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Comparison of the mass spectrum from hydrogenated and non-hydrogenated samples showed that the TMS ether of methyl 5,8-dihydroxy octadecanoate was derived from the TMS ether of methyl 5,8-dihydroxy-9,12-octadecadienoate. This was evidenced by the molecular ion at m/z 470 and by the characteristic fragments resulting from cleavage around the double bonds and oxygenated C atoms [8]. Thus RP-HPLC peak 2 (Fig.

1) proved to be 5,8-diHOD. RP-HPLC peak 2* was analyzed as a part of RP-HPLC peak 2, due to overlap. Hydrogenation of the TMS ether derivative showed peaks stemming from cleavage around an oxygenated C-atom. The molecular ion at m/z 370 evidenced that this compound was TMS ether of lactonized 5,8-dihydroxyoctadecanoate. selleck inhibitor Comparing the hydrogenated sample with the non-hydrogenated sample showed that TMS ether of lactonized check details 5,8-dihydroxy octadecanoate probably originated from lactonized 5,8-diHOD. GC/MS analysis of monohydroxy fatty acids (RP-HPLC peak 3) In the GC chromatogram of the hydrogenated monohydroxy fatty acids of RP-HPLC peak 3 (Fig. 1) as TMS ethers of methyl ester derivatives, one prominent peak was present. The mass spectrum identified it as a mixture of the TMS ethers of methyl 8-hydroxy octadecanoate,

methyl 10-hydroxy octadecanoate and a small amount of methyl 9-hydroxy octadecanoate. Also, a small peak of methyl 13-hydroxy octadecanoate was present in the GC chromatogram. In the GC/MS analysis of the corresponding non-hydrogenated monohydroxy fatty acids as TMS ethers of methyl ester derivatives, three peaks were visible in the GC chromatogram. FHPI concentration reference compounds indicated that GC peak 1 (18.3 min) was TMS ether of methyl 8-hydroxy octadecadienoate because of the fragmentation pattern and retention time of the non-hydrogenated sample [7]. The mass spectrum of Tolmetin TMS ether of methyl 10-hydroxy octadecanoate, GC peak 2 (18.4 min), showed that this compound originated from 10-hydroxy octadecadienoic acid (10-HOD). The mass spectrum of GC peak 4

(19.1 min) and the mass spectra of reference compounds showed that TMS ethers of methyl 13-hydroxy octadecanoate and methyl 9-hydroxy decanoate were derived from 13-hydroxy octadecadienoic acid (13-HOD) and 9-hydroxy octadecadienoic acid (9-HOD), respectively. Thus, RP-HPLC peak 3 (Fig. 1) was composed of 8-HOD (20), 10-HOD (18), 13-HOD (1) and 9-HOD (1). GC/MS analysis of monohydroxy fatty acids eluting after RP-HPLC peak 3 (Fig. 1) as TMS ethers of methyl ester derivatives showed that a small amount of 8-HOM was also present (data not shown). Characteristics of oxylipin formation Incubation with [U-13C] 18:2 showed that all oxygenated fatty acid products (RP-HPLC peak 1 to peak 3, Fig. 1) represented a mixture of converted 18:2 from endogenous and exogenous sources. The conversion of 500 nmol exogenously supplied 18:2 was about 50% of the total conversion, as judged by the ratio of [U-13C] labeled fragments to unlabeled fragments on GC/MS.

The potential see more energy of a particle in the spherical coordinates has the following form: (1) where R 0 is the radius of a QD. The radius of a QD and effective Bohr radius of a Ps

a p play the role of the problem parameters, which radically affect the behavior of the particle inside a QD. In our model, the criterion of a Ps formation possibility is the ratio of the Ps effective Bohr radius and QD radius (see Figure 1a). In what follows, we analyze the problem in two SQ regimes: strong and weak. Figure 1 The electron-positron pair in the (a) spherical QD and (b) circular QD. Strong size quantization regime LY2874455 clinical trial In the regime of strong SQ, when the condition R 0 ≪ a p takes place, the energy of the Coulomb interaction between an electron and positron is much less than the energy caused by the SQ contribution. In this approximation, the Coulomb interaction between the electron and positron can be neglected. The problem then

reduces to the determination of an electron and positron energy states separately. As noted above, the dispersion law for narrow-gap semiconductors is nonparabolic and is given in the following form [11, 36]: (2) where S ~ 108 cm/s is the parameter related to the semiconductor bandgap . Let us write the Klein-Gordon equation P505-15 nmr [43] for a spherical QD consisting of InSb with electron and positron when their Coulomb interaction is neglected: (3) where P e(p) is the momentum operator of the particle (electron, positron), is the effective Nintedanib (BIBF 1120) mass of the particle, and E is the total energy of the system. After simple transformations, Equation 3 can be written as the reduced Schrödinger equation: (4) where , is the effective Rydberg energy of a Ps, κ is the dielectric constant

of the semiconductor, and is a Ps effective Bohr radius. The wave function of the problem is sought in the form . After separation of variables, one can obtain the following equation for the electron: (5) where is a dimensionless energy. Seeking the wave function in the form , the following equation for the radial part of (5) could be obtained: (6) Here, , l is the orbital quantum number, m is magnetic quantum number, is the reduced mass of a Ps, is dimensionless bandgap width, is the analogue of fine structure constant, and is the analogue of Compton wavelength in a narrow bandgap semiconductor with Kane’s dispersion law. Solving Equation 6, taking into account the boundary conditions, one can obtain the wave functions: (7) where , J l + 1/2(z) are Bessel functions of half-integer arguments, and Y lm (θ, φ) are spherical functions [44]. The following result could be revealed for the electron eigenvalues: (8) where α n,l are the roots of the Bessel functions.

The isthmal epithelium of the oviduct was washed extensively with HBSS containing 200 U/ml penicillin and 200 mg/ml streptomycin and treated with 20 ml of HBSS containing 1 mg/ml collagenase (Sigma) for 30 min at 37°C. Following collagenase treatment, the SB202190 nmr supernatant was discarded and the tissue fragments were digested three

times with 0.25% trypsin and 3 mM EDTA in 20 ml of HBSS for 10 min at 37°C. The cells suspension was supplemented with 10% of heat-inactivated fetal bovine serum (FBS) to stop the activity of trypsin. AZD1152 To remove undigested tissue clumps, the cell suspension was passed through cell strainers (100-micro pores). To separate epithelial cells, which quickly formed

cell aggregates, from erythrocytes, platelets, and other immune cells, the cell suspension was centrifuged at 50 × g for 5 min. Following centrifugation, supernatant containing fibroblasts, erythrocytes, and immune cells, was discarded and the loose pellet containing epithelial cells and cell sheets was resuspended in 20 ml of HBSS. After three low-speed centrifugations, the cell pellet was resuspended in minimal essential CHIR98014 medium (MEM, ATCC) supplemented with 10% FBS, 2% heat-inactivated chicken serum (CS), insulin (0.12 U/ml), and estradiol (50 nM). The COEC cells were incubated in Petri dishes for 2 h at 39°C in 5% CO2 to allow fibroblast cells to attach. Following incubation, epithelial cells were collected by Atezolizumab price gentle pipetting and subsequent centrifugation at 125 × g for 10 min. The pelleted epithelial cells were resuspended in fresh MEM medium and seeded into 48-well tissue culture plates at a density of approximately 8 × 104 cells per well and incubated for 24 h to 48 h at 39°C in 5% CO2 until infection took place. Immunohistochemistry COEC cultures were incubated with monoclonal anti-pan cytokeratin

mouse Ab (epithelial cell marker) for 2 h at 37°C, washed three times, then incubated with fluorescein isothiocyanate (FITC) anti-mouse IgG for 1 h at 37°C. Staining of cytoskeleton of COEC was viewed with an Olympus IX81 FA scope. Cultures with more than 80% of cytokeratin-positive cells were used in subsequent infections. Thus, the COEC preparations consisted of more than 80% epithelial cells, less than 20% fibroblast, and possibly residual amount of immune cells. Infection of cell culture Infections were conducted using the gentamicin protection method as described previously [25]. Prior to inoculation, cell cultures were washed 3 times with pre-warmed Hanks’ Balanced Salt Solution (HBSS) without antibiotics. For each bacterial strain/time point combination, 500 μl of bacterial suspension containing approximately 16 to 24 × 105 CFU was added into each of the six wells to reach a multiplicity of infection (MOI) of 20:1 to 30:1 (bacteria:cells).

7A and lane 6 in Fig. 7B), as described above. These may be partially due to occurrence of IVS within the 16S rRNA genes from these isolates and fragmentation of the primary 16S rRNA transcripts among these isolates. However, we have not clarified the nature of the 16S rRNA genes from these isolates, yet. Therefore, sequencing and alignment analyses of the complete 16S rRNA genes from these isolates are VRT752271 cell line needed to identify the nature of the rRNA from these two Campylobacter species. Research to examine this is now in progress. Conclusions

Consequently, in 267 isolates of 269 Campylobacter isolates of the nine species (n = 56 C. jejuni; n = 11 C. coli; n = 33 C. fetus: n = 65 C. lari; n = 43 C. upsaliensis;

n = 30 C. hyointestinalis; CYT387 nmr n = 14 C. sputorum; n = 10 C. concisus; n = 7 C. curvus) examined, the absence of IVSs was identified in helix 25 region within 23S rRNA genes. Thus, IVS is extremely rare in the helix 25 region within the 23S rRNA genes from the Campylobacter organisms. The occurrence of IVSs with the two typical Campylobacter species, were shown in helix 45 region at a high percentage (54% for C. jejeuni n = 56; 45% for C. coli n = 11). We also identified the majority WZB117 in vitro (62/83) of isolates from the three Campylobacter species of C. fetus, C. upsaliensis and C. curvus to carry IVSs in helix 45. However, in a total of 54 isolates of the three species of C. hyointestinalis (n = 30), C. sputorum (n = 14) and C. concisus (n = 10), no IVSs were identified in the region. Thus, check details in conclusion,

no IVSs were identified in 105 isolates of three Campylobacter species (C. hyointestinalis, C. concisus and C. lari) both in the 25 and 45 helix regions. In addition, intact 23S rRNAs were identified in the purified RNA fractions in Campylobacter isolates containing no IVSs, and no 23S rRNA and fragmented other smaller RNA fragments were evident in the isolates containing IVSs. Methods Campylobacter isolates and genomic DNA preparation A total of 204 Campylobacter isolates [C. jejuni (n = 56); C. coli (n = 11); C. fetus (n = 33) C. upsaliensis (n = 43); C. hyointestinalis (n = 30); C. sputorum biovar sputorum (n = 4); biovar fecalis (n = 5); biovar paraureolyticus (n = 5); C. concisus (n = 10); C. curvus (n = 7)] were used in the present study (Table 2). Genomic DNA was prepared from Campylobacter cells by cethyltrimethyl ammonium bromide and proteinase K treatments, phenol-chloroform extraction and ethanol precipitation [23]. PCR amplification, cloning and sequencing We have already designed two PCR primer pairs, f-/r-Cl23h25, constructed to amplify helix 25 region and f-/r-Cl23h45, helix 45 region within the 23S rRNA gene sequences, based on the 23S rRNA gene sequence information from 12 UPTC isolates (DDBJ/EMBL/GenBank accsssion numbers, AB287301-AB287312), C. jejuni TGH9011 (Z29326) and C. coli VC167 (U09611) (Fig. 8) [22].

E. coli is among the most prevalent causes of hospital-acquired and community-acquired bacterial infections and their resistances to antimicrobial agents have become a serious concern for healthcare providers [5]. Phylogenetic analyses have classified E. coli into four main phylogenetic groups (A, B1, B2, and D). Commensal isolates belong mainly to A and B1 groups whereas virulent extra-intestinal pathogenic

E. coli (ExPEC) are essentially from the B2 and D groups [12, 13]. ExPEC harbor numerous virulence factors including α-hemolysin, cytotoxic necrotizing factor, adhesins and iron acquisition systems [12]. The spread of bla CTX-M-15 has been mainly associated with the dissemination of a particular clone of E. coli ST131 belonging to phylogenetic LGX818 purchase group B2 [14, 15]. Recently, an E. coli clone O25 ST131, producing CTX-M-15, with high virulence potential and belonging to the B2 group, has been reported and represent a

major public health problem [14, 15]. Many reports have documented the emergence of ESBL-producing Enterobacteriaceae[16–18]. In Antananarivo, ESBLs were first detected in 2005 from UTI in 9.7% of isolated Enterobacteriaceae[19]. In 2006, outbreaks of CTX-M-15 and SHV-2-producing K. pneumoniae isolates have been described in two pediatric units [20]. More recently, 21.3% of clinical isolates from patients in surgery and intensive care units [21] and 21.2% of intestinal carriage isolates from children hospitalized in a pediatric department of a large teaching find protocol Hospital [22] were ESBL-producers. For 49 Selonsertib in vivo multidrug-resistant Enterobacteriaceae isolates from Antananarivo, we characterized: i) the genes encoding the ESBLs; ii) the drug resistance genes associated with the ESBL genes; iii) gene cassettes present in the isolates; and iv) the plasmid incompatibility groups of the isolates. We also

determined the phylogenetic groups and virulence factors of the E. coli isolates. Methods Ethical clearance The study Flavopiridol (Alvocidib) protocols were approved by the National Ethics Committee of Madagascar. Written informed consents were obtained from all patients and at least one parent of each child before enrollment. Patients Between September 2006 and December 2007, a total of 909 non-duplicate bacterial isolates were obtained from 909 patients. 830 patients were recruited from several wards in four hospitals in Antananarivo, Madagascar (two national university teaching hospitals: Joseph Ravoahangy Andrianavalona Hospital and Befelatanana Hospital; a military hospital: Soavinandriana Hospital; and a pediatric hospital: Tsaralalana Hospital) and 79 patients referred to the Pasteur Institute Medical Laboratory in Antananarivo. Laboratory methods Various clinical specimens (including blood-culture, urine, pus, sputum and CSF) were collected and submitted for bacterial analysis at the Pasteur Institute Medical Laboratory in Antananarivo.

Phys Rev B 2009, 79:115409.CrossRef 39. Ding Y, Wang Y, Ni J, Shi L, Shi S, Tang W: First principles study of structural, vibrational and electronic properties of graphene-like, MX2 (M=Mo, Nb, W, Ta; X=S, Se, Te) monolayers. Physica B Condens Matter 2011,406(11):2254–2260.CrossRef 40. Ao Z, Li S, Jiang Q: Correlation of the applied

electrical field and CO adsorption/desorption behavior on Al-doped selleck inhibitor graphene. Solid State Commun 2010,150(13–14):680–683.CrossRef 41. Tang S, Cao Z: Adsorption of nitrogen oxides on graphene and graphene oxides: insights from density functional calculations. J Chem Phys 2011,134(4):044710.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions QY performed the first-principles calculations and drafted the manuscript. ZS and SC participated in the calculation part. JL conceived of the study and helped in writing of the manuscript. All

authors read and approved the final manuscript.”
“Background As superhard (hardness H ≥ 40 GPa) film material, nanocomposite films have been widely investigated in the past decades for use as wear-resistant coatings on tools and mechanical components [1, 2]. Among them, the pseudobinary TiN/SiN x is a representative film due to strong surface segregation of the constituent phases (TiN and SiN x have essentially no solid solubility). Especially, since hardness as high as 80 to 105 GPa was reported by Veprek et al. in 2000 [3], it has attracted much attention from the scientific community. So far the nanostructure and hardening mechanism have been widely

explained by nc-TiN/a-SiN x model proposed by Veprek Adriamycin et al. in 1995 [4], in which equiaxed TiN nanocrystallites (nc-TiN) were embedded in an amorphous SiN x (a-SiN x ) matrix. However, Cyclin-dependent kinase 3 this model is in dispute due to the lack of direct experimental evidence, which mainly reflects in two aspects. On one hand, whether TiN crystals are transformed from columnar crystals into equiaxed nanocrystallites is disputed, since there was no direct cross-sectional transmission electron microscopy (TEM) observation for the isotropic nature of the TiN grain. On the other hand, whether SiN x phase exists as amorphous state is also disputed, since Veprek et al. [4] suggested SiN x was amorphous because no obvious SiN x Bragg reflections in X-ray diffraction (XRD) patterns were found, which lacked direct observational evidence so far. Later, based on their high-resolution TEM (HRTEM) observations, Kong et al. [5] reported that TiN were columnar nanocrystals, rather than equiaxed nanocrystals, separated by crystallized SiN x interfacial phases. Hultman et al. [6] suggested that SiN x interfacial phase could be crystalline located around TiN nanocrystals according to their ab selleck initio calculations. However, they did not give direct experimental evidence. In addition, the cross-sectional TEM published by Zhang et al.

Chrysodontes (as subsect. Chrysodontini) within sect. Hygrophorus. Bon (1990) however, placed H. chrysodon in subg. Hygrophorus sect. Ligati (invalid). The yellow color and the glutinous pileus and stipe of sect. Chrysodontes differs from the dull colors and dry Emricasan basidiomata in sect. Camarophyllus, but the placement is supported by Larsson’s (2010) and our LSU analysis. Most authors did not classify H. inocybiformis (sect. Rimosi), but Fries (1874) placed it in subg. Camarophyllus, and Bon (1990), placed it in subg. Neocamarophyllus Bon [illeg.] sect. Neocamarophyllus Bon [illeg.] together with H. camarophyllus, H. calophyllus, and H. marzuolus. Although Bon’s (1990) group is most concordant with our molecular

phylogenies, his attempts to erect subgenus and sect. Neocamarophyllus were illegitimate because they lacked designated type species and Latin diagnoses. LY2090314 price As noted by Bas (1990), the citation by Arnolds (1990) as tribe Hygrophoreae (Kühner) Bas & Arnolds was incorrect in two respects: 1. tribe Hygrophoreae was published earlier than Kühner by P. Hennings (1898), and 2. only names below genus are recombined (Art. 6.7), so authors of higher Androgen Receptor phosphorylation taxa remain the same when they are transferred to another family. Bas (1990) and Arnolds (1990) treated tribe Hygrophoreae in the fam. Tricholomataceae rather than Hygrophoraceae. Hygrophorus [subgen. Camarophylli ] sect. Camarophylli

P. Bupivacaine Karst. [as Hygrophorus sect. Camarophyllus], Bidr. Känn. Finl. Nat. Folk. 25: 197 (1876). Type species Agaricus camarophyllus Alb. & Schwein. Consp. Fung. Lusat.: 177 (1805) : Fr. [Art. 22.6] [as H. caprinus (Scop.) Fr.], ≡ Hygrophorus camarophyllus (Alb. & Schwein. : Fr.) Dumée, Grandjean & L. Maire, Bull. Soc. mycol. Fr. 28: 292 (1912), [= Hygrophorus caprinus (Scop.) Fr. (1838), superfluous to a sanctioned name, nom. illeg., Art. 13.1]. Basidiomes dry; pileus grayish blue, grayish brown, buff brown, reddish brown bistre

or fuliginous; lamellae decurrent to deeply decurrent, white, sometimes with a grey or salmon-orange tinge; stipe grayish blue, grayish brown, buff brown, bistre or fuliginous; surface smooth or fibrillose. Lamellar trama divergent. Phylogenetic support Species in this clade are not represented in our LSU, ITS-LSU or Supermatrix analyses. Our ITS analysis places H. camarophyllus on a separate branch near the base of Hygrophorus, but without backbone support. Sect. Camarophylli is also basal in the four-gene analysis presented by E. Larsson (2010, unpublished data), comprising H. atramentosus, H. camarophyllus, H. calophyllus, H. capriolarius, and H. marzuolus, but without backbone support. Species included Type species: Hygrophorus camarophyllus. Additional phylogenetically supported species are H. atramentosus (Alb. & Schwein.) H. Haas & R. Haller Aar., H. calophyllus P. Karst., H. capreolarius Kalchbr. and H. marzuolus (Fr.) Bres.

sakazakii and C. malonaticus strains (Table 1 and Additional file 1). Reaction conditions for all the primers were as follows: initial denaturation at 94°C for 2 min; 30 cycles of denaturation at 94°C for 1 min, WZB117 purchase primer annealing at 58°C for 1 min, extension at 72°C for 2 min; followed by a final extension step of 72°C for 5 min. Each 50 μl amplification reaction mixture comprised ~10 ng chromosomal DNA, 10 μl Q solution (Qiagen, Crawley, UK), 20 pmol forward and reverse primer, 1× PCR buffer (Qiagen) containing 1.5 mM MgCl2, 0.8 mM deoxynucleotide triphosphates and 1.25 U Taq (Qiagen). The amplification product was then purified using MinElute UF plates (Qiagen) following the manufacturer’s protocol before being used in a sequencing

SHP099 mw reaction. Multilocus sequence analysis Using the nested sequencing primers, nucleotide sequences were determined at least once on each DNA strand with BigDye Terminator Ready Reaction Mix v3.1 (PE GDC-0449 research buy Biosystems, Foster City, US) under standard sequencing conditions according to the manufacturer’s protocol. Unincorporated dye terminators were removed by precipitation with 95% alcohol. The reaction products were separated and detected on an ABI PRISM genetic analyser 3100 (PE Biosystems) using a standard sequencing module with a Performance Optimised Polymer and 5 cm array. The sequences from both strands of a given locus of the same isolate were aligned, trimmed to the desired length and edited using SeqMan II (DNA

Star software, Madison, US). Allele and Sequence Type designation Arbitrary allelic numbers were assigned to each unique allele for a given locus. After sequencing and assigning allele types to all seven loci each isolate was then designated by a combination of seven numbers called an allelic profile that represented a sequence type (ST) for that particular isolate (eg.

ST1). A novel sequence type (ST) designation was given to each isolate with a unique allelic profile while subsequent isolates with an identical allelic profile were assigned the same ST identifier and considered to be isogenic strains as they were indistinguishable PD184352 (CI-1040) at all seven loci. All alleles within the MLST scheme were in frame, to aid with analysis. Linkage analysis Linkage analysis was carried out by using the index of association (I A ), as defined previously [37]. We examined whether alleles were randomly associated, that is, at linkage equilibrium, indicating a freely recombining population, or non-randomly associated, that is, at linkage disequilibrium, implying a clonal population structure. If there is linkage equilibrium, i.e., a random association between alleles of different loci, I A = 0. If I A is significantly different from 0, it indicates that recombination has been rare or absent and that the population has a clonal structure [34]. Acknowledgements The authors thank Nottingham Trent University, Micropathology Ltd and the Medical Research Fund for the funding of this study.

Like other ribozymes, HDV ribozyme has this property. So it may have a potential application in gene therapy in which an engineered ribozyme is directed to inhibit gene expression by targeting a specific PLX 4720 mRNA molecule. As hepatocellular carcinoma is often associated with the infection of HBV and HDV, The

facts that HDV ribozyme derived from HDV and that pathogen naturally infects and replicates in hepatocytes suggest that it can be used to control gene expression in human cells. The HDV ribozyme is active in vitro in the absence of any proteins, it is the only known example of a catalytic RNA associated with an animal virus. there are no known homologues of HDV ribozymes, and sequence variation of the HDV ribozymes in clinical isolates is minimal. FDA-approved Drug Library cell line Then we imagine whether HDV ribozyme can be used to inhibit hepatocellular carcinoma. In the present study we designed a HDV ribozyme against RNA component of human telomerase in hepatocellular carcinoma cell lines,

as well as in normal hepatocytes and other cancers, then examined the function of the HDV ribozyme and the effects of developing the HDV ribozyme as a tool of cancer gene therapy Methods The bel7402, HCT116 cells were given by Department of molecular Biology, Shandong University, DNA of HDV ribozyme was BMS345541 chemical structure synthesized by Shanghai Biosun Sci&Tech. Co. LTD. Recombinant plasmid pBBS212 containing hTR gene was provided by Geron Company. Design and synthesis of HDV ribozyme It was demonstrated that antigenomic ribozyme of HDV (g.RZ 1/84) is composed of 84 nucleotides[9]. It composed four stems (P1-P4), two loops and three junctions. As seen in Figure 1. Figure 1 Structure of antigenomic ribozyme of HDV (g.RZ Erythromycin 1/84). gRZ.1/84 can cleave 8-13 nt substrate by inter-molecular cleavage [10], the substrate must integrate with P1 stem of HDV ribozyme through base-pairing before cleavage, only 7 nt base pairing are needed, then the cleavage can occur. In P1 stem

G.U wobbling pair is essential for the activity of gRZ.1/84 and cannot be changed. The other 6 nucleotides can be changed, but the change must keep Waston-Crick pairing to substrate [11–13]. P4 stem isnot essential and can be deleted for easier access of ribozyme to substrate [14]. The activities of modified ribozyme do not decrease, but sometimes increase [15, 16]. We chose 12-84 nt of g.RZ 1/84, deleted 16 nt from P4 stem, and changed 6 nt of P1 stem from CCGACC to GGUUGA, only keeping G.U wobbling pair, to meet the need of cleavage of telomerase. We called the new ribozyme g. RZ57. The double-sranded DNA of g. RZ57 was synthesized with ApaΙ and HindIII protruding ends. Their sequences are as follows: 5′ AGCTT GGGAC CACCA CCACG CGGAC GCAAG AAGGG CAAGC GGCAA CGCAA GGCAA AGGGACCC CCC 3′ and 5′ A CCCTG GTGGT GGTGC GCCTG GCTGG TCCCG TTCGC CGTTG CGTTC CGTTT CCCTG GG GGG 3′. The predicted secondary structure of g. RZ57 are seen in Figure 2.