Deteksi populasi Cd3+Cd4+Cd25+Foxp3+ T-regulator pada Limpa Mencit Galur Balb/c dan Swiss-webster yang Mudah dan Cepat dengan Metode Flow Cytometry

Sel T-regulator (Treg) memainkan peran kunci dalam mengendalikan autoimunitas, respon alergi, peradangan, dan respons terhadap infeksi. Sel T Cd4+ yang secara konstitutif mengekspresikan Cd25 dan protein forkhead 3 (Foxp3; yaitu, Cd3+Cd4+Cd25+Foxp3+) atau disebut juga sebagai Treg fungsional, telah terbukti memainkan peran utama dalam pemeliharaan toleransi dan homeostasis imun, sehingga deteksi Treg fungsional sangat penting dilakukan sebagai salah satu parameter imunomodulasi. Penelitian ini bertujuan untuk mendeteksi populasi sel Treg (Cd3+Cd4+Cd25+Foxp3+) pada organ limpa mencit galur Balb/c dan Swiss-webster dengan menggunakan metode yang mudah, cepat, serta terjangkau. Metode deteksi sel Treg dilakukan dengan cara isolasi organ limpa mencit Balb/c dan Swiss-webster kemudian dilakukan preparasi dan deteksi sel menggunakan instrumen flow cytometry. Hasil populasi sel Treg yang didapatkan berturut-turut pada mencit Balb/c dan Swiss-webster adalah 2,02 ± 0,36% dan 3,14 ± 1,64% dari total populasi yang dideteksi dengan hanya membutuhkan waktu 145 menit sejak pembedahan. Berdasarkan hasil tersebut dapat disimpulkan bahwa pengamatan populasi sel Treg pada splenosit mencit galur Balb/c dan Swiss-webster dengan flow cytometry dapat dilakukan dengan mudah dan praktis dengan pewarnaan kombinasi antibodi yang tertera pada manuskrip ini. Selain itu, metode pewarnaan yang tertera dapat digunakan sebagai salah satu acuan untuk mendeteksi imunomodulator pada berbagai model hewan coba yang menggunakan mencit galur Balb/c dan Swiss-webster, khususnya yang berhubungan dengan populasi sel Treg

2-(3(chloromethyl)benzoyloxy)benzoic Acid Increases CD4+ Regulatory T-Cell Population and FoxP3 Expression in Lipopolysaccharide-induced Mice

ACKGROUND: Lipopolysaccharide (LPS) has been reported to increase CD4+ regulatory T-cell (CD4+ Treg) populations. Acetylsalicylic acid (ASA) has been reported to have immunomodulatory activity, but it may induce chronic gastric ulceration. Another salicylic acid-bearing compound, 2-(3-(chloromethyl) benzoyloxy)benzoic acid (3-CH2Cl), has been reported to have less gastric mucosal damage. However, the effect of 3-CH2Cl on CD4+ Tregs in LPS-induced mice is still unknown. Therefore, the present study was conducted to investigate the immunomodulatory effect of 3-CH2Cl on CD4+ T-cell and CD4+ Treg populations as well as FoxP3 expression in LPS-induced mice. METHODS: Synthesis of 3-CH2Cl was performed by mixing salicylic acid and chloromethylbenzoylchloride with the catalyzation of pyridine, acetone and heat. The 3-CH2Cl tablets were prepared using direct compression method. After intraperitoneal injection of 1 mg/kg BW LPS to mice, 60 mg/kg BW ASA or 60 mg/kg BW 3-CH2Cl was given Abstract orally for 3 days. The splenocyte was obtained through splenectomy and collagenase digestion. The population of CD4+ T-cells and CD4+ Tregs, as well as the splenic FoxP3 expression were determined using flow cytometry technique. RESULTS: CD4+ T-cell populations in mice treated with LPS and 3-CH2Cl or ASA were lower than those treated with LPS merely. Meanwhile, CD4+ Treg populations and FoxP3 expression levels in mice treated with LPS and 3-CH2Cl or ASA were higher than those treated with LPS merely. CONCLUSION: Since 3-CH2Cl could decrease CD4+ T-cell population and increase CD4+ Treg population mediated by the increase of FoxP3 expression in LPSinduced inflammation, it may act as a potential therapeutic drug to reduce inflammatory conditions

Deteksi populasi Cd3+Cd4+Cd25+Foxp3+ T-regulator pada Limpa Mencit Galur Balb/c dan Swiss-Url menuju focus and scope:ster yang Mudah dan Cepat dengan Metode Flow Cytometry

Sel T-regulator (Treg) memainkan peran kunci dalam mengendalikan autoimunitas, respon alergi, peradangan, dan respons terhadap infeksi. Sel T Cd4+ yang secara konstitutif mengekspresikan Cd25 dan protein forkhead 3 (Foxp3; yaitu, Cd3+Cd4+Cd25+Foxp3+) atau disebut juga sebagai Treg fungsional, telah terbukti memainkan peran utama dalam pemeliharaan toleransi dan homeostasis imun, sehingga deteksi Treg fungsional sangat penting dilakukan sebagai salah satu parameter imunomodulasi. Penelitian ini bertujuan untuk mendeteksi populasi sel Treg (Cd3+Cd4+Cd25+Foxp3+) pada organ limpa mencit galur Balb/c dan Swiss-webster dengan menggunakan metode yang mudah, cepat, serta terjangkau. Metode deteksi sel Treg dilakukan dengan cara isolasi organ limpa mencit Balb/c dan Swiss-webster kemudian dilakukan preparasi dan deteksi sel menggunakan instrumen flow cytometry. Hasil populasi sel Treg yang didapatkan berturut-turut pada mencit Balb/c dan Swiss-webster adalah 2,02 ± 0,36% dan 3,14 ± 1,64% dari total populasi yang dideteksi dengan hanya membutuhkan waktu 145 menit sejak pembedahan. Berdasarkan hasil tersebut dapat disimpulkan bahwa pengamatan populasi sel Treg pada splenosit mencit galur Balb/c dan Swiss-webster dengan flow cytometry dapat dilakukan dengan mudah dan praktis dengan pewarnaan kombinasi antibodi yang tertera pada manuskrip ini. Selain itu, metode pewarnaan yang tertera dapat digunakan sebagai salah satu acuan untuk mendeteksi imunomodulator pada berbagai model hewan coba yang menggunakan mencit galur Balb/c dan Swiss-webster, khususnya yang berhubungan dengan populasi sel Treg

Pre-meal high-performance inulin supplementation reduce post-prandial glycaemic response in healthy subjects: A repeated single-arm clinical trial

Background and aims: High-performance (HP) inulin, a dietary fiber consists of more than 10 fructose polymers, have been shown to reduce post-prandial glycaemic response (PPGR) and could prevent the occurrence of Type-2 diabetes mellitus (T2DM). Currently, there are no data on whether pre-meal HP inulin supplementation could decrease PPGR. Methods: 8 healthy adults consumed 20 g of formula that contain 60.2% inulin (w/w) dissolved in water. Blood glucose was measured in fasted participants and at 30e120 min after starting to eat a prepared meal. This test was repeated every week with different supplement formulas. Conclusion: pre-meal HP Inulin formula supplementation could suppress the post-prandial glycaemic response

Uji Aktivitas Analgesik Tablet Asam 4-(Klorometil) Salisilat pada Mencit Putih dengan Metode Writhing Test dan Hotplate

Acetylsalicylic acid is an analgesic drug that still popularly used. However, acetylsalicylic acid may result undesirable side effect of gastrointestinal as it can cause gastric ulcer to bleeding. Previous research has succeeded synthesizing new compound derived from salicylic acid, namely 4-(chloromethyl)salicylic acid which has the potential to reduce its side effects with a better pharmacological effect than acetylsalicylic acid. In its development, the formulation of the compound in tablet dosage form has been carried out. However, until now there has not been a pharmacological test to determine whether it still provides a greater effect than acetylsalicylic acid after being formulated. This study is to test the analgesic activity of 4-(chloromethyl)salicylic acid tablets in albino mice using writhing test and hotplate methods. Analgesic activity test using hotplate method was carried out on albino mice which given the suspension of the tablet sample and the observations were made in a periodic of time. Analgesic activity test using writhing test method was carried out in 0.6% acetic acid-induced albino mice, where the suspension of the tablet was administered 30 minutes before administration of acetic acid. Analgesic activity was assessed by counting the number of writhing and the results were statistically compared with a one-way ANOVA. Tests were also performed on acetylsalicylic acid as a positive control and suspension as a negative control group. There were significant differences (P<0.05) of the analgesic activity observed among groups. Both methods show that 4-(chloromethyl)salicylic acid tablets provide stronger analgesic activity than acetylsalicylic acid

Tablet Formulation of 2-(3-(Chloromethyl) benzoyl) oxy) benzoic Acid by Linear and Quadratic Models

Purpose: This research determines the effect of sodium lauryl sulfate (SLS) as a surfactant, croscarmellose sodium (CS) as a disintegrating agent, and SLS–CS combinations on 2-((3-(chloromethyl)benzoyl)oxy)benzoic acid (3CH2Cl) (log P = 3.73) tablet formulations. In addition, this study aims to determine the optimum of the 3CH2Cl tablet formula. Methods: The tablets are manufactured through direct compression according to the simplex lattice design. The optimal SLS and CS concentration was determined in vitro using linear and quadratic models to achieve better tablet disintegration and dissolution. Results: The same linear and quadratic coefficient profiles of SLS and CS indicate that the combined coefficient of SLS–CS with a quadratic model can be used to predict the effect of the SLS–CS combination. Based on the linear model coefficients, SLS and CS increase the value of flow time (9.35; 7.65), Carr index (26.17; 21.17), hardness (9.84; 7.44), friability (0.38; 0.31), disintegrating time (5.74; 2.62), and drug release (84.28; 58.65). The quadratic model coefficient shows that SLS–CS combinations increase flow time (0.60), Carr index (2.00), hardness (1.00), and disintegrating time (1.04). Meanwhile, they decrease friability (−0.02) and drug release (−9.10). Conclusions: SLS, CS, and SLS–CS combinations affect the quality of tablet mass and tablets. The optimum tablet formula was 3CH2Cl (300 mg), Ne (9.38%), SLS (0.92%), CS (2.33%), MCC (5%), and SDL (ad 800 mg). 3CH2Cl has analgesic activity despite the presence of tablet excipients. The 3CH2Cl tablet is an innovative formulation and a new alternative for future analgesic drugs

Ethanolic extract Ocimum sanctum Linn. induces an apoptosis in human lung adenocarcinoma (A549) cells

Ocimum sanctum (OS) is tropical herbal plant which is easy to find and widely used as a vegetable food in Indonesia. In last decade, lung adenocarcinoma was in top position as male cancer disease in Indonesia. Recently, emerging data showing the extracts of different species of Ocimum exhibiting the anti-tumor properties. Further studies on lung lewis carcinoma demonstrated pro-apoptosis effects after the treatment with Ocimum extracts. However, the effect of OS of Indonesian origin in human alveolar pulmonary adenocarcinoma A549 cells remain unclear. Therefore, we aimed to investigate effects of ethanolic extract OS (EEOS) in A549 cell culture systems. Cell adhesion and viability assays revealed that EEOS significantly decreased the attachment into extracellular matrix of A549 cells. Morphological examination AO/EB and DAPI staining indicated that EEOS induced the cells shrinkage, DNA fragmentation and condensation of A549 cells. Further, EEOS treatment induced the apoptosis rate followed by up-regulation of reactive oxygen species (ROS), caspase-3 expression and decreased antiapoptotic protein Bcl-2. This condition also suppressed the expression of SOD2 as well as the GPx. In conclusion, our findings indicate that EEOS suppressed the viability of A549 cells, which may result from the activation of ROS promoting the apoptosis signaling via mitochondrial intrinsic pathway. Taken together, EEOS might be a good therapeutic potential to further understand its properties in the treatment of lung carcinom

Choline Transporter-Like Protein-2: New von Willebrand Factor-Binding Partner Involved in Antibody-Mediated Neutrophil Activation and Transfusion-Related Acute Lung Injury

In contrast to other antibodies involved in transfusion-related acute lung injury, anti-HNA-3a antibodies are incapable of inducing direct neutrophil activation and seem to interact with endothelial cells (ECs) primarily. In animal studies, anti-HNA-3a-mediated transfusion-related acute lung injury could be precipitated in the absence of neutrophils, but was stronger when neutrophils were present. In a different context the target protein of these antibodies, choline transporter-like protein-2 (CTL-2), was reported to interact with a protein of the inner ear carrying 2 von Willebrand factor (VWF) A-domains. These observations prompted us to investigate whether VWF might be involved in anti-HNA-3a-mediated neutrophil activation, and whether signaling via CD11b/CD18 is involved, as in various other experimental settings.status: publishe

Managing spondylitis tuberculosis in a patient with underlying diabetes and hypothyroidism: A case report

Tuberculosis (TB) remains one of the highest Asia’s health problems. Spondylitis TB in diabetes mellitus (DM) and hypothyroidism patients is a rare case of extrapulmonary tuberculosis. However, there is a lack of therapeutic guidelines to treat spondylitis TB, particularly with type 2 DM (T2DM) and hypothyroidism as comorbidities. Here we present a case of spondylitis TB with T2DM and hypothyroidism in a relatively young patient and its therapeutic procedure. We report the case of a 35-year-old male patient from Surabaya, Indonesia. Based on anamnesis, physical examination, and magnetic resonance imaging, the patient has been categorized in stage II of spondylitis TB with grade 1 paraplegia. Surprisingly, the patient also had a high HbA1c level, high thyroid stimulating hormone, and low free T4 (FT4), which indicated T2DM and hypothyroidism. A granulomatous process was observed in the histopathological section. The antituberculosis drugs isoniazid and rifampicin were given. In addition, insulin, empagliflozin, and linagliptin were given to control hyperglycemia conditions, and also levothyroxine to control hypothyroidism. CONCLUSION The outcome was satisfactory. The patient was able to do daily activities without pain and maintained normal glycemic and thyroid levels. For such cases, we recommend the treatment of spondylitis TB by spinal surgery, together with T2DM and hypothyroidism therapies, to improve the patients’ condition. Prompt early and non-invasive diagnoses and therapy are necessary

Ethanolic extract Ocimum sanctum Linn. induces an apoptosis in human lung adenocarcinoma (A549) cells

Ocimum sanctum (OS) is tropical herbal plant which is easy to find and widely used as a vegetable food in Indonesia. In last decade, lung adenocarcinoma was in top position as male cancer disease in Indonesia. Recently, emerging data showing the extracts of different species of Ocimum exhibiting the anti-tumor properties. Further studies on lung lewis carcinoma demonstrated pro-apoptosis effects after the treatment with Ocimum extracts. However, the effect of OS of Indonesian origin in human alveolar pulmonary adenocarcinoma A549 cells remain unclear. Therefore, we aimed to investigate effects of ethanolic extract OS (EEOS) in A549 cell culture systems. Cell adhesion and viability assays revealed that EEOS significantly decreased the attachment into extracellular matrix of A549 cells. Morphological examination AO/EB and DAPI staining indicated that EEOS induced the cells shrinkage, DNA fragmentation and condensation of A549 cells. Further, EEOS treatment induced the apoptosis rate followed by up-regulation of reactive oxygen species (ROS), caspase-3 expression and decreased antiapoptotic protein Bcl-2. This condition also suppressed the expression of SOD2 as well as the GPx. In conclusion, our findings indicate that EEOS suppressed the viability of A549 cells, which may result from the activation of ROS promoting the apoptosis signaling via mitochondrial intrinsic pathway. Taken together, EEOS might be a good therapeutic potential to further understand its properties in the treatment of lung carcinom