In vitro and in silico analysis of phytochemical compounds of 96% ethanol extract of semanggi (Marsilea crenata Presl.) leaves as a bone formation agent.

OBJECTIVES: Osteoporosis is the result of an imbalance in the rate of bone resorption and bone formation due to a decrease in estrogen. Phytoestrogens are plant compounds with structures and functions similar to estrogen. Phytoestrogens that bind to estrogen receptors in bone cells are able to modulate bone formation. Semanggi (Marsilea crenata Presl.) is a plant that contains phytoestrogens. The purpose of this study was to observe the expression of osteocalcin and predict the content of extract phytoestrogens through a computer simulation study to study the bone formation activity of the 96% ethanol extract of M. crenata leaves on hFOB 1.19 cells. METHODS: hFOB 1.19 cells were cultured in 24-well microplates, and 96% ethanol extract of M. crenata Presl. leaves was added at 62.5, 125 and 250 ppm. The expression of osteocalcin was analyzed using CLSM immunocytochemistry. Using PyRx 0.8 software and 1ERE protein for molecular docking, the compound was analyzed by computer. RESULTS: The 96% ethanol extract of M. crenata Presl. leaves can increase the expression of osteocalcin, the optimal dose is 125 ppm, and p<0.05 is 881.658 AU. In silico study was obtained six compounds that showed similar activity 17ß-estradiol as ER-ß agonists. CONCLUSIONS: The 96% ethanol extract of M. crenata Presl. leaves contain six compounds that are thought to be phytoestrogens and ER-ß agonists, and play a role in increasing bone formation activity and have the potential to be used as an oral drug.

Prediction of compounds with antiosteoporosis activity in Chrysophyllum cainito L. leaves through in silico approach.

OBJECTIVES: Estrogen deficiency causes various health problems in postmenopausal women, including osteoporosis. Phytoestrogen emerged as a potential alternative of estrogen with minimum side effects. The aims of this study were to analyze the metabolite profiling results of various extract of Chyrsophyllum cainito L. leaves, which contain phytoestrogen, through in silico study against 3OLS protein, an X-ray protein of ERß, so it can predict the types of the phytoestrogen contents which have antiosteoporosis property. METHODS: In silico analysis was carried out for the compounds from the metabolite profiling data of C. cainito leaves from our previous study. The structure compounds from metabolite profiling results of various extract of C. cainito leaves were prepared with Avogadro 1.0.1 software, molecular docking was done using PyRx 0.8 software, and Biovia Discovery Studio Visualizer 2016 software was used to visualize the structure of compounds against 3OLS protein. The physicochemical characteristics of the compounds were analyzed using the SwissADME web tool. RESULTS: From in silico studies, it was known that there were total 11 compounds in C. cainito leaves that predicted as phytoestrogens which have ERß agonist properties against 3OLS protein. The ERß agonist was a compound that has parameters similar to 17ß-estradiol in its interaction with 3OLS protein, which has a pharmacophore distance of 10.862 Å, and binding to amino acids His 475 and Glu 305 or Arg 346 at receptor-ligand docking simulation. CONCLUSIONS: C. cainito leaves contain 11 compounds that are predicted to be phytoestrogens with ERß agonist properties, which is responsible for antiosteoporosis activity.

COVID-19 pandemic in Indonesia: Situation and challenges of rehabilitation medicine in Indonesia.

AbstrakCOVID-19 telah menjadi pandemik di Indonesia sejak ditemukannya kasus pertama pada tanggal 2 Maret 2020 di Depok. Peningkatan kasus perhari semakin tinggi sejak akhir Agustus 2020 yang mencapai lebih dari 2000 kasus per hari. Sistem kesehatan di Indonesia perlu ditingkatkan dalam hal kapasitas, termasuk rehabilitasi medik yang harus dilibatkan dari fase akut hingga jangka panjang dalam penanganan pasien COVID-19. Rehabilitasi medik juga diperlukan untuk pasien lain yang bukan COVID-19. Pentingnya keterlibatan, pelayanan rehabilitasi medik dan implementasinya dimasa pandemic COVID-19 memerlukan strategi tersendiri yang harus dilakukan baik oleh pekerja kesehatannya, rumah sakit dan kebijakan pemerintah. Hal ini diperlukan untuk percepatan peningkatan kesehatan pasien, percepatan pemulangan dan menghindari readmisi pasien, dan juga pengoptimalan program kembali bekerja untuk pasien yang sembuh dari COVID-19.AbstractCOVID-19 has become a pandemic in Indonesia since the first cases have been positively diagnosed on 2 March 2020 in Depok. The cases have been increased gradually since the end of August 2020 that has reached 1000 cases per day. The health system in Indonesia needs to be improved in terms of capacity, including rehabilitation medicine that should be involved in all health phases (from acute to long-term) in managing patients with COVID-19. Rehabilitation is also still needed for other non-COVID-19 patients. The importance of involvement and implementation of rehabilitation services during the COVID-19 pandemic will need special strategies that should be done by rehabilitation professionals, hospitals, and government. These are necessary to accelerate the improvement of patients' health, discharge, and avoid re-admission, as well as optimize return-to-work for patients who are recovered from COVID-19.

The enhancement of Arg1 and activated ERß expression in microglia HMC3 by induction of 96% ethanol extract of Marsilea crenata Presl. leaves.

Background Phytoestrogens have a high potential to overcome the neuroinflammation caused by estrogen deficiency. Marsilea crenata Presl. is a plant known to contain phytoestrogens. This research aimed to report the activity of a 96% ethanol extract of M. crenata leaves in inducing activation of microglia HMC3 cell to M2 polarity, which has anti-inflammatory characteristics. Methods The study was done by culturing microglia HMC3 cell in 24-well microplate and inducing it with IFN-γ for 24 h to activate the cell to M1 polarity, which has proinflammatory characteristics. The 96% ethanol extract was added with various doses of 62.5, 125, and 250 ppm. Genistein, 50 µM, was used as a positive control. The analysis of the immunofluorescence of Arginase-1 (Arg1) and ERß as markers was done using a convocal laser scanning microscope. Results The result of Arg1 shows a significant difference in Arg1 expression in the microglia HMC3 cell line between the negative control and all treatment groups at p < 0.05, with the best result at 250 ppm, whereas for ERß, the results show, at doses of 125 and 250 ppm, that the 96% ethanol extract of M. crenata leaves decrease the activated ERß expression at p < 0.05, with the best result at 250 ppm. The Arg1 and activated ERß expression have a weak negative relationship with the Pearson correlation test. Conclusions The 96% ethanol extract of M. crenata leaves has an antineuroinflammation activity through the induction of Arg1 and activated ERß expression in microglia HMC3 cell, with the best dose at 250 ppm.

Antineuroinflammation activity of n-butanol fraction of Marsilea crenata Presl. in microglia HMC3 cell line.

Background Neuroinflammation is one of the main causes of neurodegenerative events. Phytoestrogen is a group compounds that have an estrogen-like structure or function. Phytoestrogen has a high potential to overcome neuroinflammation caused by estrogen deficiency in postmenopausal women. Marsilea crenata Presl. is a plant known to contain phytoestrogens. This research aimed to analyze the activity of an n-butanol fraction of M. crenata leaves in inhibiting the classical pathway activation of microglia HMC3 cell line to M1 polarity, which has proinflammatory characteristics. Methods Microglia HMC3 cell line was cultured in Eagle's minimum essential medium and induced with IFN-γ for 24 h to activate the cell to M1 polarity in 24-well microplates. The n-butanol fraction was added with various doses of 62.5, 125, and 250 ppm and genistein 50 µM as a positive control. The expression of major histocompatibility complex II (MHC II) as a marker was tested using a confocal laser scanning microscope. Results The result of MHC II measurement shows a significant difference in the MHC II expression in the microglia HMC3 cell line between the negative control and all treatment groups at p<0.05, indicating a non-monotonic dose-response profile. Conclusions The best dosage to inhibit MHC II expression was 250 ppm with the value of 200.983 AU. It is then concluded that n-butanol fraction of M. crenata leaves has antineuroinflammation activity due to its phytoestrogens.

Brain Derived Neurotropic Factors in Speed vs. Inclined Treadmill in Young Adult Healthy Male With Occult Balance Disorder.

Background: There is an increase in fall risk among elders and young adults consecutively due to various causes. Occult balance disorder may be among the abnormal causes of falling in young adults as well as elders. The One Leg Stance (OLS) test is used to diagnose this balance performance; it's a proven test to measure static balance function which would lead to dynamic balance function. It has been proven that aside from cardiopulmonary exercises, treadmill workout can be used as a dynamic balance exercise. The Brain Derived Neurotropic Factor (BDNF) increases balance function through the treadmill exercise (the inclination and speed). This hormone is one of the tropical hormones generated in neurons, muscles, hematopoietic tissue and it is characterized by neurons morphology regulation and neuroplasticity. Materials and Methods: We divided 20 healthy young adult men to work out on inclination and speed groups treadmill exercise. The workout lasted for 2 weeks. We immediately observed the effect of exercise on serum BDNF as two tests were taken on before and 30 min after the workout. Result: There were significant increases of pre-exercise serum BDNF level in speed group between the first and the last exercise (p = 0.001), post-exercise between the first day and the last exercise (p = 0.001). No significant increase of serum BDNF in speed group pre- and post-exercise on the first exercise (p = 0.159), pre- and post-exercise on the last exercise (p = 0.892). There was no significant increase in serum BDNF in inclination group on all parameters (p > 0.05). The serum BDNF is actually a neurotropic factor that affects not just the neuronal system, but also molecular energy and metabolism regulation. This serum is dependent on the aerobic capacity, lactate production, muscle calcium uptake, and muscle fiber type used in exercises. Furthermore, the serum BDNF is increased by treadmill exercises in escalated speed. Conclusion: Treadmill exercises with average speed escalation increase the serum BDNF.

Low-Intensity Exercise with Blood Flow Restriction Increases Muscle Strength without Altering hsCRP and Fibrinogen Levels in Healthy Subjects.

Strengthening exercise combined with blood flow restriction potentially increases muscle strength. This type of exercise does not require heavy weight liftings and is a feasible method to be performed by persons suffering illnesses. However, strengthening exercise may induce inflammatory responses due to muscle and vascular endothelial damage. This study aimed to investigate alterations of high-sensitivity C-reactive protein (hsCRP) and fibrinogen levels in healthy subjects after five weeks of low intensity resistance training (LIRT) with blood flow restriction (BFR) on increasing strength in comparison with high intensity resistance training (HIRT) and LIRT alone, and to evaluate aspects related to the relative safety of LIRT + BFR. Eighteen healthy subjects were randomized into 3 groups. The HIRT group: 70% of One-Repetition Maximum (1-RM); LIRT + BFR group: 30% of 1-RM with BFR (a modified 13-cm wide cuff was used); LIRT group: 30% of 1-RM. The peak torque of isokinetic contraction of the left elbow flexor in each subject was measured before and after 5 weeks of resistance exercises to determine any increases in the left biceps brachii muscle strength. Blood markers of homeostasis (fibrinogen) and inflammation (hsCRP) were also measured before and after five weeks of training. Significant increases of strength were demonstrated between the five weeks of resistance exercises in the HIRT group (P = 0.003) and the LIRT + BFR group (P = 0.001). Peak torque of isokinetic contraction of the left flexor elbow joint at 60° per second angular velocity showed that the LIRT + BFR group produced the greatest peak torque increase than the HIRT group. There were no significant changes in the hsCRP levels in all the groups (P > 0.05) after five weeks of intervention. No significant differences of fibrinogen levels were found in the HIRT group (P = 0.500) and the LIRT + BFR group (P = 0.405), but significant decreases were found in the fibrinogen levels in the LIRT group (P = 0.017). The LIRT + BFR increases in the muscle strength were as significant as in HIRT without altering the fibrinogen and hsCRP levels in the healthy subjects. In this study, LIRT + BFR showed increase muscle strength without any vascular problems.

Pain Relief with Wet Cupping Therapy in Rats is Mediated by Heat Shock Protein 70 and ß-Endorphin.

BACKGROUND: Wet cupping therapy is a complementary therapy in pain management. The mechanism of this therapy, however, needs further elucidation. Cells injured by wet cupping therapy seem to stimulate the expression of heat shock protein 70 (HSP70). Its benefit in pain reduction could be mediated by the expression of ß-endorphin. This study aimed at determining the correlation between HSP70 and ß-endorphin after wet cupping therapy. METHODS: Sixteen male Wistar rats were divided into control (CG; n=8) and treatment (TG; n=8) groups. The rats in both groups were injected with complete Freund's adjuvant (CFA) at the footpad. In the TG, wet cupping therapy was done at the left and right paralumbar regions 48 hours after the CFA injection. Twenty-four hours after therapy, the hot plate test was done to assess pain threshold. Thereafter, immunohistochemistry from the skin subjected to wet cupping therapy was conducted for HSP70 and ß-endorphin. RESULTS: The expression of HSP70 was significantly higher in the keratinocytes of the TG (20.25±3.53; P<0.001) than in the keratinocytes of the CG (10.50±2.44; P<0.001). The expression of ß-endorphin was significantly higher in the keratinocytes of the TG (22.37±3.52; P<0.001) than in the keratinocytes of the CG (5.12±1.72; P<0.001). The results also revealed a high correlation between HSP70 and ß-endorphin (ß=0.864; P<0.001). Pain threshold after wet cupping therapy was significantly higher in the TG (22.81±6.34 s; P=0.003) than in the CG (11.78±3.56 s). CONCLUSIONS: The benefit of wet cupping therapy in terms of pain reduction in rats could be mediated by the expression of HSP70 and ß-endorphin.