1. Rapid Method for the Determination of Cyclophosphamide and Ifosphamide in Urine at Trace Levels by Gas Chromatography-Mass Spectrometry Noushin Rastkari, Reza Ahmadkhaniha, Maryam Alehashem, Shadi Baniasadi
Introduction: The oxazaphosphorines cyclophosphamide (CY) and Ifosfamide (IF) are amongst the most widely used drugs in cancer chemotherapy. In both routine cancer therapy and experimental cancer research, many people might be exposed to CY and IF during pharmaceutical handling of the agents, so developing of a reliable analysis method for biomonitoring is necessary. Materials and Methods: A rapid and simple method for the simultaneous extraction of CP and IP from human urine has been developed using liquid-liquid extraction. The detection limits of cyclophosphamide and ifosphamide in urine samples were 0.5 and 1 ng ml−1, respectively, with a signal-to-noise ratio of 3:1. The regression correlation coefficients (r2) were over 0.99 in all experiments. Conclusion: This method is sensitive enough to determination of low levels of CP and IF in a range of urine concentrations relevant to performing low exposure assessment.
2. The Role of Oxidant on Dipeptidyl Peptidase-4 Enzyme in Benign Prostatic Hyperplasian Through Computational Study Suhargo Listijani, Sumitro S B, Widodo
Dipeptidyl Peptidase-4 (DPP-4) is an epithelial transmembrane protein of many cell types such as gastrointestinal tract, biliary tract, pancreas, kidney, thymus, uterus, prostate and capillary endothelial cells. DPP-4 also found in soluble form of serum and other body fluid. Soluble DPP-4 has not a cytoplasmic domain and transmembrane domain. In diabetes, hydrogen peroxide as oxidant was involved in soluble DPP-4 generation from transmembrane DPP-4. In Benign Prostatic Hyperplasia (BPH), there were decreasing of antioxidant such as superoxide dismutase, catalase and gluthathione peroxidase, so the antioxidants in those diseases unable to discharge the oxidants and there was an accumulation of oxidants. This study aims to evaluate the role of some oxidant on DPP-4 enzyme in BPH through computational approach. The oxidants such as H2O2, OH–, HOCl, NO–, NO2 ONOO–, 13-hydroperoxy linoleic acid and 13-hydroxy linoleic acid (13-HODE) were docked with dipeptidyl peptidase-4. The result showed that all the oxidants may interact with DPP-4 on hydrolase domain (Gln 508 – Pro 766) or on β-propeller domain (Arg 54 – Asn 497). But 13-HODE may interact with DPP-4 on the stalk of DPP-4 that were on Asp 34, Ala 35, Thr 36, dan Ala 37 residues. So it may cause DPP-4 shedding to produce soluble DPP-4.
3. Computational Analysis For Revealing The Role of Thymoquinone (Active Compound From Ethanolic Extract of Nigella Sativa) As Inhibitor of P65 Nf-Kb Activation in Preclampsia Treatment Indrawan I Wayan Agung, Handono Kalim, Siti Chandra WB, Bambang Rahardjo
Thymoquinone is the main active compound from Nigella sativa and can be used as traditional medice such as preclamsia treatment. Inflammation is one of the pathophysiologic process in Preeclampsia. Preeclampsia is a sistemic inflammatory disease that induces endothelial dysfunction as the main disorder. The aim of this study is to evaluate the role of thymoquinone as antiinflammatory through computational study. The method for predicting the activity is Pass Server online that can predict activity of chemical compound based on structure activity relationship. Model of thymoquine was retrieved from pubchem and model of NF-kB inhibitor was collected from protein data bank. The prediction of protein-protein interaction was conducted using string db and molecular docking analysis for evaluating the potential inhibition of thymoquinone was done using patchdock and firedock web service. Molecular interaction was done using ligplot and the visualization of biomolecules was used PyMol, Chimera and Ligand Scout. The result showed that thymoquinone have antiinflammatory activity indicated by probability score (Pa) 0.6. Docking result showed that thymoquinone can bind to NF-kB inhibitor with high binding affinity (-4.10 Kcal/mol) and it interact with threonin 39 using hydrogen bond and three hydrophobic interaction. The complex can inhibit the phosphorylation process of NF-kB inhibitor. It can be concluded that Thymoquinone is potential for inhibiting the p65 NF-kB activation through inhibit Ikb. Thymoquinine from nigella sativa extract can be a candidate for preclampsia treatment in the future.
4. Periodic Fever, Aphtous Stomatitis, Pharyngitis and Cervical Adenitis – (PFAPA) Syndrome Clinical Cases. Markov S., Dzambazov K., Delev D., Stoyanova-Deleva A., Chatalbashev D
The periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis (PFAPA) syndrome was originally described by Marshall et al in 1987 and the acronym, PFAPA, was coined two years later together with the diagnostic criteria. PFAPA syndrome is regarded as a non-hereditary disease of unknown etiology although in a small proportion of cases, one of the parents or a more distant relative had similar symptoms in childhood. The onset is generally before the age of 5 years, with attacks lasting 3-6 days, recurring every 3-8 weeks. Patients are asymptomatic between episodes. For now the syndrome is considered as a sporadic disease.
5. Secondary Metabolites Analysis of Saccharomyces cerievisiae and Evaluation of Antibacterial Activity Mohammad J. Al-Jassani, Ghaidaa Jihadi Mohammed, Imad Hadi Hameed
The objectives of this study were analysis of the secondary metabolite products and evaluation antibacterial activity. Bioactives are chemical compounds often referred to as secondary metabolites. Twenty one bioactive compounds were identified in the methanolic extract of Saccharomyces cerevisiae. The identification of bioactive chemical compounds is based on the peak area, retention time molecular weight and molecular formula. GC-MS analysis of Saccharomyces cerevisiae revealed the existence of the Thieno [2,3-c] furan-3-carbonitrile, 2-amino-4,6-dihydro-4,4,6,6-, Oxime-, methoxy-phenyl-,Acetic acid, N’-[3-(1-hydroxy-1-phenylethyl)phenyl] hydrazide, 1-Aminononadecane, N-trifluoroacetyl, Androstane-11,17-dione,3- [(trimethylsilyl)oxy]-,17-[O-(phenylme, Benzeneacetamide,α-ethyl-, 4-Benzyloxy-6-hydroxymethyl-tetrahydropyran-2,3,5-triol, 1,2-Ethanediol, 1-(2-phenyl-1,3,2-dioxaborolan-4-yl)-,[S-(R*,R*)], Erythritol, 3,6,9,12,-Tetraoxatetradecan-1-ol,14- [4-(1,1,3,3- tetramethylbutyl , Urea,N,N’-bis(2-hydroxyethyl)- , Ergosta-5,22-dien-3-ol,acetate,(3β,22E)-, Ethyl iso-allocholate, (5β)Pregnane-3,20β-diol,14α,18α-[4-methyl-3-oxo-(1-oxa-4-azal,5,5′-Dimethoxy-3,3′,7,7′-tetramethyl-2,2′-binaphthalene-1,1′,4,4′,N-(4,6-Dimethyl2pyrimidinyl)-4-(4 nitrobenzylideneamino) benzene, 3-[3-Bromophenyl]-7-chloro-3,4-dihydro-10-hydroxy-1,9 (2H,10H) , 2-Methyl-9-β-d-ribofuranosylhypoxanthine, Dodecane,1-chloro-, 2,7-Diphenyl-1,6-dioxopyridazino [4,5:2′,3′] pyrrolo [4′,5′-d] pyridazin and 2-Bromotetradecanoic acid. Evernia punastri was very highly antifungal activity (7.00±0.25) mm. The results of anti-bacterial activity produced by Saccharomyces cerevisiae showed that the volatile compounds were highly effective to suppress the growth of Proteus mirabilis.
6. In Vitro Study of NF-κB and Pro-Inflammatory Cytokines Declining Levels by Polyherbal Emsa Eritin Muhaimin Rifa’I, Yuyun Ika Christina, Qonitatul Khasanah, Agung Pramana Warih Marhendra, Hideo Tsuboi
Inflammation is an immunological response which plays pivotal role in antigen elimination, but it also causes detrimental effect to the surrounding tissue. Inflammation pathway is activated by the molecular signaling which regulate both pro- and anti-inflammatory cytokines. NF-κB is a transcription factor which regulates the transcription of several genes that regulate growth, angiogenesis, and survival, including the transcription of inflammatory cytokines. In the present study, we examined the effect of polyherbal EMSA Eritin focusing on immunological function with aim of highlighting the production of pro-inflammatory cytokines and the expression of transcriptional factor NF-κB in vitro. EMSA Eritin is a polyherbal consisted of soybean, red rice, and coconut water extract. Splenocytes from healthy mice were cultured for 5 days in RPMI-1640 medium. On day 5, the cultured cells were harvested and analyzed by flow cytometry. EMSA Eritin was able to inhibit the expression of NF-κB in T cells and also the production of proinflammatory cytokines marked by the decreasing levels of CD4+IFN-γ+ and CD4+TNF-α+ T cells. Previous study indicated that EMSA Eritin is able to decrease NF-κB and pro-inflammatory cytokines in irradiated mice (in vivo study) with sublethal dose. Hereby, our study suggests that EMSA Eritin is a potential herbal medicine which act as an anti-inflammatory agent.
7. Comparative Study of the Results of Capillary Blood International Normalized Ratio on Coagucheck XS POC and Conventional Coagulation Blood Test Dinkova A, Vladimirova-Kitova L, Deneva-Koycheva T
Millions of patients worldwide are taking anticoagulants like acenocumarol and warfarin for prevention of serious thrombotic complications as stroke or heart attack. The coumarins or vitamin K antagonists (VKAs) have been the mainstay of oral anticoagulant therapy for more than 50 years. In clinical practice VKAs are challenging to use, because they reveal considerable variability in dose response among different patients, resulting from genetic mutation and drug and diet interactions. Because of this they require frequent laboratory monitoring which is time consuming and discomfort for the patients. The development of point-of-care (POC) devices enables International normalized ratio (INR) to be obtained in outpatient as well as at home setting at the moment. Some earlier studies have shown statistically significant differences between values from clinical laboratory and POC meters, because of which the accuracy of these meters remain still controversial. The aim of this study was to evaluate the results of INR on the CoaguChek XS meter relative to chronometric assay on laboratory coagulometer Sysmex S 2000i.