After renal injury, selective damage occurs in the proximal tubules as a result of inhibition of glycolysis. The molecular mechanism of damage is not known. Poly(ADP-ribose) polymerase (PARP) activation plays a critical role of proximal tubular cell death in several renal disorders. Here, we studied the role of PARP on glycolytic flux in pig kidney proximal tubule epithelial LLC-PK1 cells using XFp extracellular flux analysis. Poly(ADP-ribosyl)ation by PARP activation was increased approximately 2-fold by incubation of the cells in 10 mM glucose for 30 minutes, but treatment with the PARP inhibitor 3-aminobenzamide (3-AB) does-dependently prevented the glucose-induced PARP activation (approximately 14.4% decrease in 0.1 mM 3-AB-treated group and 36.7% decrease in 1 mM 3-AB-treated group). Treatment with 1 mM 3-AB significantly enhanced the glucose-mediated increase in the extracellular acidification rate (61.1±4.3 mpH/min vs. 126.8±6.2 mpH/min or approximately 2-fold) compared with treatment with vehicle, indicating that PARP inhibition increases only glycolytic activity during glycolytic flux including basal glycolysis, glycolytic activity, and glycolytic capacity in kidney proximal tubule epithelial cells. Glucose increased the activities of glycolytic enzymes including hexokinase, phosphoglucose isomerase, phosphofructokinase-1, glyceraldehyde-3-phosphate dehydrogenase, enolase, and pyruvate kinase in LLC-PK1 cells. Furthermore, PARP inhibition selectively augmented the activities of hexokinase (approximately 1.4-fold over vehicle group), phosphofructokinase-1 (approximately 1.6-fold over vehicle group), and glyceraldehyde-3-phosphate dehydrogenase (approximately 2.2-fold over vehicle group). In conclusion, these data suggest that PARP activation may regulate glycolytic activity via poly(ADP-ribosyl)ation of hexokinase, phosphofructokinase-1, and glyceraldehyde-3-phosphate dehydrogenase in kidney proximal tubule epithelial cells.
Animals ; Cell Death ; Epithelial Cells* ; Glucose ; Glucose-6-Phosphate Isomerase ; Glycolysis ; Hexokinase ; Kidney* ; LLC-PK1 Cells ; Oxidoreductases ; Phosphofructokinase-1 ; Phosphopyruvate Hydratase ; Poly Adenosine Diphosphate Ribose* ; Poly(ADP-ribose) Polymerases* ; Pyruvate Kinase ; Swine

Background: Rotavirus is the main cause of acute viral gastroenteritis in children under 5 years old. The virus leads to over 600000 children deaths a year in the world, 80% of which occur in the developing countries. In Viet Nam, 50%-70% the children\u2019s hospitalizations for acute diarrhea were resulted from rotavirus infection. Objective: To sequence nucleotides and amino acids of VP4, VP7, NSP1, and NSP4 genes of 5 passages of human rotavirus strain G1P8. Materials and method: A study was conducted in rotavirus sample of 5 passages of human rotavirus strain G1P8: B17A3; B17.3; B17.3 pp32vero15; B17.3 pp36TKP2; B17.3 pp43.7vero in Centre for Disease Control and Prevention, Atlanta, United State. Methods: using NucliSen Kit for detection of ARN; RT-PCR; sequencing genes by ABI 3100 machine. Results and Conclusion: Sequencing nucleotides and amino acids of VP4, VP7, NSP1, and NSP4 genes of 5 passages of human rotavirus strain G1P8 showed that: the number of nucleotide mutations ofVP4, VP7, NSP4 genes occurring among the passages were 3 (at nucleotit 175, 419, 790), 1 (at nucleotit 644), 3 (at nucleotit 134, 254, 482), respectively. All these mutations resulted in changes in amino acid composition. No mutation was found in NSP1 gene.
Rotavirus ; Genes ; Nucleotides/ genetics ;

Nucleotide analysis of restricted fragment DNA extremities at definite cut sizes identified: there were 3 extremities: flat; cross with DNA 5’; cross with DNA 5’. This study presented a method which able to characterize unknown restriction enzymes by analyzing the cutting extremities of restricted fragments. The method is based on Sanger sequencing of DNA around the cut site and non-radioactive chemical. Klenow reaction had two active characters: analyzing DNA 5’3’ and exonucleasa 3’5’, allowing to determine the type of cutting of restrictive enzymes. This results allowed to identify the cut sizes of two new enzymes: for Sml1, the cut sites occurs within the recognition nucleotide sequence; BciVI recognizing a non-palindrome sequence and cuts outside this sequence.
DNA ; Nucleotides ; analysis ; extremities ; Enzymes

The morbidity of neurodegenerative diseases are increased in recent years, however, the treatment is limited. Poly ADP-ribosylation (PARylation) is a post-translational modification of protein that catalyzed by poly(ADP-ribose) polymerase (PARP). Studies have shown that PARylation is involved in many neurodegenerative diseases such as stroke, Parkinson's diseases, Alzheimer's disease, amyotrophic lateral sclerosis and so on, by affecting intracellular translocation of protein molecules, protein aggregation, protein activity, and cell death. PARP inhibitors have showed neuroprotective efficacy for neurodegenerative diseases in pre-clinical studies and phase Ⅰ clinical trials. To find new PARP inhibitors with more specific effects and specific pharmacokinetic characteristics will be the new direction for the treatment of neurodegenerative diseases. This paper reviews the recent progress on PARylation in neurodegenerative diseases.
ADP-Ribosylation ; Humans ; Neurodegenerative Diseases ; physiopathology ; Poly Adenosine Diphosphate Ribose ; Poly(ADP-ribose) Polymerases ; metabolism

Measurement of thiopurine metabolites is helpful to monitor adverse effects and assess compliance in patients on thiopurine treatment. The purpose of this study was to develop and validate an analytical method for measurement of thiopurine metabolites, thioguanine nucleotides (6-TGN) and 6-methylmercaptopurine nucleotide (6-MMPN), in RBCs. We developed and validated a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the quantification of 6-TGN and 6-MMPN and evaluated the stability of the thiopurine metabolites in RBC and whole blood states without any preprocessing at various storage conditions. The linear range was 0.1–10 µmol/L and 0.5–100 µmol/L for 6-TGN and 6-MMPN, respectively. The mean extraction recovery at the two concentrations was 71.0% and 75.0% for 6-TGN, and 102.2% and 96.4% for 6-MMPN. Thiopurine metabolites in preprocessed RBC samples were stable at 25℃ and 4℃ after storage for 4 hours and at −70℃ for up to 6 months. However, 6-TGN decreased by 30% compared with the initial concentration when stored at −20℃ for 180 days. In whole blood states, 6-TGN decreased by about 20% at four days after storage at 4℃. We validated a reliable LC-MS/MS method and recommend that the patient's whole blood sample be preprocessed as soon as possible.
Compliance ; Humans ; Mass Spectrometry ; Methods* ; Nucleotides ; Thioguanine

We cultured the rabbit inner medullary collecting duct (IMCD) cells as monolayers on collagen-coated membrane filters, and investigated distribution of the P2Y receptors by analyzing nucleotide-induced short circuit current (Isc) responses. Exposure to different nucleotides of either the apical or basolateral surface of cell monolayers stimulated Isc. Dose-response relationship and cross-desensitization studies suggested that at least 3 distinct P2Y receptors are expressed asymmetrically on the apical and basolateral membranes. A P2Y2-like receptor, which responds to UTP and ATP, is expressed on both the apical and basolateral membranes. In addition, a uracil nucleotide receptor, which responds to UDP and UTP, but not ATP, is expressed predominantly on the apical membrane. In contrast, a P2Y1-like receptor, which responds to ADP and 2-methylthio-ATP, is expressed predominantly on the basolateral membrane. These nucleotides stimulated intracellular cAMP production with an asymmetrical profile, which was comparable to that in the stimulation of Isc. Our results suggest that the adenine and uracil nucleotides can interact with different P2Y nucleotide receptors that are expressed asymmetrically on the apical and basolateral membranes of the rabbit IMCD cells, and that both cAMP- and Ca2+-dependent signaling mechanisms underlie the stimulation of Isc.
Adenine ; Adenosine Diphosphate ; Adenosine Triphosphate ; Membranes ; Nucleotides ; Uracil ; Uracil Nucleotides ; Uridine Diphosphate ; Uridine Triphosphate

CpG is the pair of nucleotides C and G, appearing successively, in this order, along one DNA strand. It is known that due to biochemical considerations CpG is relatively rare in most DNA sequences. However, in particular subsequences, which are a few hundred to a few thousand nucleotides long, the couple CpG is more frequent. These subsequences, called CpG islands, are known to appear in biologically more significant parts of the genome. The ability to identify CpG islands along a chromosome will therefore help us spot its more significant regions of interest, such as the promoters or 'start' regions of many genes. In this respect, I developed the CpG islands search tool, CpG Islands Detector, which was implemented in JAVA to be run on any platform. The window-based graphical user interface of CpG Islands Detector may facilitate the end user to employ this tool to pinpoint CpG islands in a genomic DNA sequence. In addition, this tool can be used to highlight potential genes in genomic sequences since CpG islands are very often found in the 5' regions of vertebrate genes.
Base Sequence ; CpG Islands ; DNA ; Genome ; Indonesia ; Nucleotides ; Vertebrates

A series of modified oligonucleotides containing P=S backbone and a six-membered azasugar (6-AZS) were synthesized and tested for their ability to inhibit human immunodeficiency virus (HIV) in vitro without the aid of any transfecting agents. While P=S oligonucleotides with natural nucleotides had little anti-HIV-1 activity, six-membered azasugar nucleotide (6-AZN)-containing P=S oligonucleotides (AZPSON) potently inhibited the HIV-1/SHIV production and syncytium formation in vitro (EC50 = 0.02~0.2 micro M) without cytotoxicity up to 100 micro M. AZPSONs are enzymatically stable over 6 days in culture supernatant. Phosphodiester (P=O) backbone only or mixed backbone (P=O and P=S) oligonucleotides that contain 6-AZN did not exhibit anti-HIV-1 activity. The anti-HIV-1 capacity of AZPSON seems to depend on the number and/or distribution patterns of 6-AZN in the oligonucleotides. The oligomer 2198, most effective for anti-HIV-1 activity among the AZPSONs, was much more effective than ddI or ddC in anti-HIV activity. Particularly noteworthy is that the anti-HIV-1 activity of AZPSON-2198 was better than AZT in the long-lasting efficacy after a single treatment.
Giant Cells ; HIV ; HIV-1 ; Nucleotides ; Oligonucleotides ; Phosphorothioate Oligonucleotides*

OBJECTIVETo observe therapeutic effect of thread embedding therapy on chronic gastritis.

METHODSSeventy cases of chronic gastritis were randomly divided into the treatment group (n = 36) treated by thread embedding therapy and the control group (n = 34) by acupuncture. Weishu (BL 21), Zhongwan (CV 12) and Zusanli (ST 36) were selected as main points. And plasma contents of cAMP, cGMP, gastrin and substance P were observed before and after treatment.

RESULTSThe total effective rate was 88.89% in the treatment group and 76.47% in the control group with a significant difference between the two groups (P < 0.05); there were significant differences before and after treatment in plasma contents of cAMP, cGMP, gastrin and substance P in the two groups (P < 0.01), and the changes of these indexes in the treatment group were significantly superior to the control group (P < 0.05).

CONCLUSIONThread embedding therapy has a definite therapeutic effect on chronic gastritis and it can adjust nucleotides, gastrin and substance P to improve the functions of the nerve-endocrine-immunity network.

Cultural characteristics and phylogenic relationships were investigated and classified among collected strains in Pholiota spp. which contain P. adiposa, P. squarrosa, P. nameko etc. They were tested on the four different media (PDA, MCM, YM, MEA) and sawdust (Alder, Oak, Pine, Popular) substrates. There was a little variation according to the media and sawdust substrates, although PDA and popular sawdust substrate seemed to be better. Most strains showed white colonies, but some strains were brown. Mycelial growth length differed according to the strains. To classify species, the internal transcribed spacer regions (ITS) of the ribosomal DNA (rDNA) repeats from Pholiota spp. were amplified using polymerase chain reaction (PCR) and then sequenced. According to the analysis of ITS sequences, they were classified into five clusters. Their spacer regions were 644~700 nucleotides in length. The reciprocal homologies of each ITS region among these strains were ranged from 49.6~99.9%. The phylogenic analysis might give a criterion to classify species in the collected strains.
Cultural Characteristics* ; DNA, Ribosomal ; Nucleotides ; Pholiota* ; Polymerase Chain Reaction