report of human trypanosomiasis (Surra) in Indonesia. Unfortunately, the clinical manifestations of farmers with positive Surra infection were not reported because all sera samples used in this study were obtained from the Public Health Service with no reports of clinical signs from the respondents. CONCLUSIONS Farmers living in the Surra outbreak area have a high risk of being infected with T. evansi due to their potential frequent contact with Surra vectors. Therefore, T. evansi infection in humans requires attention as it might have the potential to develop as a new emerging zoonotic disease in Indonesia.

The house gecko (Hemidactylus platyurus) has evolved the ability to autotomize its tail when threatened. The lost part is then regrown via epimorphic regeneration in a process that requires high energy and oxygen levels. Oxygen demand is therefore likely to outstrip supply and this can result in relative hypoxia in the tissues of the regenerating tail. The hypoxic state is stabilized by the Hypoxia Inducible Factor-1α (HIF-1α) and HIF-2α proteins. We induced tail autotomy in 30 mal H. platyurus adults using a standard procedure and then collected samples of the regenerated tail tissue on days 1, 3, 5, 8, 10, 13, 17, 21, 25, and 30 post autotomy. For each sample, mRNA expression was analyzed by qPCR, proteins were analyzed using Western Blot tests and immunohistochemistry, and the histological structure was analyzed using Hematoxylin and Eosin staining. On day 1, HIF-1α mRNA expression increased and the tissue was dominated by leucocyte and erythrocyte cells. HIF-1α mRNA expression peaked on day 3, at which time some cells were actively proliferating, migrating, and differentiating. At the same time as HIF-1α expression decreased, HIF-2α mRNA expression increased, as did overall cellular activity. HIF-2α expression increased more gradually but was present over a longer period of time than HIF-1α. We hypothesize that HIF-1α helps to initially stimulate the tissue regeneration process while HIF-2α functionally takes over the role of HIF-1α after HIF-1α succumbs to the oxygen conditions, but we suspect that both HIF-1α and HIF-2α play a role in overcoming the tissue’s hypoxic state.

Infertility in women could be a result of an excessive production of antisperm antibody (ASA). Paternal lymphocyte immunization (PLI) could decrease the ASA levels, but the mechanism is still unclear. The aim of this study was to analyse the impact of a PLI-induced ASA decline on regulatory T-cell populations and serum interleukin-10 (IL-10) levels in women with unexplained infertility.Samples were obtained from patients who came to Sayyidah Mother and Child Hospital in Jakarta from July 2018 to April 2019 with infertility problems. The inclusion criterion for this study was unexplained infertility. Each patient was examined for ASA titres using husband's sperm auto-agglutination test (HSAaT) method, and patients with ASA titres1:128 were given PLI subcutaneously every 3 weeks. ASA titres were evaluated again 2 weeks after PLI with HSAaT. A total of 12 samples were analysed. Regulatory T-cell populations were evaluated using flow cytometry and human forkhead box P3 FoxP3 staining kit of Biotech and Device, and serum IL-10 was determined using an Abcam ELISA kit. The data were analysed using Wilcoxon and Spearman tests.PLI decreased serum ASA and percentage of regulatory T cells (p = 0.023). The decrease in ASA and subsequent decrease in regulatory T cell population was due to the strong negative correlation between regulatory T cells and IL-10 (r = -0.817, p = 0.004).The decline in ASA was associated with a decrease in regulatory T cells due to a negative correlation with IL-10levels.

Latar belakang: Otak memerlukan oksigen yang banyak selain glukosa. Hipoksia iskemik karena strok hemoragik atau Spontaneous intracerebral hemorrhage (sICH) dapat mengganggu suplai oksigen dan nutrisi ke otak berakibat produksi energi di otak akan menurun. Deplesi energi ini menyebabkan kerusakan dan kematian sel otak terjadi lebih cepat. Neuroglobin (Ngb) dan sitoglobin (Cygb) merupakan protein golongan globin yang terdapat di otak dan berperan sebagai protein pengikat oksigen di mitokondria. Tujuan: Penelitian ini bertujuan mendapatkan gambaran/profil kadar Ngb dan Cygb, di plasma, cairan serebro spinal (CSS), dan jaringan otak pasien strok hemoragik. Metode: Penelitian ini merupakan penelitian lanjutan menggunakan sampel plasma, CSS, dan jaringan otak yang diperoleh saat kraniotomi evakuasi hematoma pasien strok hemoragik sICH di rumah sakit Cipto Mangunkusumo dan rumah sakit lainnya di Jakarta. Kadar protein Ngb dan Cygb dari plasma, CSS, dan jaringan otak diukur dengan metode ELISA. Hasil: Rerata kadar Ngb otak adalah 0,058 ng/mg protein otak, sedangkan di plasma dan CSS masing-masing adalah 0,017 ng/mg protein otak dan 0,013 ng/mg protein otak atau 29,31% dan 22,41% dari rerata kadar Ngb otak. Rerata kadar Cygb otak adalah 4,943 ng/mg protein otak, di CSS adalah 1,685 ng/mg protein otak, atau 25,26% dari rerata Cygb otak, sedangkan di dalam plasma hampir tidak terdeteksi. Simpulan: Pada keadaan hipoksia oleh karena strok hemoragik sICH, protein Ngb dan Cygb dapat diukur di plasma, CSS, dan jaringan otaknya.

This study examined the hypothesis that there is an impairment of macrophageal function in spinal TB. We examined macrophageal functions in spinal TB patients. Monocytes were isolated from peripheral blood mononuclear cells (PBMCs) of five spinal TB patients and five healthy persons as control. The isolated monocytes were cultured with stimulation of macrophage colony-stimulating factor (M-CSF) for seven days for maturation. The phagocytic ability of the macrophages derived from monocytes was measured. Also, nitric oxide (NO), myeloperoxidase (MPO), beta-glucuronide, and acid phosphatase activity was investigated. We found that the monocytes collected from patient PBMCs were significantly fewer than those of the control group (2992.103 vs. 6474.103 (cells/mL)). There were also fewer macrophages that had adhered to sheep red blood cells (SRBC) (598.103 vs. 264.103 (cells/mL)). However, NO production (2346 vs. 325.17 (µmol/gram of protein)), and the MPO (570.7 vs. 17.4 (unit/mg), beta-glucuronide (0.149 vs. 0.123 (μmol/hour/100 mg of protein)), and acid phosphatase activities (1776.9 vs. 287.9 (μmol/hour/100 mg of protein)) of the macrophages in the spinal TB group were markedly higher than in the healthy group. Despite the low adhesion to foreign bodies, the intracellular processing of TB macrophages, including oxidative activity and lysosome function, was significantly high. These results suggested the impairment of macrophageal function in spinal TB. Possibly, there is a dominance of innate non-specific immunity in spinal TB infection.

Background: Lipoic acid is a substance contained in intra and extracellular that act as a coenzyme of Pyruvate Dehydrogenase, also as an antidote, chelating agent and antioxidant. Measurement of lipoic acid is needed to determine the amount of lipoic acid that performs its functions either as a coenzyme or an antioxidant. Besides, this measurement requires a special tool such as High Performance Liquid Chromatography (HPLC) and a process that is available in rural or simple laboratories. Objective: A common and easy tool such as a spectrophotometer was conducted and could expected to be a tool of lipoic acid determination in body fluid such as plasma. Methods: Measurement of lipoic acid using spectrophotometry with UV methanol and visible PdCl2 has been tested and compared to HPLC measurement that was valid and reliable in drug measurement or pharmaceutical preparations. Results: Determination of lipoic acid in plasma and leukocytes using PdCl2 produced replicable, reliability and valid result, with high accuracy, precision and was not different from lipoic acid measurement using HPLC, p=0.99. While UV methanol was different compare to HPLC p =0.0001 or was not valid. Conclusion: The measurement of lipoic acid using PdCl2 visible method can be applied to determine the levels of lipoic acid (LA) and DHLA in plasma and equal to HPLC result.

Background The tissue regeneration process requires high oxygen and energy levels. Cytoglobin (Cygb) is a member of the globin family, which has the ability to bind oxygen, plays a role in dealing with oxidative stress, and carries oxygen into the mitochondria. Energy production for tissue regeneration is associated with mitochondria—especially mitochondrial biogenesis. The peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1alpha protein helps to regulate mitochondrial biogenesis. House geckos (Hemidactylus platyurus) are reptiles that have the ability to regenerate the tissue in their tails. House geckos were selected as the animal models for this study in order to analyze the association of Cygb with oxygen supply and the association of PGC-1α with energy production for tissue regeneration. Results The growth of house gecko tails showed a slow growth at the wound healing phase, then followed by a fast growth after wound healing phase of the regeneration process. While Cygb mRNA expression reached its peak at the wound healing phase and slowly decreased until the end of the observation. PGC-1α mRNA was expressed and reached its peak earlier than Cygb. Conclusions The expressions of both the Cygb and PGC-1α genes were relatively high compared to the control group. We therefore suggest that Cygb and PGC-1α play an important role during the tissue regeneration process.

Background: Keloid is an abnormal wound healing process that extends beyond the site of injury. Keloid and tumor’s shared similarity of recurrence suggesting a shared underlying mechanism that involves stemness. Octamer-binding transcription factor-4 (Oct-4) and aldehyde dehydrogenase-1 (ALDH1) are stem cell stemness markers. This study aimed to analyze Oct-4 and ALDH1 expressions in keloid tissues.Methods: Samples were obtained from keloid tissue excisions from three keloid patients and post-circumcision preputial skin from three healthy donors (normal control) in accordance with the local ethical committee regulation. Total RNA was isolated using TriPure Isolation kit (Ameritech), and expressions of Oct4 and ALDH1 mRNA in keloid and preputial skin were determined by quantitative reverse transcription–polymerase chain reaction (qRT-PCR) using Livak method.Results: The qRT-PCR analysis revealed the expressions of Oct4 and ALDH1 in keloid and preputial skin tissues. Keloid tissues exhibited lower expression levels of Oct-4 and ALDH1 than the preputial skin. The difference was statistically insignificant.Conclusion: Keloid tissues express Oct-4 and ALDH1 as stemness markers, and the stemness characteristics of keloid might be similar to a normal skin.

The aim of this study was to investigate the influence of intermittent hypobaric hypoxia on the expression hypoxia adaptation proteins, namely hypoxia inducibla factor-1a (HIF-1a) and myoglobin (Mb). Twenty five male Sprague-Dawley rats were exposed to intermittent hypobaric hypoxia in a hypobaric chamber in Indonesian Air Force Institute of Aviation Medicine, for 49.5 minutes at various low pressure, 1 week interval for 4 times (day 1, 8, 15 and 22). HIF-1α and Mb protein were measured with ELISA. mRNA expression of Mb was measured with one step real time RT-PCR. HIF-1α protein levels increased after induction of hypobaric hypoxia and continues to decrease after induction of intermittent hypobaric hypoxia 3 times (ANOVA, p = 0.0437). mRNA expression and protein of Mb increased after induction of hypobaric hypoxia and continues to decrease after induction of intermittent hypobaric hypoxia 3 times (ANOVA, p = 0.0283; 0.0170), and both are strongly correlated (Pearson, r = 0.6307). The heart of rats adapted to intermittent hypoxia conditions by upregulation the expression of HIF-1a and myoglobin and then both return to normal level.

The maternal immune system requires tolerance for conception to occur. It is not only the balance of Th 1/Th2 that plays a role in pregnancy, but also the regulatory T cells (Tregs) that regulate the important role in pregnancy. One cause of failure in pregnancy is due to immunological factors, including antisperm antibodies (ASA). About 10-30% of infertile couples are caused by ASA. Th1 secretes Interferon γ (IFNγ). IFNγ is also an inducer indoleamin 2,3 dioksigenase (IDO). Cooperation between Tregs and IDO will induce tolerance for pregnancy.Th2 secretes most ofinterleukin (IL)10. Increased IL10 and decreased IL6 occur during pregnancy. The aim of this study was to analyse difference of Tregs, IL10, IL6, IFNγ, and IDO levels in female with high ASA and virgin.Samples with high ASA were examined ASA titres using the husband's sperm auto-agglutination test (HSAaT) method.49 samples were analysed. Tregs were evaluated using flowcytometry with the human forkhead box P3 (FoxP3) staining kit of Biotech and Device.Level of IL10, IL6, and IFNγ was determined using an Abcam ELISA kit. Level of IDO was determined using an RnD ELISA kit. The data were analysed using the Mann-whitney tests.There are differences in the Tregs population (p = 0.0000.05) but there is no difference IL10, IL6, IFNγ, and IDO levels in female with high ASA and virgin (p 0.1400.05, p 0.6800.05, p 0.2040.05, and p 0.3620.05).High ASA affects of the Tregs population but has no effect on cytokines IL10, IL6, IFNγ, and IDO.

The prevalence of obesity in adults is increasing worldwide, which is problematic since obesity is associated with degenerative diseases. Nowadays Indonesia is facing an interesting phenomenon since there are adults who have been obese since childhood and others who conversely were undernourished while young. The biological differences of these two types of obesities are not well understood. This study aims to analyse the difference in the size and number of visceral adipocytes, HIF-1α, HIF-2α and MAP1LC3A/LC3 in obese adult rat groups that were undernourished at a young age compared to groups who were normal or even already fat from childhood. We analyzed Hif-1α, Hif-2α, Lc3 mRNA by RT-qPCR; HIF-1α, HIF-2α, MAP1LC3A/LC3 protein level by ELISA. The HIF-1α and HIF-2α protein level of visceral adipocytes derived from the group of rat which were undernourished while young increased significantly compared to the group which was overnourished. The visceral adipocytes of the group which was overnourished since childhood showed an increase in Hif-2α mRNA level. The Lc3 mRNA of the rat group which were undernourished since young increased significantly compared to rat group which was obese since childhood.

BACKGROUND A number of studies on the benefits of soybean (Glycine max (L.) Merr) in the treatment of diabetes mellitus (DM) have already been conducted; however, the effects of soybean extracts on telomerase reverse transcriptase (TERT) expression in improving telomerase activity in pancreatic cells is currently unknown. This study was aimed to evaluate the effects of soybean extracts on TERT protein expression in the pancreatic β-cells of rats with DM. METHODS Sixty male Sprague-Dawley rats were randomly divided into six groups: (1) negative control (N); (2) DM rats induced by alloxan (DMA); (3) DM rats with glibenclamide (DMG; positive control); (4) DM rats with 1 mg/kgBW/day soybean extract (DM1E); (5) DM rats with 5 mg/kgBW/day soybean extract (DM5E); (6) DM rats with 25 mg/kgBW/day soybean extract (DM25E). The treatments were carried out over 28 days. The measured variables included fasting blood glucose (FBG) level, TERT protein expression, and the number of pancreatic β-cells. RESULTS All parameters were measured against the diabetes control group. The FBG levels in rats DM1E, DM5E, and DM25E were significantly reduced on the 28th day (p < 0.05). TERT protein expression and the number of pancreatic β-cells (DM25E) also showed significant improvements compared to DM rats (p < 0.05). CONCLUSIONS Soybean extracts can increase TERT protein expression in pancreatic β-cells in diabetes-induced rats.

Abstract Background Tissue regeneration is a process that need a high demand of oxygen and energy supply. Cytoglobin (Cygb) is a hexacoordinate globin superfamily that possesses strong oxygen-binding ability. Cygb also has a role in preventing cells from oxidative stress and carrying oxygen into the mitochondria. The production of energy for regeneration is associated with mitochondria, especially mitochondrial biogenesis. The peroxisome proliferator-activated receptor gamma coactivator (PGC-1α) is aprotein that plays an important role in regulating mitochondrial biogenesis. The house gecko (Hemidactylus platyurus) is a reptile with high ability for tissue regeneration in its tail. The house gecko was selected as the animal model for this research to analyse the role of Cygb which is associated with oxygen supply and PGC-1α that is related to energy production in tissue regeneration. Results The curve for the tail growth showed three different phases. Cygb mRNA was highly expressed during tissue regeneration. PGC-1α mRNA was expressed earlier than Cygb, with a lower expression but still higher than the control. Conclusions During the tail tissue regeneration process of the house gecko, the expression of Cygb and PGC-1α were dynamic and relatively higher than their expression observed in the control. Cygb and PGC-1α were suspected to have a significant role in the tissue regeneration process.

BACKGROUND Activated macrophages require increased oxygen to destroy foreign bodies, leading to an increase in the levels of reactive oxygen species (ROS). Therefore, macrophages would experience hypoxic and oxidative stress conditions at the same time. Thus, this study was aimed to evaluate the mechanism of the activated macrophages to overcoming this dual condition.METHODS The activated macrophages were harvested from the intraperitoneal cavities of 18 BALB/c mice immunized with 2% sheep red blood cells (SRBCs). The macrophage suspension was divided into four groups: control, 24, 48, and 72 hours after-immunization groups. The expressions of hypoxia-inducible factor (HIF)-1α, HIF-2α, and cytoglobin (Cygb), as markers for hypoxic condition, were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA), whereas peroxisome proliferator-activated receptor gamma coactivator (PGC-1α) protein as a marker for mitochondrial biogenesis and aerobic metabolism was measured with ELISA. The analysis of oxidative stress was conducted with the water-soluble tetrazolium salt test.RESULTS The HIF-1α mRNA expression was the highest at 24 hours, whereas the HIF-2α mRNA showed no increased expression during the observation. The Cygb mRNA decreased after 24 hours. The highest expressions of HIF-1α and HIF-2α proteins were detected at 72 hours, whereas the Cygb protein expression increased since 24 hours. The PGC-1α protein expression increased at 72 hours. The WST test showed the highest ROS level at 24 hours.CONCLUSIONS The macrophages were activated by SRBCs underwent dual hypoxia and oxidative stress condition simultaneously to overcome the foreign bodies. The macrophages overcame these stress conditions by increasing their aerobic metabolism.

Introduction Effect of calcitriol on PBMCs of healthy adults have been well studied but not much is known about its effect on the PBMCs of elderly patients with various degree of frailty syndrome and immune senescence. This study was aimed to assess the effect of in vitro calcitriol immunomodulatory effect on IL-6, IL-10 and IFN-γ in elderly patients who were fit, pre-frail and frail to see which group of patients might get the most benefit of calcitriol. Methods This study was an experimental study on the PBMCs of 24 elderly people, of which 8 subjects each were in fit, pre-frail and frail categories based on the Cardiovascular Health Study criteria. IL-6, IL-10, and IFN-γ were examined by ELISA, before and after administration of lipopolysaccharide and 100 pg/mL calcitriol into PBMC cultures in vitro. Result The mean serum vitamin D level was 26.2 (2.4) ng/ml. Vitamin D level is decreasing along with worsening of frailty status. After LPS induction, calcitriol did not reduce IL-6 and IFN-γ in all the groups. Calcitriol increased IL-10 in all groups, with the most observed change in the pre-frail group. Conclusion In vitro administration of calcitriol showed anti-inflammatory potential by increasing IL-10 mainly in pre-frail subjects.

Background: Placental hypoxia may lead to oxidative stress, which inflicts damage to capillary protein junction. The aim of this study was to evaluate altered expression of endothelial junction protein of capillaries in hypoxia condition and to observe its correlation with the incidence of intraventricular hemorrhage in premature infants.Methods: A cross-sectional study was conducted by using placental tissues of premature infants as amodel of capillary integrity (29 hypoxic and 29 non-hypoxic). Hypoxia inducible factor (HIF)-1α was measured to define placental tissue response to hypoxia; malondialdehyde (MDA) and glutathione (GSH) served as markers of oxidative stress. The expressions of junctional proteins, N-cadherin and occludin were analyzed by immunohistochemistry. Intraventricular hemorrhage (IVH) was detected by cranial ultrasound at the third day. Unpaired t test, Mann-Whitney, and Chi-square tests were used to analyze the data.Results: The HIF-1α and MDA levels were slightly, but not significantly, higher in hypoxia group {13.64±8.70 pg/mg protein and 10.31 pmol/mg tissue (ranged 1.92–93.61), respectively} compared to non- hypoxia group {10.65±5.35 pg/mg protein and 9.77 pmol/mg tissue (ranged 2.42–93.31)}. GSH levels were not different in both groups (38.14 (ranged 9.44–118.91) and 38.47(ranged 16.49–126.76) ng/mg protein, respectively. mRNA expression of N-cadherin (0.13) and occludin (0.096) were significantly lower in hypoxia comparedto non-hypoxia group (p=0,001), while protein expression of N-cadherin (3.4; 75.9; 6.9; 13.8%) and occludin (20.7; 3.4; 69.0; 3.4; 6.9%) in hypoxia group was not associated with IVH (p=0.783 and p=0.743).Conclusion: Hypoxia altered expression of endothelial junction protein in placental capillaries, but no association with intraventricular hemorrhage was observed.

Background: Cell proliferation occurs not only in normal but also in cancer cells. Most of cell proliferation inhibition can be done by inhibiting the DNA synthesis, notably by intervening the formation of purine or pyrimidine. In purine de novo synthesis, it was assumed that biotin plays a role as a coenzyme in carboxylation reaction, one of the pivotal steps in the purine de novo pathways. The aim of this study was to see the avidin potency to bind biotin and inhibit mitosis. Methods: Peripheral blood mononuclear cell (PBMC) was cultured in RPMI-1640 medium and stimulated by phytohemagglutinin (PHA) in the presence or absence of interleukin-2 (IL-2), with or without avidin. The effect of avidin addition was observed at 24, 48, and 72 hours for cell proliferation, viability, and cell cycle. Statistical analysis was done by one-way ANOVA. Results: Avidin inhibited cell proliferation and viability in culture under stimulation by PHA with and without IL-2. Cell cycle analysis showed that avidin arrested the progression of PBMC after 72 hours of culture. Most cells were found in G0/G1 phase. Conclusion: Inhibition of biotin utilization by avidin binding can halt cell proliferation.

Background: Urea biosynthesis is a very important process in the liver which needs ATP, CO2 and functional mitochondria or aerobic condition. Liver can adapt to hypoxic condition, generally and locally. This study aimed to analyze the effect of chronic hypoxia on liver urea biosynthesis as indicated by the level and specific activity of mRNA of carbamoyl phosphate synthetase 1 (CPS1), a key enzyme in urea biosynthesis in hypoxic rats.Methods: 20 male Sprague-Dawley rats were placed in hypoxic chamber supplied by a mixture of 10% O2 and 90% N2. Five rats were sacrificed at 1, 3, 5, and 7 days after exposure. Liver homogenates were analyzed for HIF-1 (hypoxia inducible factor-1) by ELISA, CPS1 mRNA by real time RT-PCR and CPS1 enzymatic specific activities by Pierson method. Data were analyzed by ANOVA test and Pearson correlation.Results: The HIF-1 in liver increased significantly, as well as CPS1 mRNA and CPS1 enzymatic activities (p

The background of this research is the lack of movement or sports activities among students. The purpose of this study was to determine how much the level of physical fitness of the third semester students of the Physical Education Study Program FKIP Majalengka University 2019/2020. The method used in this research is descriptive method. The population in this study were 63 students from semester 3 A-C Physical Education Study Program, Majalengka University. The research sample was determined by incidental sampling. The research sample was 50 students. The instrument in this study was the bleep-test. To analyze the data obtained using relative frequency distribution analysis. Based on the results of the study, it shows that there is a difference between students who often do physical activity (sports) and those who rarely do physical activity (sports), the level of physical fitness is better for students who often exercise. Thus, physical activity greatly affects the physical fitness of the students themselves. Therefore, to get good fitness, students are required to be more diligent in doing physical activity (sports), so that they can move smoothly and not get sick easily. Key word: Physical fitness; College student ABSTRAK. Latar belakang penetitian ini adalah kurangnya aktivitas gerak atau olahraga dikalangan mahasiswa. Tujuan dari penelitian ini adalah untuk mengetahui seberapa besar tingkat kebugaran jasmani mahasiswa semester III prodi penjas FKIP Universitas Majalengka 2019/2020. Metode yang digunakan dalam penelitian ini adalah metode deskriptif. Populasi dalam penelitian ini adalah 63 mahasiswa dari semester 3 A - C Prodi Pendidikan Jasmani Universitas Majalengka. Sampel penelitian ini ditentukan dengan Sampling insidental . Sampel penelitian sebanyak 50 mahasiswa. Instrumen dalam penelitian ini adalah bleep - t est . Untuk menganalisis data yang diperoleh menggunakan analisis distribusi frekuensi relatif. Berdasarkan pada hasil penelitian menunjukkan bahwa adanya perbedaan antara mahasiswa yang sering melakukan aktivitas fisik (olahraga) dengan yang jarang melakukan aktivitas fisik (olahraga), level kebugaran jasmaninya pun lebih baik mahasiswa yang sering berolahraga. Dengan demikian bahwa aktivitas fisik sangat mempengaruhi kebugaran jasmani mahasiswa itu sendiri. Maka dari itu juga mendapatkan kebugaran yang baik mahasiswa dituntut untuk lebih rajin melakukan aktivitas fisik (olahraga), agar bisa beraktivitas dengan lancar dan tidak mudah sakit. Kata Kunci : Kebugaran Jasmani; Mahasiswa.

Induction of sublethal hypoxia provides neuroprotective effect pathological processes such as myocardial infarc and stroke. This study aimed to know the effect of intermittent hypobaric hypoxia (IHH) induction on cognitive function and glutamate receptor in the rat brain. The study was conducted at dr. Saryanto Institute for Aviation and Aerospace Health (LAKESPRA) from 2015-2016. 25 sprague dawley rats, divided into 4 IHH treatment groups and 1 control group. Group 1 was induced with 1x IHH exposure, group 2 was induced with 2x IHH exposure, group 3 was induced with 3x IHH exposure and group 4 was induced with 4x IHH exposure (days 1, 8, 15 and 22). After induction, cognitive function tests are performed with Y Maze and immunohistochemistry is performed to evaluate the glutamate receptor N-methyl-D-aspartic acid (NMDA). There were no significant differences between the four treatment groups compared with the control group (p>0.05) on cognitif function, but significant difference in NMDA receptor. In conclution, HHI induction was not cause change cognitive fuction but increased the response of neuroplasticity of sprague-dawley rats brain tissue which assessed from increasing NMDA receptor.&nbsp

Allergy diseases tend to increase with variety of clinical manifestations. To date, pharmacotherapy do not treat the underlying disease. Many researches concerned to study new approach of allergy therapy which need allergy mouse model for evaluation. This study was aimed to compare the effectiveness of two treatment methods by using ovalbumin for allergy sensitization. This study was conducted in PT.Bimana Indomedical animal facility Bogor on June until September 2016. Six mice were divided into 2 groups. First group was sensitized by ovalbumin 100ug subcutaneous (sc) day-0, 50 ug intra peritoneal (ip) day-14 and 100 ug intranasal (in) day-21. Second group was sensitized by ovalbumin 10 ug ip day-0 and day-7, 10 ug in day-21, 22, and 23. Body weight, hematology parameters (red blood cells, white blood cells, hemoglobin and hematocrit) and total IgE concentration of day-0 (pre-treatment) and day 24 (post-treatment) were measured. The results showed that there was an increase of mice body weight of both treatment group, while hematology changes of pre and post treatment were not significantly different (paired t-test). Whereas the increase in total IgE pre and post treatment was significantly different between the two groups (paired t-test). In conclusion, the second allergy sensitization method is better than the first allergy sensitization method.&nbsp

Background: Folate is an important substance used for purine and pyrimidine nucleotide synthesis. One measurement of folate that already establishes is using ELISA (Enzyme-linked immunosorbent assay) method. Folate binding protein is a protein that can bind folate, therefore it considered can be used as a tool that can replace antibody dependent ELISA method. Objectives: The aim of this research was to create a method for folate measurement in serum called Enzyme-labeled protein ligand binding assay (ELPLBA) by replacing antibody as used in ELISA method with folate binding protein (FBP) that purified from the whey of milk. Methods: The method is tested using 20 serum samples and compared to ELISA. Folate binding protein was purified from bovine’s milk using ammonium sulfate up to 90% saturated, DEAE-cellulose anion exchange chromatography and affinity chromatography. SDS-PAGE and western blot were used to establish the protein band of FBP that has molecular weight of ~25-35 kDa. ELPLBA was arranged with stationary phase using aminohexyl-agarose, and folic acid linked on it using carbodiimide. Results: The result show there was no significant difference of folate concentration between ELPLBA (14.804 ± 2.795) and ELISA method (13.859 ± 3.638), p = 0.363. Conclusion: ELPLBA method show similarity for determination of folate in serum which was the same as standard folate measurement (ELISA).

Background: Lymphocytes activated by immunization must increase their metabolism to meet the energy requirements for mitosis, differentiation, and protein synthesis, which may subject the cell to conditions of relative hypoxia and oxidative stress. This study was conducted to investigate the increase in the levels of transcription factors involved in both conditions.Methods: Male Balb/c mice were divided into the following four groups, each consisting of six animals: the control and three experimental groups. The experimental groups were immunized by injection of 0.2 ml of 2% sheep red blood cells (SRBC) suspended in phosphate-buffered saline (PBS). Lymphocytes were harvested from the spleens of each group at time intervals of 24-, 48-, and 72-h post-immunization. The buffy coat from splenocytes was separated using Ficoll Histopaque as the medium. The lymphocytes were separated from adherent cells by incubating the purified splenocytes in microtubes for 2-h. Cells were lysed by three freeze–thaw cycles (−80°C and 37°C) and used to analyze the levels of HIF-1α and HIF-2α (mRNA and protein), Nrf2 (protein), and glutathione peroxidase (GPx) activity.Results: The treatment caused an increase in GPx activity and HIF-1α protein concentration 24-h post-immunization, whereas the HIF-1α mRNA levels remained static. Elevated Nrf2 protein levels were detected within 48-h after treatment. Meanwhile, the HIF-2α mRNA and protein levels increased within72-h after immunization.Conclusion: Immunization with SRBC suspension induced relative hypoxia, elevated reactive oxygen species (ROS), and oxidative stress in the lymphocytes as indicated by the increase in both HIF-1α and HIF-2α protein and mRNA levels, GPx activity, and Nrf2 protein levels.

Background: High altitude can cause hypobaric hypoxia (HH), resulted from the lower barometric pressure and hence partial pressure of oxygen. Hypoxia can lead to a lot of deleterious molecular and cellular changes, such as generation of free radicals or reactive oxygen species (ROS). Increasing of ROS can cause oxidative stress if the antioxidant enzyme does not increase simultaneously. Oxidative damage in brain has toxic effect on cognitive functions. Objective: In this study, we investigate effect of acute intermittent HH on oxidative stress and antioxidant enzyme activity in rat brain. Method: Wistar rats divided into 5 groups, consisting control group and four experimental groups which treated to HH. Rats were exposed to simulated HH equivalent to 35.000 feet in hypobaric chamber for 1 minute, repeated once a week. Results: Level of malondialdehyde and carbonyl in rat brain under acute HH increased at HH exposure (group I) compare to control group. These levels decreased afterward at intermittent HH exposure (group II-IV). Specific activity of superoxide dismutase (SOD) shows increasing level at intermittent HH exposure, especially group IV was increasing of SOD level significantly. The increasing pattern of specific activity of catalase was inversely from SOD pattern, but it still has higher activity in intermittent HH compare to control group. Conclusion: Brain tissue seems to be able to perform an adequate adaptive response to hypobaric hypoxia after the training, shown by its significantly decreased MDA and carbonyl level and also increased specific activity of SOD and catalase.

Background: Hypoxia can cause acidosis. Kidney plays an essential role in maintaining acid-base balance, which involves the activities of carbonic anhydrase (CA) and glutaminase (GLS). This study is aimed to determine the expression and activities of the CA9 and GLS1 enzymes in relation to hypoxia inducible factor-1α (HIF-1α), a transcription factor protein which is a marker of hypoxia.Methods: This study was an in vivo experimental study with coupled paralel design. used 25 male Sprague-Dawley rats weighing 150-200 g. Rats were divided into 5 groups: the control group (normoxic condition) and 4 treatment groups. The latter were kept in a hypoxic chamber (10% O2: 90% N2) for 1, 3, 5 and 7 days. All rats were euthanized after treatment, kidneys excised, tissues homogenized and investigated for gene expression of CA9, GLS1 and HIF-1α. On protein level, total enzymatic activities of CA and GLS and protein of HIF-1α were also investigated. Data were analyzed statistically using ANOVA for significance, and as its alternative, used Mann-Whitney and Kruskal-Wallis test.Results: Results showed that HIF-1α mRNA increased during hypoxia, but not HIF-1α protein. It seemed that acidosis occurs in kidney tissue, indicated by increased CA9 and GLS1 mRNA expression and specific activity of total CA and GLS1. Expression of CA9 and GLS1 mRNA both showed strong positive correlation with HIF-1α mRNA, but not with HIF-1α protein.Conclusion: It is suggested that during chronic systemic hypoxia, gene expression of CA9 and GLS1 and their enzyme activities were increased as a response to acidosis and related with the expression of HIF-1α mRNA.

The purpose of this study was to get optimum medium composition and agitation to trypsin- like protease production by Lactobacillus plantarum FNCC 0270. The medium composition and agitation for enzyme production was optimized using Central Composite Design and Response Surface Method with Design Expert software version 7.1.5 Fermentation was carried out in erlenmeyer flasks at initial pH 8, 37 ° C , using an incubator shaker at 87.5 rpm. The best results showed an enzyme activity of 1.0 mU / mL , a protein level of 0.557 mg/mL, and desirability value of 0.740. Numerical optimization was performed to approach the ideal state of the fermentation or the desirability value of 1 . The medium composition containing of 3.64% baker's yeast, 1.21% glucose, and 0.13% skim milk was used for the fermentation. The enzyme activity of 1.51 mU/mL and protein level of 0.205 mg/mL can be achieved. After numerical optimization, the fermentation process was verified in erlenmeyer flasks with incubator shaking at 77 rpm, initial pH 8, 37 ° C, and 15 h fermentation. The verification results showed that the enzyme activity of 1.273 ± 0.227 mU/mL and protein level of 0.248 ± 0.012 mg/mL.

Plasmodium falciparum is an apicomplexan parasite that causes the most severe malaria in humans. Due to a lack of effective vaccines and emerging of drug resistance parasites, development of drugs with novel mechanisms of action and few side effects are imperative. To this end, ideal drug targets are those essential to parasite viability as well as absent in their mammalian hosts. The mitochondrial electron transport chain (ETC) of P. falciparum is one source of such potential targets because enzymes, such as L-malate:quinone oxidoreductase (PfMQO), in this pathway are absent humans. PfMQO catalyzes the oxidation of L-malate to oxaloacetate and the simultaneous reduction of ubiquinone to ubiquinol. It is a membrane protein, involved in three pathways (ETC, the tricarboxylic acid cycle and the fumarate cycle) and has been shown to be essential for parasite survival, at least, in the intra-erythrocytic asexual stage. These findings indicate that PfMQO would be a valuable drug target for development of antimalarial with novel mechanism of action. Up to this point in time, difficulty in producing active recombinant mitochondrial MQO has hampered biochemical characterization and targeted drug discovery with MQO. Here we report for the first time recombinant PfMQO overexpressed in bacterial membrane and the first biochemical study. Furthermore, about 113 compounds, consisting of ubiquinone binding site inhibitors and antiparasitic agents, were screened resulting in the discovery of ferulenol as a potent PfMQO inhibitor. Finally, ferulenol was shown to inhibit parasite growth and showed strong synergism in combination with atovaquone, a well-described anti-malarial and bc1 complex inhibitor.

Background: Glioblastoma multiforme (GBM) is a primary malignant brain tumor which has poor prognosis. High incidence of oxidative stress-based therapy resistance could be related to the high antioxidant status of GBM cells. Our previous study has reported that manganese superoxide dismutase (MnSOD) antioxidant expression was significantly higher in high grade glioma than in low grade. The aim of this study was to analyze the impact of MnSOD suppression toward GBM cell survival.Methods: This study is an experimental study using human glioblastoma multiforme T98G cell line. Suppression of MnSOD expression was performed using in vitro transfection MnSOD-siRNA. The MnSOD expression was analyzed by measuring the mRNA using real time RT-PCR, protein using ELISA technique, and specific activity of enzyme using inhibition of xantine oxidase. Concentration of reactive oxygen species (ROS) intracellular was determined by measuring superoxide radical and hydrogen peroxide. Cell survival was analyzed by measuring viability, proliferation, and cell apoptosis.Results: In vitro transfection of MnSOD-siRNA suppressed the mRNA, protein, and specific activity of MnSOD. This treatment significantly increased the concentration of superoxide radical; however, it did not influence the concentration of hydrogen peroxide. Moreover, viability MnSOD-suppressing cell significantly decreased, accompanied by increase of cell apoptosis without affecting cell proliferation.Conclusion: The suppression of MnSOD expression leads to decrease glioblastoma multiforme cell survival, which was associated to the increase of cell apoptotic.

Introduction: Spleen is the largest lymphatic organ in the body that is responsible for initiating an immune response to antigens, consistof macrophages and lymphocytes. Macrophages are cells that engulf foreign bodies by migration, phagocytosis, O2 burst, lysosomes fusion, synthesis and secretion of various cytokines which would require enormous amounts of energy. Therefore, macrophages needs of O2to increase resulting in hypoxia relative. To overcome this situation, the cell is equipped with a special mechanism which is under the control of HIF. They are highly dependent to O2existence to produce energy. This process is important for biochemical adaptation in homeostasis. Macrophages expressing Cygb for supplying O2 and PGC-1 for mitochondrial biogenesis in producing energy. This study aims to determine whether the spleen macrophages in mice being immunized intraperitoneally with SRBC increase the expression of HIF, Cygb and PGC-1?. Method: 24 male BALB/c mice, aged 2 months were immunized by injecting 0.2 ml SRBC 2%. Macrophages were taken from spleen of mice. Expression of mRNA and protein levels of HIF-1?, HIF-2?, Cygb, PGC-1? in macrophages were measured by real time RT-PCR and ELISA, respectively. Level of O2burst were measured with WST Salts. Observations were made at 24, 48, and 72 hours post-immunization. Results: The ability to oxidize antigen levels (O2burst) found higher at 24 hours after immunization. The levels of protein and mRNA expression of HIF-1? and HIF-2? showed the highest increase at 24 and 48 hours after immunization. The levels of Cygb protein increased at 48 hours group after immunization while mRNA expression increased at 24 hours after immunization. PGC-1? protein levels decreased in 24 hours after immunization and then increased gradually. Conclusion: This study shows the expression pattern of several proteins in spleen macrophages of mice that was immunized

Background: Tumor tissue usually became hypoxic due to disruption of oxygen supply. Adaptation response to hypoxia is mediated by transcription factor, hypoxia- inducible factor-1α (HIF-1α). HIF-1α signaling is known to increase the expression of pro-apoptotic protein cytochrome-c, and anti- apoptotic survivin. In this study we wanted to analyze the role of HIF-1α on breast cancer cells survival through pro-apoptosis cytohrome-c and anti-apoptosis survivin regulation. Methods: Breast cancer cell lines T47D were induced by CoCl 2 then harvested to analyze the expression of HIF-1α, protein cytochrome-c, mRNA survivin and cell viabilities Results: HIF-1α induction by CoCl2 causes the increase of protein and mRNA of HIF-1α, cytochrome-c protein, and survivin mRNA, but does not cause the changes in cell viability. Conclusion: HIF-1α induction have no effects on breast cancer cell line T47D viabilities due to the balance regulation between pro-apoptosis expression cytochrome-c and anti-apoptosis survivin.

Background: Liver is sensitive against hypoxia and hypoxia will stabilize HIF-1α. At the same time, hypoxia will produce reactive oxygen species (ROS) which can be scavenged by Cygb. The purpose of our study is to know, if normobaric hypoxia can induce Cygb expression and its association with HIF-1α stabilization.Methods: This is an experimental study using 28 male Sprague-Dawley rats, 150-200 g weight. Rats are divided into 7 groups: control group and treatment groups that are kept in hypoxic chamber (10% O2: 90% N2) for 6 hours, 1, 2, 3, 7 and 14 days. All rats are euthanized after treatment and liver tissue are isolated, homogenized and analyzed for oxidative stress parameter, expression of Cygb and HIF-1α.Results: Expression of Cygb mRNA and protein was increased on the day-1 after treatment and reach the maximum expression on the day-2 of hypoxia treatment. But, the expression was decreased after the day-3 and slightly increased at the day-14 of hypoxia. The correlation between expression of Cygb and oxidative stress parameter was strongly correlated. Cygb mRNA, as well as protein, showed the same kinetic as the HIF-1, all increased about day-1 and day-2.Conclusion: Systemic chronic hypoxia and/or oxidative stress up-regulated HIF-1α mRNA which is correlated with the Cygb mRNA and protein expression. Cygb mRNA as well as Cygb protein showed the same kinetic as the HIF-1, all increased about day-1 and day-2 suggesting that Cygb could be under the regulation of HIF-1, but could be controlled also by other factor than HIF-1.

Background: Tumor tissue usually became hypoxic due to disruption of oxygen supply. Adaptation response to hypoxia is mediated by transcription factor, hypoxia- inducible factor-1α (HIF-1α). HIF-1α signaling is known to increase the expression of pro-apoptotic protein cytochrome-c, and anti- apoptotic survivin. In this study we wanted to analyze the role of HIF-1α on breast cancer cells survival through pro-apoptosis cytohrome-c and anti-apoptosis survivin regulation.Methods: Breast cancer cell lines T47D were induced by CoCl2 then harvested to analyze the expression of HIF-1α, protein cytochrome-c, mRNA survivin and cell viabilities.Results: HIF-1α induction by CoCl2 causes the increase of protein and mRNA of HIF-1α, cytochrome-c protein, and survivin mRNA, but does not cause the changes in cell viability.Conclusion: HIF-1α induction have no effects on breast cancer cell line T47D viabilities due to the balance regulation between pro-apoptosis expression cytochrome-c and anti-apoptosis survivin.

Background: Mesenchymal stem cell (MSC) has been reported to have immunomodulator capacity against autoimmune diseases and to prevent allogenic tissue rejection. Many studies revealed that MSC’s inhibit T cell proliferation and induce immunosuppressive condition through the production of prostaglandins, and interleukin-10. In addition, MSC was reported to reduce circulating autoantibody in autoimmune patients following MSC transfusion. So far, there has been no report stating the presence of Fc receptors (receptors for immunoglobulin) on MSCs. The aim of this study was to reveal the expression of FcγRs in lipoaspirate-derived MSCs by measuring transcription of FcγR mRNA and whether the expression can be modulated.Methods: Lipoaspirate-derived MSCs were cultured in suitable medium and confirmed to be MSCs according to the criteria published by International Society for Cellular Therapy. Total mRNA of MSCs was isolated, and detection of human FcγRI, FcγRIIA and FcγRIIB mRNA was performed. Further, modulation of the expression was tested using heat aggregated gamma globulin (HAGG) and interferon (IFN)γ.Results: FcγRs mRNA was detected in the first passage of MSCs. However, the expression was no longer present after more than 4 passages. Further, increased level of FcγRI and FcγRIIA mRNA expression was detected with the addition of IFNγ in the culture. This preliminary finding opens a new insight for the understanding of interaction between MSCs and immunoglobulin G through FcγRs.Conclusion: Lipoaspirate-derived MSCs express FcγRs, and the expression is modulated by IFNγ.

Background: Regular physical activity is highly recommended in preventive, curative, and rehabilitative programs in order to promote health, especially cardiovascular health. However, physical activity can also cause sudden death. In athletes, sudden death may occur during sport competitions, with myocardial infarction as the most common etiology. It is suspected that continuous training without any rest-day play a role in cardiac muscle damage and sudden death during competition. Our study was aimed to learn about cardiac muscle adaptation on continuous aerobic and anaerobic physical activity without any rest-day. Methods: The specimens in our study were cardiac muscle tissue obtained from rats that had performed aerobic and anaerobic physical activity on treadmill for 1, 3, 7, and 10 days without any rest-day. Blood gas analysis and hematological assessment were used as parameters of systemic adaptation to hypoxia during physical activity. Moreover, histopathology of cardiac muscle tissue was performed as parameter for cardiac muscle damage.Results: The results showed that aerobic and anaerobic physical activity caused a systemic hypoxic condition and triggered adaptation responses. Cardiac muscle damage occurred on the 10th day in both treatment groups, with more severe damage observed in the group with anaerobic physical activity. The tissue protein level in the anaerobic group increased progressively on the 10th day.Conclusion: Physical activity may result in hypoxia and systemic adaptation. Aerobic and anaerobic physical activities performed for 10 days without any rest-day may cause cardiac muscle damage. (Med J Indones. 2013;22:209-14. doi: 10.13181/mji.v22i4.601)Keywords: Cardiac muscle, cardiac muscle damage, histopathology, physical activity

Oxidative stress is considered an important factor in the development of endometriosis, including its malignant transformation. Previous studies have found that AT-rich interactive domain 1A (ARID1A), a tumor suppressor gene, is frequently mutated and inactivated in endometriosis-associated ovarian cancer (EAOC), and such a change in this gene is considered an early event in malignant transformation. We observed oxidative stress status by measuring the activity of the antioxidant enzyme manganese superoxide dismutase (MnSOD), malondialdehyde (MDA), and ARID1A gene expression in tissue samples from patients with endometriosis, EAOC, or non-endometriosis-associated ovarian cancer (non-EAOC). We also induced oxidative stress in the cultured cells from patients with primary endometriosis by adding H2O2 and tested for any alteration of ARID1A gene expression based on different H2O2 concentrations. The results showed that MnSOD activity in endometriosis and EAOC was lower than in non-EAOC, but MDA levels were higher. This study also showed that oxidative stress reduced ARID1A gene expression.

There has been no study conducted about the effect of autosuggestion on quality of life for geriatric patients. Our aim was to evaluate the efficacy of autosuggestion for geriatric patients' quality of life and its impact on psycho-neuro-endocrine-immune pathway.Sixty geriatric patients aged ≥60 years in a ward were randomly assigned to either receive autosuggestion or not. Autosuggestion was recorded in a tape to be heard daily for 30 days. Both groups received the standard medical therapy. Primary outcome was quality of life by COOP chart. Secondary outcomes were serum cortisol level, interleukin-2, interleukin-6, interferon-γ, and N-acetylaspartate/creatine ratio in limbic/paralimbic system by magnetic resonance spectroscopy. The study was single blinded due to the nature of the intervention studied.Out of 60 subjects, 51 finished the study. The autosuggestion group reported better scores than the control one for quality of life, COOP chart 1.95 vs. 2.22 (95% CI, p = 0.02). There were increments of serum cortisol (p = 0.03) and interleukin-6 in the autosuggestion group (p = 0.04). Interleukin-2, interferon-γ, and N-acetylaspartate/creatine ratio in prefrontal cortex showed a tendency to increase in the autosuggestion groups.Autosuggestion is associated with improvement of geriatrics' quality of life, serum cortisol level, and adaptive immunity. There is a better trend for neuroplasticity in prefrontal cortex in the autosuggestion group.

This research addresses the implementation of encryption and digital signature technique for electronic health record to prevent cybercrime problem such as robbery, modification and unauthorized access. In this research, RSA 2048-bit algorithm, AES 256-bit and SHA 256 will be implemented in Java programming. Secure Electronic Health Record Information (SEHR) application design is intended to combine given services, such as confidentiality, integrity, authentication, and non-repudiation. Cryptography is used to ensure the file records and electronic documents for detailed information on the medical past, present and future forecasts that have been given only for the patients. The document will be encrypted using an encryption algorithm based on NIST Standard. In the application, there are two schemes, namely the protection and verification scheme. This research uses black-box testing and white-box testing to test the software input, output, and code without testing the process and design that occurs in the system. We demonstrated the implementation of cryptography in Secure Electronic Health Record Information (SEHR). The implementation of encryption and digital signature in this research can prevent archive thievery which is shown on implementation and is proven on the test.

Background: Exercise increases the need for oxygen to generate ATP through oxidative phosphorylation. If the high energy demand during exercise is not balanced by sufficient oxygen supply, hypoxia occurs in skeletal muscle tissue leading to upregulation of hypoxia inducible factor-1α (HIF-1α). The activity of HIF-1α increases the expression of various genes in order to reduce the metabolic dependence on oxygen and to increase oxygen supply to the tissue, e.g., VEGF which plays a role in angiogenesis. In myocardium, it is unclear whether exercise leads to hypoxia and whether HIF-1α and VEGF play a role in the mechanism of hypoxic adaptation. This study aimed to investigate the correlation of HIF-1α and VEGF in heart muscle tissue of rats during aerobic and anaerobic exercise.Methods: A rat treadmill was used with a specific exercise program for 1, 3, 7 and 10 days. The concentrations of HIF-1α and VEGF were measured the myocardium.Results: Both, HIF-1α protein and VEGF were increased (p < 0.05) in the groups with aerobic and anaerobic exercise. Concentrations of HIF-1α were highest on the first day of activity, being higher in the anaerobic than in the aerobic group (156.8 ± 33.1 vs. 116.03 ± 5.66). Likewise, the highest concentration of VEGF in the group with anaerobic exercise occurred on the first day (36.37 ± 2:35), while in the aerobic group, VEGF concentration was highest on day 3 (40.66 ± 1.73). The correlation between the myocardial tissue consentrations of HIF-1α and VEGF is moderate (r = 0.59) in the aerobic group and strong in the anaerobic group (r = 0.69).Conclusion: Aerobic and anaerobic exercise increase HIF-1α and VEGF concentrations in rat myocardium in specific patterns. The anaerobic condition triggers vascularization stronger and obviously earlier than aerobic exercise. (Med J Indones. 2012;21:133-40)Keywords: Exercise, HIF-1α, myocardium, VEGF

Background: Amaranthus spinosus and Andrographis paniculata are traditionally used as antimalarial herbs, but the combination of both has not yet been tested. The aim of this study was to determine the schizonticidal anti-malaria effect of a combination in Plasmodium berghei-infected mice.Methods: Male mice (Balb/c strain) weighing 28-30 g, 7-8 weeks old, were randomly devided into 5 groups of 4 animals each. Group A: controls (nil) and 4 treatment groups (B, C, D, and E). Group B: Amarathus 10 mg/kgBW, group C: Andrographis 2 mg/kgBW, group D: combination of Amaranthus + Andrographis 10 mg + 2 mg/kgBW. All treatment with plant extracts was administered orally, once per day for 7 days. Group E was given chloroquine 10 mg/kgBW, once a day orally, for 3 days.Results: The body weigh increased only in group D, hemoglobin concentration increased significantly vs controls (p < 0.05) in treatment groups C, D, and E, and blood schizonticidal activity was seen in all treatment groups, highest at almost 90% in groups D and E. Survival rate was 100% in all groups.Conclusion: The combination of Amaranthus and Andrographis (10 mg + 2 mg/kgBW) exerts the same blood schizonticidal activity as chloroquine 10 mg/kgBW. (Med J Indones. 2012;21:66-70)Keywords: Amaranthus spinosus, Andrographis paniculata, Balb/c mice, Plasmodium berghei, schizonticidal effect

Chloroquine (CQ)-resistant Plasmodium vivax was first documented in 1989 and threatens much of eastern Indonesia, with > 50% of therapeutic failure rates. We screened 2236 subjects for malaria infection through active case detection and identified 232 infected cases with 100 subjects carried P. vivax mono infection. We prospectively evaluated therapeutic responses to CQ in 73 subjects infected by P. vivax in northeastern Papua, Indonesia. We phenotyped these infections as susceptible or resistant to CQ using a 28-day in vivo test format. Eighteen subjects (25%) had persistent or recurrent parasitemia during the test and were provisionally classified as resistant. Among the remainder, 46 (63%) subjects had no persistent or recurrent parasitemia and were classified as having infections sensitive to CQ, 4 were lost to follow up, and 5 dropped out. Among the 18 provisionally resistant cases, 1 subject (6%) had persistent parasitemia at Day 3 and was considered as a direct treatment failure, 2 subjects (11%) had recurrent parasitemia by Day 7 and were considered early treatment failures, and 7 (39%) and 8 (44%) had recurrent parasitemia by Days 14 and 28, respectively. Analysis of blood for CQ+N-desethylchloroquine (DCQ) levels on day of recurrence from 15 of the 18 with treatment failures showed 11 subjects having CQ+DCQ blood levels ⩾ 100 ng/ml and 2 with CQ+DCQ blood levels

Background: Intermittent hypobaric hypoxia is suggested to possess a protective effect toward the hypoxic condition. The aim of this study is to analyze the expression of cytoglobin (Cygb), neuroglobin (Ngb) and the specific activity of acetylcholinesterase (AChE) in brain tissue as adaptive responses to intermittent hypobaric hypoxia. Methods: Twenty-five adult Sprague-Dawley male rats were divided into 5 groups: 1) The control group (normoxia); 2) group exposed to acute hypobaric hypoxia (AHH); 3) group exposed to hypobaric hypoxia (HH) on day-1 and re-exposed on day-8 (intermittent hypobaric hypoxia once or IHH1x); 4) group that is exposed to HH on day-1, re-exposed to HH on day-8 and day-15 (intermittent hypobaric hypoxia two times or IHH2x); 5) group exposed to HH on day-1, re-exposed to HH on day-8, day-15 and day-22 (intermittent hypobaric hypoxia 3x or IHH3x). Homogenized brain tissue was then measured and analyzed for Cygb and Ngb protein expression, and also AChE specific activity. Results: Cytoglobin and Ngb were decreased in the acute induction and increased significantly along with the increasing frequency of the IHH induction. There were significant differences in Cygb expression between IHH2x and IHH3x groups compared to normoxia group, and between IHH1x, IHH2x and IHH3x compared to AHH group. There were significant differences in Ngb expression between IHH2x and IHH3x groups compared to normoxia group and between IHH2x and IHH3x groups compared to AHH group. The specific activity of AChE was increased significantly since the first induction of AHH, but then decreased in IHH3x. There were significant differences in the specific activity of AChE between IHH2x and IHH3x groups compared to normoxia and between IHH2x and IHH3x groups compared to IHH1x groups. Conclusions: We conclude that IHH, especially IHH3x, seems to induce the protective adaptive response in the rat brain tissue through the changes of these three parameters.

Trypanosoma evansi is a Hemoflagella parasite that infects cattle and is known as the agents of Surra. Several other trypanosome species infects mammals: T. equiperdum, T. b. rhodesiense, T. b. gambiense, T. vivax, T. congolense, T.theileri . Some of these species is quite difficult to be distinguished morphologically with T. evansi through conventional techniques (thin blood smear). Molecular technique by polymerase chain reaction (PCR) is reported to have the ability to identify, characterize and diagnose trypanosomes accurately. However, a single PCR used is relatively expensive because it takes at least two or more pairs of primers to determine T. evansi . The purpose of this study is to develop T. evansi species identification techniques by multiplex PCR/mPCR (the three pairs of primer in one reaction) that takes the relatively fast and inexpensive. A total of 31 isolates T.evansi were obtained from Bblitvet Culture Collection (BCC) and the Department of Parasitology BBLitvet used in this study. Isolates represent isolates from endemic areas and Surra outbrake isolated from 1988-2014. DNA extraction performed on each sample, including Bang 87 isolates which has been purified as a positive control. Primers used are specific for T. evansi , the ITS-1, Ro Tat 1.2 VSG and ESAG 6/7. Before running mPCR, each primer is optimized by using a single PCR. The results showed that the three primers can be combined in a single reaction with mPCR technique and amplify each DNA fragment target perfectly, so identified 31 isolates as T. evansi . This technique can be applied in the field with a lower cost and faster time. Key Words : Trypanosoma evansi , Identification, Multiplex PCR

Objective: Glioma is the most common human primary brain tumor which is highly resistant to oxidative stress-based anticancer. The aim of this study was to analyze the effect of rotenone-induced reactive oxygen species (ROS) on the modulation of manganese superoxide dismutase (MnSOD) expression and cell viability in human glioblastoma (GBM) T98G cells.Methods: In this in vitro experimental study, T98G cells were treated with high-dose rotenone (0.5, 5, and 50 μM, respectively). Following rotenone treatment and intracellular ROS, both peroxide and superoxide radicals were determined. Moreover, we analyzed MnSOD mRNA expression, protein, and specific activity, as well as cell survival including viability, proliferation, apoptosis, and mitochondrial structure.Results: High-dose rotenone treatment of T98G cells significantly increased intracellular ROS and MnSOD mRNA, but its protein and specific activity definitely decreased. The treatment also led to a reduction of cell viability, enhancement of apoptosis, and disruption of mitochondrial integrity.Conclusion: Overproduction of ROS in rotenone-treated human GBM T98G cells could suppress the MnSOD protein level and activity even though mRNA synthesis has been increased. This modulation led to reduced survival of T98G cells through induction of cell death rather than inhibition of cell proliferation.

The aim of this study was to evaluate the effects of methanol extract of Jatropha multifida leaves on the wound healing process and to investigate the wound healing activity based on reduced numbers of PMN (polymorpho nuclear) leukocytes and increased numbers of fibroblasts. Methanol extract of dried leaves of J. multifida was used in the wound healing activity studies. The study subjects were 36 white male Sprague Dawlay rats aged 2 months with 150-200 gram body weight. The subjects were divided into 4 groups and experimentally injured: Group I (negative control) underwent injury without subsequent treatment; group II (positive control) received topical treatment with Bethasone-N after injury; group III (solvent control) was treated with 70% methanol; group IV (treatment group) was treated with 10 mg methanol extract of J. multifida Each group consisted of 3 rats, which were decapitated on days 3, 6, and 13 after the start of treatment. Histological preparation was stained with hematoxyline-eosin (HE) and was continuously examined by counting the numbers of PMN leukocytes and fibroblasts as indicators of wound healing on days 3, 6, and 13 of treatment. The study showed lower numbers of PMN leukocytes in subjects treated with the extract of J. multifida as compared to the other groups. The numbers of fibroblasts were significantly higher on days 6 and 13 of treatment. The treatment of injuries with methanol extract of leaves from J. multifida provided better results compared to the other groups in our study.