Secreted Left Atrial P-Selectin in Mitral Stenosis after PBMV: When to Measure

The glycoprotein P-selectin, a membrane component of cell storage granules, is rapidly translocated from ?-granules of platelets and the Weibel- Palade bodies of endothelial cells to the cell surface following an inflammatory process or other stimulations. P-selectin is a cell adhesion molecule of activated platelets and endothelial cells of interest because of its role in modulating interactions between blood cells and the endothelium, and also because of the possible use of its soluble form in plasma as a predictor of adverse cardiovascular events.1 In endothelial cells, within minutes of its stimulation in vitro by inflammatory mediators, such as histamine, thrombin, or phorbol esters, or hypoxia, WeibelPalade bodies are mobilized and their von Willebrand factor are degranulated. At the same time, P-selectin is also expressed at the surface as quick astwo minutes after stimulation. However, this expressionis short-lived, reaching its peak after only 10 minutes, declining back to baseline levels after 3 hours. Additional synthesis of P-selectin is brought about within 2 hours by cytokines, such as interleukin-1 (IL-1), tumor necrosis factor-? (TNF-?), and by thrombin, lipopolysaccharide or oxygen radicals. Immunofluorescence and confocal laser cytometry are usually used to measure the translocation upon this activation.

Pemeriksaan BNP atau NT pro BNP pada Pasien Gagal Jantung

Beberapa permasalahan masih sering dihadapi berkaitan dengan diagnosis, evaluasi keberhasilan terapi, penekanan angka morbiditas dan mortalitas gagal jantung. Seperti misalnya, pada kondisi gawat darurat yang membutuhkan diagnosis dan terapi cepat dan akurat, kadang sulit untuk membedakan sesak nafas akibat gagal jantung atau akibat penyakit lain seperti penyakit paru dan metabolik. Hingga saat inipun pemulangan pasien dari perawatan rumah sakit sebagian besar didasarkan atas berkurangnya atau hilangnya keluhan, meskipun mungkin belum dilakukan titrasi terapi yang optimal. Hal itu menyebabkan tingginya angka kematian dan rawat ulang pasien gagal jantung. Lebih jauh, hingga saat ini belum ada metoda yang rutin dipakai untuk meng-evaluasi efek dari terapi yang telah diberikan pada level molekuler. Oleh karena itu, perlu ada alat diagnostik untuk mengetahui fungsi jantung secara cepat dan tepat, yang dapat membedakan sesak karena gagal jantung atau faktor lain, dan dapat pula menjadi predikor prognosis pasien gagal jantung

The Effect Of Red Rosella Tea (Hibiscus sabdariffaLinn) on the Inhibition of NF-?B Activation, TNF-aand ICAM-1- Protein Expressions Following Treatment with Ox-LDL in HUVECs

Background. Atherosclerosis disease has been a global threat. Ox-LDL was one of major source of atherogenesis process, through the ROS formation which would then activate NF-KB and increase the expression of TNF-a and ICAM-1, leading to the occurrence of inflammation. Recent studies indicate that red Rosella tea is a tropical plant containing antioxidant (protocatechuic acid and antocyanin) and ascorbate acid. This tea is widely distributed in Indonesia. The mechanism of action of this tea on the inhibition of atherogenesis has not been largely known. This study aim to investigate the effect and mechanism of the red Rosella tea on the expression of NF-KB, TNF-aand ICAM-1. Methods and results.Cultured endothelial cell, HUVECs (Human Umbilical Vein Endothelial), is used for a model of endothelial cells. There are 5 groups: 1) HUVECs culture without Ox-LDL as negative control; 2) HUVECs culture with Ox-LDL as positive control; and 3 groups given various dosages of the red Rosella tea (0.001 mg/ml, 0.005 mg/ml and 0.01 mg/ml) and Ox-LDL. OxLDL (40 mg/ml) is added on HUVECs to stimulate responds of endothelial cells mimicking dyslipidemia condition. The Red Rosella tea were given 2 h before Ox-LDL treatment. The NF-KB activaton, the protein expressions of TNF-aand ICAM-1 are detected by immunohistochemistry. Treatment with red rosella tea on Ox-LDL-treated HUVECs resulted in prevention of NF-KB activation which in turn inhibited the increased of TNF-aand ICAM-1 protein expressions. Spearmans correlation analysis showed that there is negative correlations between dosages of red tea with NF-KB activation, TNF-aand ICAM-1 protein expressions (p=0.01, Correlation Coeff= -1). Conclusions. This study indicates the benefit of red Rosella tea on the prevention and medication for atherosclerosis disease

Efek Ekstrak Daun Kelor (Moringa Oleifera) Dalam Menghambat Aktifasi NFkB, Ekspresi TNF-adan ICAM-1 pada HUVECS yang Dipapar LDL Teroksidasi

Background. Atherosclerosis is a chronic inflammation process of vascu-lar endothelial cells. Increased oxydized LDL(OxLDL) is one of the most potent inducer of atherogenesis. OxLDL induces an increased ROS (Reac-tive Oxigen Species) and also acts as cytotoxic and chemotaxis factor for monocytes result in accumulation of inflammatory cells. Nuclear Factor Kappa Beta(NFkB) is a transcription factor that plays an important role in this inflammatory proces. NFkB compose of heterodimer molecules of p50 and p65, which bind to its inhibitor, IkB, leading to its inactive form in cytoplasm. OxLDL activates NFkB complex by phosphorylate IkB resulting in released p50-p65-IkB binding and translocation of p50-p65 into the nucleus. p50-p65 then binds to promoter and activates transcription of target genes. NFkB activation therefore increase gene and protein expressions of target molecules such TNF-a, ICAM-1, VICAM, etc. This study aimed to examine whether Moringa oleiferainhibits activation of NFkB dan expression of cytokine TNF-aand adhesion molecule ICAM-1.Methods and results.Human umbilical vein endothelial cells (HUVECs) treated with OxLDL were used as model of atherosclerosis. The increased NFkB activity was measured by indirect method using p50 subcellular lo-calization by immunohistochemistry. 40ug/mL OxLDL, 0.01 gr/mL and 0.005 gr/mL Moringa oleiferawere used based on preliminary study.Conclusions. This study showed that OxLDL significantly induce NFkB activation and increase protein expression of TNF-aand (ICAM-1). This study also observed that Moringa oleiferasignificantly inhibit NFkB activa-tion, and prevent an increased TNF-aand ICAM-1 expression at protein level in OxLDL-treated HUVECs as compare to the controls. Moringa oleifera dose of 0.01mg/mL has a better inhibition effect as compare to that of 0.005 gr/mL.Latar Belakang. Aterosklerosis merupakan keradangan kronis pada sel endotel pembuluh darah. Peningkatan LDL teroksidasi (OxLDL) merupakan salah satu perangsang utama terjadinya aterogenesis. OxLDL menginduksi kenaikan ROS (Reactive Oxigen Species), juga bersifat sitotoksis dan kemotaksis bagi monosit, sehingga mengakibatkan penumpukan sel-sel radang. Nuclear Factor Kappa Beta(NFkB) adalah faktor transkripsi yang mempunyai peran penting terhadap terjadinya keradangan ini. NFkB terdiri dari heterodimer p50 dan p65 yang berikatan dengan inhibitor Kappa B (IkB), sehingga dalam sitoplasma berbentuk inaktif. Dengan cara memfosforilasi IkB, maka OxLDL kompleks NFkB melepas ikatan p50-p65-IkB. Selanjutnya p50-p65 berpindah ke inti sel, dan melekat pada promoter (urutan DNA yang penting untuk memicu terjadinya transkripsi) mengaktifasi proses transkripsi dari gen-gen target. Oleh karena itu, peningkatan aktivitas NFkB akan meningkatkan ekpresi gen dan protein-protein yang menjadi target NF?B antara lain TNF-a, ICAM-1 dan VICAM. Penelitian ini bertujuan untuk menguji apakah daun kelor (Moringa Oleifera) dapat menghambat aktifasi NFkB, sehingga pembentukan sitokin TNF-a dan molekul adesi ICAM-1 terhambat.Metode dan hasil.Human umbilical vein endothelial cells (HUVECs) yang dipapar dengan OxLDL dipakai sebagai model yang menyerupai ateosklerosis. Peningkatan aktifitas NFkB diukur secara tidak langsung dengan cara mendeteksi jumlah p50 mengunakan metoda imunohistokimia. Dosis 40 ug/mL OxLDL serta 0.01 gr/mL dan 0.005 gr/mL ekstrak daun kelor dipakai berdasarkan hasil studi pendahuluan. Kesimpulan.Pemberian OxLDL pada HUVECs meningkatkan aktifitas NF?B dan ekspresi TNF-adan ICAM-1 secara bermakna. Dari penelitian ini juga dibuktikan bahwa, ekstrak daun kelor dapat menghambat aktivasi NF?B dan menurunkan ekspresi TNF-adan ICAM-1, yang diiduksi oleh OxLDL dibandingkan kontrol (tanpa pemberian daun kelor, p<0,01). Dosis ekstrak daun kelor 0.01mg/mL mempunyai efek penghambatan terhadap aktifasi NF?B, penurunan ekspresi TNF-adan ICAM-1, yang lebih baik dibandingkan dengan dosis 0.005 gr/mL

Proteolytic activity of recombinant DegP from Chromohalobacter salexigens BKL5

Background: DegP is a serine protease that specifically cleaves and refolds unfolding proteins in the periplasmic space of the cells. To date, there is no information regarding DegP from halophilic bacteria. Chromohalobacter salexigens BKL5 is a moderately halophilic bacterium that has the ability to grow in a media containing more than 15% salt. Therefore, the objectives of this work were to clone and overexpress DegP-encoding gene from C. salexigens BKL5 and characterize its biochemical properties. Results: DegP-encoding gene was overexpressed in Escherichia coli BL21(DE3) CodonPlus in an active form. SDS-PAGE analysis showed that the molecular weight of the recombinant DegP was 45 kDa. Size-exclusion chromatography analysis suggested that recombinant DegP was present in two multimeric states, hexameric and dodecameric, with molecular weights of 297.9 and 579.12 kDa, respectively. Both conformations were enzymatically active when casein was used as substrate for enzymatic assay. Circular dichroism analysis showed that recombinant DegP was composed of 0.21\u20130.29 helical content, which was comparable to the helical content in the crystal structure of E. coli DegP. The basic/acidic residue ratio of recombinant DegP was 0.56, which was slightly higher than that of DegP from extreme halophiles (average, 0.45) but significantly lower than that of DegP from nonhalophiles (average, 0.94). Conclusions: Recombinant DegP from C. salexigens BKL5 showed proteolytic activity when \u3b2-casein was used as a substrate. In silico analysis indicated that recombinant DegP had characteristics similar to those of halophilic proteins depending on its amino acid composition

Improvement of level MDA and SOD Using of Ganoderma Lucidum as Adjunctive Treatment for Statin Based Therapy in High Risk Patient Based on Framingham Score

It is well known from previous research on the importance of antioxidants to inhibit the progression of coronary heart disease in high-risk patients based on framingham scores. Statins are routinely administered in high-risk patients  based on the Dyslipidemia 2016 ESC guidelines but the effects of adding Ganoderma are known to have antioxidant effects, in patients who have received statins. This study aims to assess the effect of antioxidants through measurement of SOD and MDA in patients given a combination of Ganoderma lucidum with statins.Method: A total of 24 patients with high-risk patients based on the Framingham score > 20% were selected as sample of study. Patients with respondents who had never received statin therapy or ACEi or ARB or CCB or beta blocker, or did not routinely consume the drug for 2 weeks were excluded from this study. The addition of Ganoderma lucidum dose of 3x250 mg was added to statins as the first group and Ganoderma lucidum dose of 3x250 mg was added to the Statin combination of Angiotensin Converting Enzym inhibitors (ACE inhibitors) or Angiotensin Receptor Blockers (ARB) and or Calcium Channel Blockers (CCB) and or Beta blockers as second group for 90 days. SOD and MDA levels were measured before and after the administration of Ganoderma Lucidum with the ELISA method.Result: After the administration of Ganoderma Lucidum with standard therapy, there was a decrease in MDA levels and an increase in SOD levels after 90 days in two groups. Conclusion: The addition of Ganoderma Lucidum to standard therapy can reduce oxidant levels in high-risk patients based on framingham scores. Keyword: Superoxyde-dismutase, Malondyaldehide, Oxidative Stress, Antioxidant, Ganoderma Lucidum, Stati

In silico modelling, regulation of cell viability and anti atherosclerotic effect in macrophage by decaffeinated coffee and green tea extract

Background: The evidence of decaffeinated coffee and green tea extract (DCGTE) in amelioration of atherosclerosis through foam cell formation inhibition has not been established. This study tried to predict the potential role of coffee and tea in foam cell inhibition through in silico modelling and to investigate the effect of DCGTE on the viability and regulation of foam cell inhibition effect in macrophage cell. Methods: Prediction of physicochemical properties of secondary metabolite of coffee and tea was computed with Swiss ADME. Simulation of molecular docking was performed using PyRx Autodock Vina. Prediction of biological activities was done with PASS SERVER and analyzed the suitability with KEGG Pathway: lipid and atherosclerosis. The effect of DGCTE on macrophage viability was assessed with WST-1 assay then synergistic score was calculated with Synergy Finder. The effect of DCGTE in foam cell formation was examined with light microscope after stained with ORO on oxLDL-stimulated Raw264.7. Results: Molecular docking analysis revealed a strong affinity binding between all active compound of tea or coffee with CD36, but not with PPARγ. Except EGCG, the active compound of tea and coffee fulfil the criteria of drug-likeness. The online prediction function demonstrated that secondary metabolites of coffee and tea potentially inhibit foam cell atherosclerosis. No cytotoxicity effect was obtained after the 24 hours treatment of serial dose of DGCTE 10-640 μg/ml (p=0.000). Moreover, the DGCTE (320/320 μg/ml) had a synergistic effect (Loewe score = 17.26417) and significantly reduced the foam cell number in oxLDL-stimulated Raw264.7 compared to control (p=0.000). Conclusion: The DCGTE exhibit potential benefit as candidate agent for the prevention of atherosclerosis-based diseases

ANALISIS DAN KLASIFIKASI SEDIMEN PERMUKAAN DASAR LAUT MENGGUNAKAN SUB-BOTTOM PROFILER

Peningkatan aktivitas manusia di bidang kelautan seperti operasi pengerukan (dredging), eksplorasi minyak dan gas, penambangan pasir mineral, dan berbagai penelitian kelautan telah mengakibatkan permintaan yang mendesak terhadap peta dasar laut yang akurat. Sub-bottom Profiler (SBP) adalah sistem akustik tradisional yang digunakan untuk menggambarkan lapisan sedimen dan batuan di bawah dasar laut, serta memberikan informasi tentang ketebalan sedimen dan stratigrafinya. Oleh karena itu, tujuan dari penelitian ini yaitu menentukan profil lapisan sedimen dasar laut (near-surface sediment) dan memperoleh nilai koefisien refleksi dari lapisan tersebut. Proses akuisisi data dilakukan menggunakan SBP jenis sparker single channel oleh BPPT, yaitu survei pemeliharaan pipa migas dan kabel listrik Pertaminatahun 2010 yang berada di sebelah timur pantai Balongan, Indramayu. Pengambilan sampel sedimen dasar laut menggunakan gravity corer. Hasil dari pengolahan data mentah SBP dengan Matlab di dekat lokasi B19 diperoleh profil 2D dari lapisan sedimen dimana terlihat jelas dasar laut dan reflektornya serta memiliki nilai koefisien refleksi berkisar 0 hingga 0.2955. Selain itu teknik analitis untuk estimasi parameter sedimen dari data laboratorium ditunjukkan sebagai pembanding. Hasil sedimennya berupa silt/lanau dengan nilai koefisien refleksinya adalah 0.2807. </p

Biochemical properties of crude extracellular proteases from Chromohalobacter salexigens BKL5 and Micrococcus luteus 11A

In this work, we have reported an enzymatic activity and biochemical properties of extracellular proteases from Chromohalobacter salexigens BKL5 and Micrococcus luteus 11A. C. salexigens BKL5 and M. luteus 11A were previously isolated from Bledug Kuwu mud volcano and dietary industry wastewater treatment, respectively. Both bacterial strains were able to produce extracellular proteases, when grown on minimal agar medium supplemented with 1% of skim milk. Proteolytic indexes of C. salexigens BKL5 and M. luteus 11A were 2.5±0.14 and 2.9±0.42, respectively. Both extracellular proteases exhibited optimum enzymatic activity at pH 7, with specific activity of C. salexigens BKL5 was 13.3% higher than that of M. luteus 11A. Optimum temperature for enzymatic activity of both proteases was 45°C. Metal cofactor preferences assay showed that extracellular protease from C. salexigens BKL5 preferred Zn2+, meanwhile extracellular protease from M. luteus 11A mainly preferred Ca2+ ion. Metal cofactor preferences assay also suggested that crude extracellular protease from C. salexigens BKL5 was categorized as metalloprotease, meanwhile crude extracellular protease of M. luteus 11A was common neutral protease. The enzymatic stability assay against various salt concentrations showed that crude extracellular protease from C. salexigens BKL5 was more stable than that of M. luteus 11A