Objektif kajian ialah untuk menentukan kesan program latihan papan Wobble ke atas daya keseimbangan para atlet yang mengalami ketakstabilan fungsi pergelangan kaki (FAI). Sebanyak 30 subjek (n=30) yang mengalami FAI direkrut berdasarkan kriteria rangkuman dan eksklusi. Kesemua subjek dinilai daya keseimbangan menggunakan ujian dirian anggota tunggal (single limb stance) dan ujian jangkauan fungsi dan didedahkan kepada program latihan papan Wobble sebanyak lima sesi seminggu selama empat minggu. Ujian t berpasangan digunakan untuk membandingkan min data pra dan post-ujian dan aras signifikan ditetapkan pada nilai 0.05. Pada ujian dirian anggota tunggal, para atlet mendapat min skor postlatihan yang tinggi [26.4] berbanding pra-latihan. Perbezaan disahkan dengan nilai ‘t’ yang diperolehi [16.00]. Bagi ujian jangkauan fungsi, para atlet memperolehi min skor post-latihan yang tinggi [15.88] berbanding pra-latihan [1.26]. Perbezaan disahkan dengan nilai ‘t’ yang diperolehi [12.54]. Ini menunjukkan bahawa latihan papan Wobble meningkatkan keseimbangan dinamik dan statik para atlet yang mengalami FAI. Secara kesimpulannya, latihan papan Wobble mungkin efektif dalam meningkatkan keseimbangan para atlet yang mengalami FAI.
Telbivudine, a thymidine nucleoside analog, is a common therapeutic option for chronic hepatitis B infection. While raised serum creatine kinase is common, myopathy associated with telbivudine is rare. Reports on its myopathological features are few and immunohistochemical analyses of inflammatory cell infiltrates have not been previously described. We describe the clinical, myopathological and immunohistochemical features of four patients who developed myopathy after telbivudine therapy for chronic hepatitis B infection. All four patients presented with progressive proximal muscle weakness, elevation of serum creatine kinase and myopathic changes on electromyography. Muscle biopsies showed myofiber degeneration/necrosis, regeneration, and fibers with cytoplasmic bodies and cytochrome c oxidase deficiency. There was minimal inflammation associated with strong sarcolemmal overexpression of class I major histocompatibility complex (MHC class I). Upon withdrawal of telbivudine, muscle weakness improved in all patients and eventually completely resolved in three. In our series, telbivudine-associated myopathy is characterized by necrotizing myopathy which improved on drug withdrawal. Although the occasional loss of cytochrome c oxidase is consistent with mitochondrial toxicity, the overexpression of MHC class I in all patients could suggest an underlying immune-mediated mechanism which may warrant further investigation.
An in-house method which utilizes 14C-thymidine as a substrate was used to assay deoxythymidine kinase in serum. The method is sensitive enough to detect normal levels of serum deoxythymidine kinase and the assay procedure also enables rapid handling of multiple samples. With a total reaction volume of 60 ul, the enzyme reaction was found to be linear with concentrations for up to 650 U/L of TK activity. On studying serum deoxythymidine kinase (s-TK) activity with incubation time, there was a proportional increase in activity with the length of incubation time. "Within-batch" precision showed a coefficient of variation (CV) of 4.7% for serum with extremely high s-TK levels and a CV of 8.8% for serum with normal s-TK levels. S-TK showed a CV of less than 16.0% in its activity when stored at -8 degrees C and at -20 degrees C. The normal reference range obtained for s-TK activity was 8.6 +/- 7.5 U/L.
In vitro lymphocyte proliferative response of peripheral blood leucocytes (PBL) to purified Toxoplasma gondii antigen were evaluated by 3[H] methyl thymidine incorporation in patients acutely and chronically infected with Toxoplama gondii. PBL from three patients with acute sylnptonlatic toxoplaslnosis showed no response to T. gondii antigen during the emergence of anti-Toxoplasma 1gM antibodies and the response returned as the infection became chronic. Lymphocytes of twelve chronically-infected patients responded positively to the antigen. In all patients the lymphocyte proliferative response to the mitogen, Concanavalill A (Con A) was normal. Analysis of Toxoplasma proliferative response of PBL from a patient with acute toxoplasrnosis showed that CD8+ cells were responsible for induction of suppression while the response during the chronic infection was lnediated by CD4+ cells. In human toxoplasmosis there was antigen-specific lymphocyte unresponsiveness during the acute phase of the infection and it appears that the initnunesuppression was mediated by CD8+ cells. KEYWORDS: Toxoplasmosis-lymphocyte blastogenesis-antigen specific-CD4+, CD8+
Thymidine phosphorylase (TP) over expression plays role in several pathological conditions, such as rheumatoid arthritis, chronic inflammatory diseases, psoriasis, and tumor angiogenesis. The inhibitor of this enzyme plays an important role in preventing the serious threat due to over expression of TP. In this regard, a series of seventeenanalogs of 3-formylcoumarin (1-17) were synthesized, characterized by 1HNMR and EI-MS and screened for thymidine phosphorylaseinhibitory activity. All analogs showed a variable degree of thymidine phosphorylase inhibition with IC50 values ranging between 0.90 ± 0.01 and 53.50 ± 1.20 μM when compared with the standard inhibitor 7-Deazaxanthine having IC50 value 38.68 ± 1.12 μM. Among the series, fifteenanalogs such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 16 and 17 showed excellent inhibition which is many folds better than the standard 7-Deazaxanthine whiletwo analogs 13 and 14 showed good inhibition. The structure activity relationship (SAR) was mainly based upon by bring about difference of substituents on phenyl ring. Molecular docking study was carried out to understand the binding interaction of the most active analogs.
Lichens are symbiotic organisms that have been traditionally used for treating different kinds of ailments. As there are only a few reports on the antiviral activity of lichens, we thought of evaluating the anti-Herpes simplex virus-1 (HSV-1) activity of methanolic extract of Roccella montagnei and their isolated compounds. Fractionation of crude methanolic extract of Roccella montagnei by column chromatography isolated two pure compounds. Antiviral activity was assessed using a CPE inhibition assay at non-cytotoxic concentrations on Vero cells. Molecular docking and dynamics studies were carried out against Herpes simplex type-1 thymidine kinase to understand the binding interactions of the isolated compounds with reference to acyclovir. Isolated compounds were characterized as methyl orsellinate and montagnetol by spectral methods. Methanolic extract of Roccella montagnei exhibited an EC50 value of 56.51 µg/ml, while the compounds methyl orsellinate and montagnetol offered EC50 values of 13.50 µg/ml and 37.52 µg/ml, respectively, against HSV-1 viral infection on Vero cell lines. The selectively index (SI) of montagnetol (10.93) was found to be higher when compared to that of methyl orsellinate (5.55), indicating its better anti-HSV-1 activity. The docking and dynamics studies showed montagnetol was stable throughout the 100 ns, having better interactions and docking scores with HSV-1 thymidine kinase than methyl orsellinate, as well as the standard. To understand the mechanism of montagnetol's anti-HSV-1 activity, more research is required, and this could lead to the discovery of new and effective antiviral agents.Communicated by Ramaswamy H. Sarma.
Matched MeSH terms: Thymidine Kinase/pharmacology; Thymidine Kinase/therapeutic use
In this study, we investigated the potential in vitro anti-HSV-1 activities of the Cassiopea andromeda jellyfish tentacle extract (TE) and its fractions, as well as computational work on the thymidine kinase (TK) inhibitory activity of the identified secondary metabolites. The LD50, secondary metabolite identification, preparative and analytical chromatography, and in silico TK assessment were performed using the Spearman-Karber, GC-MS, silica gel column chromatography, RP-HPLC, LC-MS, and docking methods, respectively. The antiviral activity of TE and the two purified compounds Ca2 and Ca7 against HSV-1 in Vero cells was evaluated by MTT and RT-PCR assays. The LD50 (IV, mouse) values of TE, Ca2, and Ca7 were 104.0 ± 4, 5120 ± 14, and 197.0 ± 7 (μg/kg), respectively. They exhibited extremely effective antiviral activity against HSV-1. The CC50 and MNTD of TE, Ca2, and Ca7 were (125, 62.5), (25, 12.5), and (50, 3.125) μg/ml, respectively. GC-MS analysis of the tentacle extract revealed seven structurally distinct chemical compositions. Four of the seven compounds had a steroid structure. According to the docking results, all compounds showed binding affinity to the active sites of both thymidine kinase chains. Among them, the steroid compound Pregn-5-ene-3,11-dione, 17,20:20,21 bis [methylenebis(oxy)]-, cyclic 3-(1,2-ethane diyl acetal) (Ca2) exhibited the highest affinity for both enzyme chains, surpassing that of standard acyclovir. In silico data confirmed the experimental results. We conclude that the oxosteroid Ca2 may act as a potent agent against HSV-1.
Thymidine phosphorylase (TP) is up regulated in wide variety of solid tumors and therefore presents a remarkable target for drug discovery in cancer. A novel class of extremely potent TPase inhibitors based on benzopyrazine (1-28) has been developed and evaluated against thymidine phosphorylase enzyme. Out of these twenty-eight analogs eleven (11) compounds 1, 4, 14, 15, 16, 17, 18, 19, 20, 24 and 28 showed potent thymidine phosphorylase inhibitory potentials with IC50 values ranged between 3.20±0.30 and 37.60±1.15μM when compared with the standard 7-Deazaxanthine (IC50=38.68±4.42μM). Structure-activity relationship was established and molecular docking studies were performed to determine the binding interactions of these newly synthesized compounds. Current studies have revealed that these compounds established stronger hydrogen bonding networks with active site residues as compare to the standard compound 7DX.
Inhibition of Thymidine phosphorylase (TP) is continuously studied for the design and development of new drugs for the treatment of neoplastic diseases. As a part of our effort to identify TP inhibitors, we performed a structure-based virtual screening (SBVS) of our compound collection. Based on the insights gained from structures of virtual screening hits, a scaffold was designed using 1,3,4-oxadiazole as the basic structural feature and SAR studies were carried out for the optimization of this scaffold. Twenty-five novel bis-indole linked 1,3,4-oxadiazoles (7-31) were designed, synthesized and tested in vitro against E. coli TP (EcTP). Compound 7 emerged as potent TP inhibitor with an IC50 value of 3.50 ± 0.01 μM. Docking studies were carried out using GOLD software on thymidine phosphorylase from human (hTP) and E. coli (EcTP). Various hydrogen bonding, hydrophobic interactions, and π-π stacking were observed between designed molecules and the active site amino acid residues of the studied enzymes.
We have synthesized oxadiazole derivatives (1-16), characterized by 1H NMR, 13C NMR and HREI-MS and screened for thymidine phosphorylase inhibitory potential. All derivatives display varied degree of thymidine phosphorylase inhibition in the range of 1.10 ± 0.05 to 49.60 ± 1.30 μM when compared with the standard inhibitor 7-Deazaxanthine having an IC50 value 38.68 ± 1.12 μM. Structure activity relationships (SAR) has been established for all compounds to explore the role of substitution and nature of functional group attached to the phenyl ring which applies imperious effect on thymidine phosphorylase activity. Molecular docking study was performed to understand the binding interaction of the most active derivatives with enzyme active site.
Thymidine phosphorylase (TP, EC 2.4.2.4), an enzyme involved in pyrimidine salvage pathway, is identical to platelet-derived endothelial cell growth factor (PD-ECGF) and gliostatin. It is extremely upregulated in a variety of solid tumours. The TP amplification is associated with concomitant overexpression of many angiogenic factors such as matrix metalloproteases (MMPs), interleukins (ILs), vascular endothelial growth factor (VEGF) etc., resulting in promotion of angiogenesis and cancer metastasis. In addition, overshooting TP level protects tumour cells from apoptosis and helps cell survival. Thus, TP is identified as a prime target for developing novel anticancer therapies. Pioneering research activities investigated a large number of TP inhibitors, most of which are pyrimidine or purine analogues. Recently, an array of structurally diverse non-nucleobase derivatives was designed, synthesized and established as promising TP inhibitors. This review, following an outline on the TP structure and functions, gives an overview of the recent advancement of various non-nucleobase TP inhibitors as novel anti-cancer agents.
Metaldehyde is used widely in Malaysia for the control of molluscs. This communication reports the cytotoxic effects of this chemical on cultured cells as assessed by cell morphology and the DNA synthesising capability as well as its transport into cells. After 15 days of exposure with 20.0 ppm of the compound, the DNA synthesising capability was shown to be unaffected. The IC50 for Vero cells was 276.0 ppm. Transport of thymidine across cells was found to be not significantly affected even at high metaldehyde concentrations (up to 320.0 ppm) suggesting integrity of cells were not significantly affected. The present cellular studies have therefore shown that the cytotoxic effects of this chemical is rather low.
Metaldehida digunakan dengan meluas di Malaysia untuk mengawal perosak moluska. Kesan sitotoksik bahan kimia ini di peringkat sel dari segi ciri-ciri perubahan moifologi dan keupayaan mensintesis DNA serta kajian awal kesannya terhadap proses kemasukan ke dalam sel dilaporkan di sini. Keupayaan mensintesis DNA didapati tidak terjejas secara signifikan selepas diberikan 20.0 ppm metaldehida secara berterusan selama 15 hari. Nilai IC50 bagi sel Vero adalah 276.0 ppm. Kemasukan timidina ke dalam sel tidak terjejas secara signifikan apabila sel diperlakukan dengan metaldehida, walaupun pada kepekatan yang agak tinggi iaitu sehingga 320.0 ppm. Kajian telah menunjukkan bahawa kesan sitotoksik oleh metaldehida adalah rendah.
5'-Phosphodiesterase (5'-PDE) is an enzyme that hydrolyses RNA to form 5'-inosine monophosphate (5'-IMP) and 5'-guanosine monophosphate (5'-GMP), which function as flavour enhancers. Selection of the best producer of 5'-PDE was made by determining the activity of the enzyme in six seeds that have been germinated, namely mung bean (Vigna radiate), soybean (Glycine max), adzuki/red bean (Vigna angularis L.), chick pea (Cicer arietinum), black eye pea (Vigna unguiculata) and petai (Parkia speciosa). Seeds that were not germinated acted as the control. In order to ensure there is no contamination from potential 5'-PDE-producing microorganisms during germination, microbial growth was reduced by using different surface sterilizing treatments where the seeds were soaked in 100 mL solution containing different concentrations of sodium hypochlorite (with or without 0.05% sodium azide) for 5 minutes before rinsing it five times with sterilized distilled water (total 500 mL). The seeds were observed every day for 3 days and the best surface sterilizing treatment was selected based on absence of mold growth and the effects on hypocotyl length. Sodium hypochlorite at 0.3% (v/v) concentration was able to inhibit mold growth in adzuki bean, soybean and chickpea. On the other hand, only 0.1% (v/v) sodium hypochlorite was needed to inhibit mold growth in black eye pea and petai, while mung bean required 0.05% (v/v) sodium hypochlorite to inhibit mold growth. Under these conditions, the growth of hypocotyl (hypocotyls length) was only slightly affected compared to the control. 5'-PDE was extracted from seeds that have been germinated for 24 hours and their control (ungerminated seeds) by homogenization in a blender with 400 mL of 50 mM acetate buffer, pH 4.5. After that, the homogenates were stirred for 30 min and the centrifuged at 9000 rpm for 15 min at 10°C. 5'-PDE activity was determined using thymidine 5'-monophosphate p-nitrophenyl ester as substrate at pH 7.0 and 55°C. The formation of nucleotide monophosphates, the products of reaction, was determined at 405 nm. As a strong presence of phosphomonoesterase (PME) will reduce the yield of nucleotide monophosphates as the enzyme hydrolyzes these products into nucleosides and orthophosphate, PME activity was also determined using p-nitrophenyl phosphate as the substrate at 60°C and pH 5.0. Thus, the seed with the highest 5'-PDE activity and a low PME activity can be selected. Germinated adzuki bean was found to have the highest 5'-PDE activity (0.59 µmol p-nitrophenol/min/mg protein) among the germinated seeds. A time-course study indicated that the level of 5'-PDE in adzuki bean increased with time of germination until 15 hours (0.69 µmol p-nitrophenol/min/mg protein), after which the acitivity decreased until it reached the basal level (0.44 µmol p-nitrophenol/min/mg protein) at 72 hours. On the other hand, PME in the bean was the highest at 9 h germination (0.98 µmol p-nitrophenol/min/mg protein). In general, controls have very low basal level of 5'-PDE activity (0.18- 0.42 µmol p-nitrophenol/min/mg protein).
Conventional therapies for malignant cancer such as chemotherapy and radiotherapy are associated with poor survival rates owing to the development of cellular resistance to cancer drugs and the lack of targetability, resulting in unwanted adverse effects on healthy cells and necessitating the lowering of therapeutic dose with consequential lower efficacy of the treatment. Gene therapy employing different types of viral and non-viral carriers to transport gene(s) of interest and facilitating production of the desirable therapeutic protein(s) has tremendous prospects in cancer treatments due to the high-level of specificity in therapeutic action of the expressed protein(s) with diminished off-target effects, although cancer cell-specific delivery of transgene(s) still poses some challenges to be addressed. Depending on the potential therapeutic target genes, cancer gene therapy could be categorized into tumor suppressor gene replacement therapy, immune gene therapy and enzyme- or prodrug-based therapy. This review would shed light on the current progress of delivery of potentially therapeutic genes into various cancer cells in vitro and animal models utilizing a variety of viral and non-viral vectors.
Here in this study regarding the over expression of TP, which causes some physical, mental and socio problems like psoriasis, chronic inflammatory disease, tumor angiogenesis and rheumatoid arthritis etc. By this consideration, the inhibition of this enzyme is vital to secure life from serious threats. In connection with this, we have synthesized twenty derivatives of isoquinoline bearing oxadiazole (1-20), characterized through different spectroscopic techniques such as HREI-MS, 1H- NMR and 13C-NMR and evaluated for thymidine phosphorylase inhibition. All analogues showed outstanding inhibitory potential ranging in between 1.10 ± 0.05 to 54.60 ± 1.50 µM. 7-Deazaxanthine (IC50 = 38.68 ± 1.12 µM) was used as a positive control. Through limited structure activity relationships study, it has been observed that the difference in inhibitory activities of screened analogs are mainly affected by different substitutions on phenyl ring. The effective binding interactions of the most active analogs were confirmed through docking study.
Thymidine phosphorylase is an enzyme involved in pyrimidine salvage pathway that is identical to platelet-derived endothelial cell growth factor (PD-ECGF) and gliostatin. It is enormously up regulated in a variety of solid tumors. Furthermore, surpassing of TP level protects tumor cells from apoptosis and helps cell survival. Thus TP is identified as a prime target for developing novel anticancer therapies. A new class of exceptionally potent isatin based oxadiazole (1-30) has been synthesized and evaluated for thymidine phosphorylase inhibitory potential. All analogs showed potent thymidine phosphorylase inhibition when compared with standard 7-Deazaxanthine, 7DX (IC50 = 38.68 ± 1.12 µM). Molecular docking study was performed in order to determine the binding interaction of these newly synthesized compounds, which revealed that these synthesized compounds established stronger hydrogen bonding network with active site of residues as compare to the standard compound 7DX.
Isoquinoline analogues (KA-1 to 16) have been synthesized and evaluated for their E. coli thymidine phosphorylase inhibitory activity. Except compound 11, all other analogs showed outstanding thymidine inhibitory potential ranging in between 4.40 ± 0.20 to 69.30 ± 1.80 µM when compared with standard drug 7-Deazaxanthine (IC50 = 38.68 ± 4.42 µM). Structure Activity Relationships has been established for all compounds, mainly based on substitution pattern on phenyl ring. All analogs were characterized by various spectroscopic techniques such as 1H NMR, 13C NMR and EI-MS. The binding interactions of isoquinoline analogues with the active site of TP enzyme, the molecular docking studies were performed. Furthermore, the angiogenic inhibitory potentials of isoquinoline analogues (KA-1-9, 14, 12 and 16) were determined in the presence of standard drug Dexamethasone based on percentage inhibitions at various concentrations. Herein this work analogue KA-12, 14 and 16 emerged with most potent angiogenic inhibitory potentials among the synthesized analogues.
Palm oil-based Trimethylolpropane ester (TMP ester), with an iodine value of 66.4 g/100g, was epoxidizedto produce epoxidized TMP esters. In situ epoxidation method was used with peracetic acid to eliminatefatty acid double bonds in palm oil-based TMP ester and change it into oxirane ring. This was done toimprove the oxidative stability of trimethylolpropane ester which is a key concern limiting the usefulservice life in lubricants. The epoxidation was performed by reacting acetic acid as active oxygen carrierwith concentrated hydrogen peroxide as oxygen donor and a small amount of homogeneous catalyst(sulphuric acid). The effects of various parameters on the rate of epoxidation (such as the ratio of moleacetic acid to ethylenic unsaturation, hydrogen peroxide to ethylenic unsaturation and acetic acid moleratio, and amount of catalyst) were studied. The rate of oxidation was investigated by the percentageof oxirane oxygen analysis and iodine value.
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare, autosomal recessive disorder associated with mutations in the thymidine phosphorylase (TYMP) gene. The main objective of this study was to characterize the genetic profiles of the deceased proband's family members (N = 4) using DNA sequencing and to determine miRNA deregulation in MNGIE using miRNA microarray profiling and bioinformatic analysis. We found that the genetic profile of the younger sister showed similar TYMP gene mutations as that of the proband with the exception of a heterozygous mutation in exon 10. The miRNA microarray revealed 55 significantly up-regulated and 65 significantly down-regulated miRNAs. These miRNAs have been implicated in various mitochondrial dynamics such as energy metabolism, Krebs cycle, mitochondria-associated apoptosis, and mitophagy. In conclusion, we demonstrate that blood miRNAs are deregulated in the pathogenesis of MNGIE and these changes may have therapeutic implications. Further experimental studies will be required to elucidate the functional miRNA-mRNA interactions in MNGIE.
1. The effects of alpha-tocopherol and gamma-tocotrienol on glutathione S-transferase (GST) and gamma-glutamyl transpeptidase (gamma-GT) activities in cultured hepatocytes prepared from rats treated with diethylnitrosamine (DEN) and 2-acetylaminofluorene (AAF) were investigated. 2. Both the alpha-tocopherol and gamma-tocotrienol treated hepatocytes showed significantly higher (P < 0.05) GST activities than untreated hepatocytes prepared from the carcinogen treated rats in the first 3 days of culture. Treatment with alpha-tocopherol and gamma-tocotrienol generally resulted in a tendency to increase the GST activities above that in the untreated hepatocytes. 3. Treatment with high doses (125-250 microM) of alpha-tocopherol and low doses (12.5-25 microM) of gamma-tocotrienol generally resulted in a significant reduction in gamma-GT activities at 1-3 days. gamma-GT activities are reduced as the dose of alpha-tocopherol and gamma-tocotrienol are increased.